>>THANK YOU
FOR JOINING TODAY.
I AM DR. ANDREA DEPUTY DIRECTOR
OF THE NIH BRAIN INITIATIVE, AND
I'M VERY HAPPY TO BE HERE TODAY
TO INTRODUCE THE SESSION ALONG
WITH MY COLLEAGUE, DR. JAY
CHURCHILL.
IT'S PARTICULARLY APPROPRIATE
THAT WE ARE SHARING THIS
RESPONSIBILITY TODAY BECAUSE JAY
AND I WERE TRAINEES TOGETHER.
WE WERE POSTDOCS IN THE SAME LAB
MANY YEARS AGO, TOO MANY THAT I
WON'T MENTION RIGHT NOW.
AT ANY RATE, I WOULD LIKE TO
WELCOME YOU ALL TO THIS YEAR'S
TRAINEE HIGHLIGHT AWARDS TALKS,
TRAINEES ARE DEFINED AS HIGH
SCHOOL, UNDERGRADUATE, GRADUATE
AND MEDICAL SCHOOL STUDENTS AS
WELL AS POSTDOCTORAL FELLOWS AND
MEDICAL RESIDENTS.
THIS YEAR WE RECEIVED OVER 120
HIGHLIGHTED THE TRAINEES'
SPECIFIC CONTRIBUTIONS TO THEIR
MENTOR'S BROADER BRAIN AWARDS.
THE REVIEW TEAM WORKED REALLY
HARD ALONG WITH THE MEETING
PLANNING COMMITTEE AND SELECTED
THE TOP 30 FINALISTS TO PRESENT
FLASH TALKS TODAY DURING THIS
YEAR'S MEETING PROGRAM.
ALONG WITH AN INTERACTIVE
VIRTUAL POSTER.
AN ADDITIONAL 30 HONORABLE
MENTION RECIPIENTS WERE SELECTED
TO DISPLAY THEIR RESEARCH IN
THIS YEAR'S POSTER GALLERY AS
WELL.
>> AS ANDREA SAID, WELCOME TO
THE SESSION.
WE'RE REALLY, REALLY HAPPY TO BE
HERE AND WE ARE POSITIVE THAT
YOU'RE GOING TO HEAR SOME REALLY
FANTASTIC SCIENCE FROM PROMISING
INVESTIGATORS.
SO AS ANDREA MENTIONED, MY NAME
IS JAY CHURCHILL AND I SERVE AS
THE CO-LEAD FOR THE TRAINING,
INCLUSION AND EQUITY TEAM FOR
THE BRAIN INITIATIVE AT THE NIH.
AND REALLY IN A BIT OF A
SELF-SERVING POSITION, I'LL SAY
IF YOU HAVE INTEREST IN FUNDING
OPPORTUNITIES, PARTICULARLY AT
YOUR CAREER STAGE, PLEASE,
PLEASE, PLEASE REACH OUT TO US.
WE ARE PUBLIC SERVANTS, WE ARE
HERE TO HELP YOU, BUT WE CAN'T
HELP, WE CAN'T PROVIDE THAT
GUIDANCE IF YOU DON'T REACH OUT
TO US FIRST AND I'LL PUT MY
EMAIL INFORMATION IN THE CHAT
BOX FOR PEOPLE AND PLEASE SPREAD
THIS AROUND.
SO FOR TODAY, AWARD RECIPIENTS
WERE SELECTED THROUGH A PROCESS
BASED ON MULTIPLE FACTORS.
SOME OF THESE CRITERIA INCLUDE
THINGS SUCH AS THE SCIENTIFIC
FOCUS OF THE WORK, WITH
PRIORITIZATION FOR NULLIFYING
THESE OR FUTURE RESEARCH
DIRECTIONS IN AREAS OF HIGH
PRIORITY FOR THE NIH BRAIN
INITIATIVE.
ANOTHER FACTOR INCLUDES HOW THE
TRAINEES' RESEARCH CONTRIBUTES
TO THE LARGER BRAIN-RELATED
RESEARCH EFFORT WITH
CONSIDERATION AS TO WHETHER IT
INTEGRATES OR COMPLEMENTS
ONGOING EFFORTS WITHIN THE LAB,
AND IN PARTICULAR, PROJECTS
SUPPORTED DIRECTLY THROUGH THE
BRAIN INITIATIVE.
ANOTHER FACTOR IS IF THE
APPROACH PROVIDES GREAT EXAMPLES
OF INCLUSION STRATEGIES THAT
ADVANCE THE SCIENTIFIC AND
TECHNICAL MERIT OF THE RESEARCH
THROUGH ENHANCED INCLUSIVITY.
THERE ARE MANY REALLY GREAT
PROJECTS SUBMIT BID TRAINEES
ACROSS THE RESEARCH TRAINING
PIPELINE, UNFORTUNATELY WE CAN
ONLY SELECT A SMALL NUMBER THAT
YOU'LL HEAR ABOUT TODAY.
WHAT YOU'LL HEAR SHORTLY ARE
RESEARCH HIGHLIGHTS FROM 15 OF
THESE TRAINEES.
YOU SEE A LOT OF THESE REALLY
GREAT SMILING FACES IN FRONT OF
YOU NOW.
THERE ARE 15 MORE OF THEM BEING
PRESENTED SIMULTANEOUSLY IN
ANOTHER PANEL.
WE ARE ONLY -- WE GOT THE BEST
15 HERE, THOUGH.
DON'T TELL ANYBODY ELSE THAT.
WHAT WE ASK YOU AS FAR AS A
MATTER OF PROCESS IS THAT IF YOU
HAVE QUESTIONS FOR THESE PEOPLE
PLEASE USE THE QUESTION AND
ANSWER BOX.
AND IF YOU HAVE A SPECIFIC
QUESTION FOR ONE OF THESE,
PLEASE MENTION THEIR NAME AS
WELL SO WE CAN DIRECT THE
QUESTION TO THEM.
ANDREA AND I WILL ATTEMPT TO
RELAY THESE QUESTIONS TO THE
PRESENTERS AFTER WE HAVE
FINISHED HEARING FROM ALL 15 OF
THESE REALLY GREAT
INVESTIGATORS.
SO AGAIN, JOIN US IN WELCOMING
OUR TRAINEE HIGHLIGHT AWARD
PRESENTERS.
>> HELLO, MY NAME IS DESIREE
JOHNSON, RESEARCH ASSISTANT AT
THE UNIVERSITY OF CALIFORNIA
SAN DIEGO.
OUR BRAIN INITIATIVE FUNDED
RESEARCH AIMS TO IMPROVE DRUG
TARGETING IN THE NERVOUS SYSTEM
USING LIGHT.
WE DEVELOP PHOTO ACTIVATABLE OR
CAGED MOLECULES ADMINISTERED IN
AN BE INACTIVE CAGED FORM AND
LATER ACTIVATED WITH LIGHT.
PHOTO ACTIVATION OFFERS SPATIAL
AND TEMPORAL PRECISION, AS WELL
AS A ROBUST STIMULUS RESPONSE
RELATIONSHIP WHICH CREATES
OPPORTUNITIES TO QUANTIFY DRUG
RESPONSES IN NEW WAYS.
THIS IS NOT ONLY USEFUL FOR
MECHANISTIC INVESTIGATIONS INTO
DRUG ACTION BUT ALSO HAS
POTENTIAL APPLICATIONS IN
BIOMEDICINE.
BY RESTRICTING ILLUMINATION TO A
SPECIFIC REGION OF THE BODY OR
NERVOUS SYSTEM, PHOTO ACTIVATION
OF OPIOID DRUGS COULD YIELD
TARGET PAIN RELIEF WHILE
MINIMIZING SIDE EFFECTS SUCH AS
RESPIRATORY DEPRESSION AND
ADDICTION.
PREVIOUSLY, CAGED COMPOUNDS HAVE
BEEN PRIMARILY USED IN EX VIVO
PREPARATION SUCH AS BRAIN
SLICES, BUT WE ARE MOVING PHOTO
PHARMACOLOGY IN VIVO TO STUDY
HOW DRUGS AND NEUROMODULATORS
INFLUENCE ANIMAL BEHAVIOR.
YOU MAY HAVE ALREADY SEEN MY
OTHER POSTER DESCRIBING A CAGED
VAIR YABT OF
VARIANT OF THE SMALL MOLECULE
DRUG OXYMORPHONE THROUGH A
FIBEROPTIC CABLE IMPRINTED IN
THE MOUSE BRAIN.
I DESCRIBE OUR VALIDATION OF A
CAGE DERIVATIVE OF THE MU OPIOID
RECEPTOR AGONIST THAT CAN BE
INFUSED INTO THE RODENT BRAIN
THROUGH A CANNULA.
AND THEN PHOTO ACTIVATED WITH UV
LIGHT ACCORDING TO THE REACTION
SHOWN HERE.
WE THINK THAT THIS TOOL WILL BE
USEFUL TO STUDY HOW
OPIOID-SENSITIVE CIRCUITS
INFLUENCE BEHAVIOR.
AFTER IN VITRO AND EX VIVO
VALIDATION IN BRAIN SLICES TO
VALIDATE IN VIVO, I IMPLANTED
CANNULAS JUST ABOVE THE VENTRAL
TEGMENTAL AREA WHERE OPIOID
SIGNALING ACTIVATES DOPAMINE
NEURONS TO DRIVE BEHAVIORAL
REINFORCEMENT AND ENHANCE
LOCOMOTION.
I LOCALLY ADMINISTERED AND
SUBSEQUENTLY PHOTO ACTIVATED AT
THE SAME TIME 15 MINUTES LATER
USING A 375 NANOMETER LASER
WHILE MICE WERE EXPLORING IN
OPEN FIELDS.
DELIVERY OF LIGHT CAUSED A MERE
INSTANTANEOUS INCREASE IN
LOCOMOTION, REPRESENTED HERE AS
VELOCITY THAT WAS BLOCKED BY THE
OPIOID RECEPTOR ANTAGONIST
NALOXONE.
QUANTIFIED ON THE RIGHT, YOU CAN
SEE NALOXONE RETURN DISTANCE
TRAVELED TO LEVELS SIMILAR TO
VEHICLE AND NO LIGHT CONDITIONS.
TAKEN TOGETHER, OUR DATA
DEMONSTRATE THAT IN VIVO
UNCAGING IS A VIABLE APPROACH TO
MANIPULATING NEURAL CIRCUIT AND
BEHAVIOR WITH TEMPORAL PRECISION
ALL IN A TIME SCALE OF SECONDS.
WE ARE APPLYING THIS APPROACH TO
VARIOUS NEUROPEPTIDE SYSTEMS AND
TO OTHER INTERESTING THERAPEUTIC
DRUGS SUCH AS KETAMINE.
WE ARE ALWAYS INTERESTED IN
COLLABORATION SO IF PHOTO
PHARMACOLOGY COULD ADVANCE YOUR
RESEARCH PROGRAM, PLEASE REACH
OUT.
>> HELLO, EVERYONE.
I'M A POSTDOCTORAL SCHOLAR AT
UNIVERSITY OF CALIFORNIA
BERKELEY.
PERCEPTION IS WIDELY
MISCONCEPTUALIZED AS FAITHFUL
REPRESENTATION OF SENSORY
INFORMATION.
BUT IF THIS WERE THE CASE, WE
WOULD NOT BE SUSCEPTIBLE TO
ILLUSIONS.
ILLUSIONS ARISE WHEN THE BRAIN'S
INFERRED PERCEPTUAL OBJECT DOES
NOT MATCH THE SENSORY EVIDENCE.
HERE, WE UTILIZE CONTOURS TO
DETERMINE THE PERCEPTUAL
INFERENCE IN THE MOUSE VISUAL
CORTEX.
WE USE TWO PHOTO SCOPE TO
MEASURE VISUAL RESPONSES FROM
THOUSANDS OF NEURONS ACROSS SIX
SPATIAL AREAS SIMULTANEOUSLY.
WE STARTED BY COMPARING THE
ACTIVITY EVOKED BY ILLUSORY --
BY ROTATED CONTROLS.
THE PAIR OF ROTATED CONTROL
IMAGES INDICATED RC1 AND RC2 IN
THE FIGURE WAS DESIGNED TO MATCH
THE SUM OF PARTS IN THE PAIR OF
ILLUSORY CONTOUR IMAGES, IC1 AND
IC2.
THEREFORE, IF SENSORY
REPRESENTATION IS FAITHFUL, THE
SUM OF ACTIVITY EVOKED BY
ILLUSORY CONTOURS WOULD BE THE
SAME AS THE SUM OF ACTIVITY
EVOKED BY ROTATED CONTROLS.
TO OUR INITIAL SURPRISE,
HOWEVER, WE DID NOT SEE
POPULATION AVERAGE DIFFERENCES
IN ANY OF THE NEOCORTICAL VISUAL
AREAS BETWEEN THE ACTIVITY
EVOKED BY ILLUSORY CONTOURS
VERSUS ROTATED CONTROLS.
HOWEVER, THIS RESULT DOES NOT
INDICATE THAT NONE OF THESE
VISUAL AREAS COMPUTE PERCEPTUALR
INFERENCE.
THAT IS BECAUSE POPULATION
AVERAGE DIFFERENCE IS A
SUFFICIENT BUT NOT A NECESSARY
INDICATOR OF PERCEPTUAL
INFERENCE.
AS SUCH, WE TOOK AN ALTERNATIVE
APPROACH AND USED MACHINE
LEARNING DECODING TO ASSESS
NEURAL ACTIVITY PATTERNS.
SPECIFICALLY, WE USE THE SUPPORT
VECTOR MACHINE DECODER WITH A
LINEAR COLONEL.
KERNEL.
ACTIVELY DEPICTED FROM VISUAL
RESPONSES.
NOW TO ASSESS WHETHER THE
ILLUSORY CONTOUR WAS REPRESENTED
IN EACH VISUAL AREA, WE DESIGNED
A REAL EDGE STIMULUS THAT
INCLUDES A T SHAPE DENOTED
RET1 AND RE T2 IN THE FIGURE.
RET1 STIMULUS HAS EQUAL OVERLAP
WITH IC1 AND RC1.
THEREFORE, RET1 ACTIVITY WILL BE
EQUALLY LIKELY TO BE DECOATED S
IC1 OR RC1, IF AND ONLY IF
REPRESENTATION WAS FAITHFUL.
USING THIS APPROACH, WE FOUND
THAT THE INFERRED REPRESENTATION
OF THE ILLUSORY CONTOUR IS
PRESENT IN LAYER 23 OF V1, RM
AND RL, INDICATING FUNCTIONAL
SPECIALIZATION OF HIGHER VISUAL
AREAS.
ON THE OTHER HAND,
REPRESENTATION IN LAYER FOUR OF
V1 WAS FAITHFUL, SUGGESTING THAT
PERCEPTUAL INFERENCE EMERGES
ALONG THE VISUAL HIERARCHY.
ONGOING EXPERIMENTS AIMED TO
DETERMINE THE RECURRENT
EXCITATORY MECHANISMS BY WHICH
ILLUSORY CONTOUR REPRESENTATION
ARISES.
WE USE OPT GENETICS TO
SELECTIVELY STIMULATE NEURONS OF
INTEREST.
WHEN WE STIMULATED NEURONS THAT
SELECTIVELY RESPOND TO THE
EMERGENT PROPERTIES OF THE
ILLUSORY CONTOUR, IT RECREATED
THE NEURAL ACTIVITY PATTERN
EVOKED BY THE ILLUSORY CONTOUR
IMAGE.
IN OTHER WORDS, HOLE GRAPHIC
STIMULATION OF I ILLUSORY
CONTOURING DECODERS -- IN LAYER
23 OF V1 WAS SUFFICIENT TO DRIVE
THE NETWORK TO PATTERN
COMPLETION.
IF YOU'RE INTERESTED IN DETAILS,
PLEASE VISIT MY POSTER, NUMBER
825.
THANK YOU VERY MUCH FOR YOUR
ATTENTION.
>> HI, EVERYONE.
I'M A THIRD YEAR PH.D. STUDENT
AT MIT AND I STUDIED THE
ORGANIZATION OF THE SYNAPTIC
MOLECULAR NETWORK.
THIS IS A COMPLEX SYSTEM OF
HUNDREDS OF PROTEIN, EVEN MORE
INTERACTIONS COMPARTMENTALIZED
TO THE SYNAPTIC SITES.
THE SYSTEM IS INTERESTING
BECAUSE IT IS ESSENTIALLY THE
HARDWARE OF WHICH RUNS THE
COMPLEX COMPUTATION OF EACH
INDIVIDUAL SYNAPSE.
RESPONSE TO ACTIVITY CHEMICAL
AND GENETIC HISTORIES LEADING TO
PLASTICITY AND META PLASTICITY.
I STUDIED THE NETWORK -- I WANT
TO FIRST MEASURE THE
DISTRIBUTION OF SYNAPSES IN HIGH
DIMENSIONAL COMPOSITION SPACE
AND TWO, FIGURE OUT THE RULES
THAT GOVERN THIS DISTRIBUTION
AND THE PATHS THAT SYNAPSES TAKE
IN THAT SPACE.
TO ACHIEVE THE FORMER, I NEED A
WAY TO MEASURE MULTIPLE PROTEINS
IN MANY INDIVIDUAL SYNAPSES.
I DO THAT USING PRISSMM.
A HIGH THROUGHPUT QUANTITATIVE
MULTIPLEXED IMAGING TECHNIQUE
DEVELOPED IN THE LAB THAT RELIES
ON FLUORESCENT PROBE EXCHANGE ON
BARCODED ANTIBODIES.
THERE IS ALSO SUCH AN EXPERIMENT
SEEN IN THE TOP RIGHT ARE
SEQUENTIAL IMAGES OF THE SAME
NEURONAL CULTURE, EACH TIME
LIGHTING UP A DIFFERENT PROTEIN.
I CAN AUTOMATICALLY SEGMENT
INDIVIDUAL SYNAPTIC -- USING
CELL PROFILER AND QUANTIFY THEM
ACROSS CHANNELS, GETTING
MULTI-PROTEIN MEASUREMENTS FOR
MILLIONS OF INDIVIDUAL SYNAPSES.
I THEN USE BAYESIAN NETWORK
EVIDENCE FROM THIS DISTRIBUTION.
BAYESIAN NETWORK INFOR INSTANCE
ALLOWS ONE TO FACTOR THIS HIGH
DIMENSIONAL DATA INTO SINGLE
VARIABLE CONDITION OF THE
DISTRIBUTIONS SUCH AS THE ONE ON
THE BOTTOM RIGHT.
FROM ONE TO ANOTHER IMPLIES THAT
THE DISTRIBUTION OF THE CHANNEL
DEPENDS ON THE LEVELS OF THE
PARENT.
MORE INTERESTINGLY, LACK OF AN
EDGE INDICATES THAT EVEN THOUGH
TWO NODES MAY BE TIGHTLY
CORRELATED, ANY CONNECTION
BETWEEN THEM IS EXPLAINED BY
OTHER VARIABLES.
THE RESULTING MODEL GIVES --
PREDICTIONS ABOUT DOWNSTREAM
EFFECTS OF DETECTING CERTAIN
NODES.
FOR EXAMPLE, THE ACTIN 3 CHAIN
PREDICTS THAT FIRST DIRECT
PRESERVATION OF SHANK 3 SHOULD
AFFECT ONLY SHANK 3.
PSD95 SHOULD AFFECT BOTH
PSD95 AND SHANK 3 AND ACTIN
SHOULD AFFECT ALL THREE.
IN ADDITION, SINCE THE EFFECT ON
SHANK 3 SHOULD BE MEDIATED
MOSTLY BY CHANGING PSD95 LEVELS,
WHEN THE CONTROL FOR THAT, THE
EFFECT OF CHANGING ACTIVE ON
SHANK 3 SHOULD WEAKEN.
THIS IS EXACTLY WHAT I OBSERVED
WHEN TESTING THESE
PRESERVATIONS.
FINALLY, IN COLLABORATION
WITH -- PSYCHIATRIC RESEARCH AT
THE BROAD INSTITUTE, HOW THE
MOLECULAR SYSTEM AS A WHOLE IS
AFFECTED BY PERTURBING DIFFERENT
GENES ASSOCIATED WITH AUTISM AND
SCHIZOPHRENIA.
WE OBSERVE ALTHOUGH KNOCK DOWN
OF DIFFERENT GENES HAVE VARYING
EFFECTS ON THE SYNAPTIC PROTEINS
THEMSELVES, SOME NETWORK EDGES
WERE CONSISTENTLY STRENGTHENED
OR WEAKENED BUT ALL TREATMENTS
AS SHOWN IN COLORS IN THE GRAPH
ON THE RIGHT.
THESE PHENOTYPES MAY PROVIDE THE
CLUE -- BY WHICH DIFFERENT
AUTISM AND
SCHIZOPHRENIA-ASSOCIATED
MUTATIONS GIVE RISE TO SIMILAR
SYMPTOMS.
I AM CURRENTLY EXPANDING THIS
COMBINED EXPERIMENTAL AND
COMPUTATIONAL METHODOLOGY IN
SEVERAL DIRECTIONS.
COMBINING WITH IMAGING TO
INTEGRATE SYNAPTIC ACTIVITY
MEASUREMENTS INTO THIS PROTEIN
NETWORK.
DEVELOPING A FRAMEWORK FOR
THE -- USING PRISSMM, AND
APPLYING THE METHOD TO
CHARACTERIZE DOWNSTREAM
BIOCHEMICAL EFFECTS OF
PSYCHIATRIC DRUGS.
I HOPE THAT I PROMPTED AT LEAST
SOME OF YOU TO THINK ABOUT THIS
NEW KIND OF DATA MODALITY AND I
WOULD LOVE TO DISCUSS MORE IN
DEPTH AT MY POSTER.
THANK YOU.
>> HELLO, BAYLOR COLLEGE OF
MEDICINE, HOUSTON, TEXAS.
I'M VERY EXCITED TO DISCUSS WITH
YOU SOME NEW GENETICALLY ENCODED
GEVIs FOR LONG TERM PHOTON
IMAGES.
GEVI -- THROUGH CHANGES IN THEIR
FLUORESCENCE.
THIS CLASS OF SENSOR HAS MANY
ADVANTAGES OVER OTHER SENSING
TECHNIQUES SUCH AS
ELECTROPHYSIOLOGY, MICRO ARRAYS
AND CALCIUM IMAGING.
GEVIs ARE RELATIVELY
NONINVASIVE USING ONLY LIGHT TO
RECORD VOLTAGE DYNAMICS INSTEAD
OF ELECTRODES OR IMPLANTS.
THEY CAN ALSO BE CELL
TYPE-SPECIFIC WHEN EXPRESSED
USING A CELL-TYPE SPECIFIC
PROMOTER.
GEVIs CAN -- RESOLUTION AND
ARE EVEN ABLE TO DETECT
INDIVIDUAL ACTION POTENTIALS IN
FAST WAVE -- FINALLY, GEVIs
CAN HAVE HIGHER SPEACIAL
RESOLUTION BY FACILITATING --
THESE ARE JUST A FEW VAMS OF THE
AGAIN BEN FITS.
WHILE GREAT STRIDES HAVE BEEN
MADE TO IMPROVE THE AVAILABLE
TOOLBOX OF GEVIs, MANY
SCHEDULE WITH PERFORMANCE UNDER
PHOTON MICROSCOPY, A METHOD OF
CHOICE FOR DEEP TISSUE IMAGING.
MANY OF THESE SENSORS SUFFER
FROM PHOTO BLEACHING WHICH CAN
LIMIT THE LENGTH OF OF CONTINS
RECORDING DURING BEHAVIOR ASSAY
AND -- POTENTIAL DYNAMICS.
COUPLE THIS WITH THE RELATIVELY
DIM PHENOTYPE RESULTS AND
DIFFERENCES IN USING --
DIFFICULTIES IN USING TWO PHOTON
IMAGING IN DEEPER LAYERS OF THE
BRAIN FOR LONG PERIODS OF TIME.
THUS WE SET OUT TO DESIGN A
SENSOR THAT HAS IMPROVED OF
COURSE THROUGH PHOTON PHOTO
STABILITY, BRIGHTNESS AND
SENSITIVITY TO ACTION
POTENTIALS.
WE KNEW THAT LARGE LEAPS IN
SENSOR DESIGN COULD ONLY BE
ACHIEVED USING A HIGH THROUGH
BUT HOLISTIC CHARACTERIZATION
APPROACH.
SO WE DESIGNED A SCREENING
PLATFORM THAT ALLOWED US TO DO
MULTICHARACTER -- BRIGHTNESS,
PHOTO STABILITY AND RESPONSE TO
DEEP FLUOROIZATION FOR EACH
TESTED.
WE SCREENED OUR VARIANTS BASED
OFF OF THE ACES SCAFFOLD, A
VOLTAGE SENSITIVE DOMAIN
ATTACHED TO A CIRCLELY PER MUTED
EXTRACELLULAR GFP.
WE TARGETED STRUCTURALLY AND
EVOLUTIONARY RESIDUES INCLUDING
AMINO ACIDS TO INTERACT WITH THE
FLUORESCENT PROTEIN OR ARE
CRITICAL TO THE FUNCTION OF THE
VOLTAGE SENSING DOMAIN.
AFTER THOUSANDS OF VARIANTS
TESTED, WE'RE EXCITED TO SAY WE
IDENTIFIED A NEW GEVI NAMED
JELLYFISH DERIVED VOLTAGE
INDICATORS FOR TWO PHOTON
IMAGING.
OR JEDI-2P FOR SHORT, WHICH CAN
DETECT INDIVIDUAL ACTION
POTENTIALS FOR OVER 40 MINUTES
AND 270 MICRO METERS OF THE
SURFACE OF THE MOUSE BRAIN USING
TWO PHOTON MICROSCOPY.
AND IS SHOWN RIGHT HERE IN THE
SLIDE.
JEDI-2P HAS BEEN SUCCESSFULLY
TESTED IN ISOLATED MOUSE RETINA,
DROSOPHILA AND FISH.
THUS SHOWING GREAT -- JEDI-2P OF
COURSE IS NOT THE LAST JEDI, AND
WE'RE CONTINUING TO MAKE
ADVANCEMENTS.
SO IF YOU'RE INTERESTED IN
TRYING OUT JEDI-2P OR ANY OF OUR
NEWEST SENSORS, PLEASE CONTACT
US THROUGH EMAIL OR QR CODE.
OVER LONGER TIME SPANS AND
ADVANCE OUR UNDERSTANDING OF
NEURAL DATA PROCESSING IN HEALTH
AND DISEASE.
THANK YOU FOR YOUR TIME, AND IF
YOU HAVE ANY ADDITIONAL
QUESTIONS, PLEASE CONTACT TO
DISCUSS.
>> HELLO, EVERYONE.
I'M EXCITED TO TALK TO YOU ABOUT
MY RESEARCH, WHICH IS RESTORING
SENSATION AFTER LOWER LIMB
AMPUTATION USING SPINAL CORD
STIMULATION.
THE PEOPLE WITH LOWER LIMB
AMPUTATION HAVE PROCESSES WHICH
LACK SENSORY FEEDBACK.
NOW THE SENSORY FEEDBACK IS NOT
JUST IMPORTANT FOR SOMEONE TO
FEEL THEIR LEGS, BUT AT THE SAME
TIME, IT PLAYS A CRUCIAL ROLE IN
DIFFERENT LOWER LIMB FUNCTIONS
LIKE MAINTAINING BALANCE OR
GAIT.
SO WE WANT TO RESTORE THIS
MISSING SENSATION USING A
CLINICALLY APPROVED TECHNOLOGY
CALLED SPINAL CORD STIMULATION.
CURRENTLY THIS TECHNOLOGY IS
USED TO TREAT CHRONIC PAIN AND
WE WANT TO USE THE SAME
TECHNOLOGY TO SEE IF WE CAN
RESTORE MISSING 16 SENSATIONSN
THESE AMPUTEES.
SO WE HAVE IMPLANTED THREE
SUBJECTS SO FAR.
WE PLACED THESE FLEXIBLE LEADS
AS YOU CAN SEE IN THE FIGURE IN
THE LUMBAR REGION OF THE SPINAL
CORD, AND WE HAVE SEEN THAT
SUCCESSFULLY IN AMPUTEE, WE
COULD EVOKE SENSATIONS IN THE
MISSING LIMB, AND THESE EVOKED
SENSATIONS ALSO IMPROVE THE
LOWER LIMB FUNCTIONS.
I'M EXCITED TO SEE YOU ALL IN MY
POSTER.
THANK YOU FOR LISTENING
FLEURCHLT
>> HELLO, EVERYONE.
MY NAME IS NATALIE AND I WORK AS
A POSTDOC IN THE DEPARTMENT OF
BIOMEDICAL ENGINEERING AT BOSTON
UNIVERSITY.
IT'S A WIDELY USED TECHNIQUE IN
HUMANS TO STUDY BRAIN FUNCTION
IN HEALTH AND DISEASE.
HOWEVER, THIS TECHNIQUE DOES NOT
DIRECTLY MEASURE NEURONAL
ACTIVITY, BUT RATHER REFLECTS A
BALANCE BETWEEN OXYGEN DELIVERY
AND OXYGEN CONSUMPTION.
MY GOAL WAS TO ESTIMATE OXYGEN
CONSUMPTION OR CEREBRAL
METABOLIC RATE OF OXYGEN IN THE
BRAIN OF MICE WITH PHOTO
MICROSCOPY.
USING OXYGEN CONSUMPTION
TOGETHER WITH BLOOD FLOW AND
NEURONAL ACTIVITY,
MICROSCOPICALLY CAN BE USED TO
MODEL POLYMERIZED SIGNALS IN
HUMANS.
SO I USED OXYGEN ON AN OXYGEN
SENSOR OXYPHOR 2P -- THAT HAVE
BEEN USE FORD A LONG TIME,
OXYPHOR 2P ALLOWS ME TO MEASURE
TO THE HIGHEST SPATIAL
RESOLUTION.
I INJECTED THE DYE INTO THE I
HAVE SIPT OF AN
VICINITY IN THE MOUSE CORTEX.
THEN I FOLLOWED THE ARTERY DOWN
TO LAYER FOUR AND I CREATED --
AROUND THE ARTERY TO MEASURE HOW
OXYGEN DIFFUSES INTO THE TISSUE.
AT THIS POINT I MEASURE THE
PHOTON DECAY.
BECAUSE I EXCITE THE FOS PHOSS
IN DYE FIRST AND DEPENDENT ON
HOW MUCH OXYGEN IS PRESENT --
THEN I CONVERTED TO PO2 AND
CREATE A PO2 MAP ON TOP OF THE
VESSEL MAP.
AS YOU CAN SEE HERE IN THE
VESSEL MAP ON THE RIGHT SIDE,
THE OXYGEN IS HIGH INSIDE THE
VESSEL AND THEN IF YOU GO
TOWARDS THE PERIPHERY, THE
OXYGEN DROPS.
THIS DROP OF OXYGEN OR GRADE
YENT OF OXYGEN IS DEPENDENT ON
HOW MUCH OXYGEN IS CONSUMED.
THE DEEPER THE GREAT YENT, E
MORE OXYGEN IS CONSUMED.
AFTER I WENT THROUGH THE LAYERS
IN THE CORTEX, WE COULD SEE THAT
LAYER ONE IN BLUE HAS THE
STEEPEST GRADIENT, INDICATING
THE HIGHEST OXYGEN CONSUMPTION.
THIS IS IN CONTRAST TO THE
COMMON BELIEF IN THE FIELD THAT
OXYGEN CONSUMPTION IS HIGHEST IN
LAYER FOUR, BECAUSE IN LAYER
FOUR, WE HAVE THE HIGHEST
NEURONAL DENSITY AND ALSO
CAPILLARY DENSITY.
AND CYTOCHROME -- FOR OXIDATIVE
PHOSPHORYLATION LOCATED IN THE
MITOCHONDRIAL BRAIN, IT'S MORE
PREVALENT THEREFORE.
HOWEVER, THIS TECHNIQUE COULD
SHOW THIS IS NOT THE CASE IN THE
MOUSE CORTEX.
THEN WE THOUGHT IT MIGHT BE
UNDER STIMULATING CONDITIONS ON
THE AXIAL BRAIN STATES WHEN WE
NEED A HIGHER CONSUMPTION IN
LAYER FOUR.
HOWEVER, PRELIMINARY DATA SHOWS
AS WELL THAT THE OXYGEN
CONSUMPTION IS HIGHEST IN LAYER
ONE AND THEN DECREASES WITH
LAYER TWO THROUGH LAYER FOUR.
THE TECHNIQUE I PRESENT HERE
SHOWS A POWERFUL TOOL THAT CAN
BE USED IN HEALTH AND DISEASE
MODELS AND IT CAN BE USED TO
MODEL FMRI SIGNALS.
THANK YOU VERY MUCH FOR YOUR
ATTENTION.
>> HELLO, EVERYONE.
MY NAME IS CARL AND I'M EXCITED
TO TALK ABOUT MY RESEARCH HERE
AT UT SOUTHWESTERN MEDICAL
CENTER.
THE OVERALL GOAL OF MY PROJECT
IS TO UNDERSTAND THE CELL TYPES
THAT ARE ENGAGED IN THE BRAIN
CIRCUITS FOR VOCAL LEARNING AND
WHAT VOCAL LEARNING IS IS THE
ABILITY TO MODIFY VOCALIZATIONS
OR ACOUSTIC SOUNDS AND VOCAL
LEARNING IS A CRITICAL COMPONENT
OF HUMAN SPEECH, BUT IT'S
DIFFICULT TO STUDY IN CLASSICAL
MODEL ORGANISMS SUCH AS MICE AND
NON-HUMAN PRIMATES BECAUSE THEY
DO NOT EXHIBIT VOCAL LEARNING
BEHAVIOR.
SO IN OUR LAB, WE USED SONG
BIRDS WHICH LEARN THEIR SONG
VERY SIMILAR TO THE WAY HUMANS
LEARN LANGUAGE.
WE SPECIFICALLY USED ZEBRA
FINCHES BECAUSE THEIR VOCAL
LEARNING STAGES ARE MOST SIMILAR
TO HUMANS.
THEY BOTH NEED TO HEAR SPECIFIC
VOCALIZATIONS EARLY IN THEIR
DEVELOPMENT WHEN THEIR BRAIRNS
AND BRAINS ARE VERY
PLASTIC.
THE SENSORY LEARNING PERIOD OR
SENSORY LEARNING STAGE.
WE KNOW THAT THERE ARE
SIMILARITIES IN BEHAVIOR AND
DEVELOPMENT IN THEIR VOCAL
LEARNING BEHAVIOR WITH YOU O
IDENTIFIED -- FOR CONTROL OF
VOCALIZATION.
ALTHOUGH WE KNOW IN GENERAL THE
BRAIN REGION INVOLVED IN SPEECH
AND SOUND CONTROL, WE DON'T KNOW
WHAT IS OCCURRING AT THE
CELLULAR RESOLUTION AND HOW IT'S
CHANGING THROUGHOUT DEVELOPMENT
DURING THESE SENSITIVE PERIODS
WHEN THEY ARE LEARNING SONG OR
SPEECH.
SO HOW DO WE ANSWER THIS
QUESTION OF WHAT ARE THE CELL
TYPES ENGAGED DURING DIFFERENT
VOCAL LEARNING STAGES?
WELL, FOR THIS, WE DISSECTED A
SPECIFIC BRAIN REGION SONG BIRDS
NECESSARY BOTH FOR SONG LEARNING
AND PRODUCTION AND THAT IS
ANALOGOUS TO THE PREMOTOR CORTEX
FOR CONTROLLING VOCALIZATION IN
HUMANS.
WE CONDUCTED SINGLE RNA -- AT
DIFFERENT TIME POINTS AT 24 AND
45 DAYS POST HATCH AND WE
COMPARED TO ADULTS.
AND WE SPECIFICALLY FOCUSED ON
USING THIS DATA GENE EXPRESSION
DATA WHERE WE DID A PRINCIPAL
COMPONENT ANALYSIS AND REDUCTION
TO IDENTIFY MEAN CELL CLUSTERS
THAT WE CAN THEN LABEL USING THE
GEN EXPRESSION HERE.
WE CAN IDENTIFY THE MAJOR CELL
TYPES SUCH AS THE GABA AND OTHER
GLIAL -- THAT PROJECT EITHER TO
THE BASAL GANGLIA, AUDITORY
REGION OR MOTOR PATHWAY.
ONCE WE CAN IDENTIFY THESE VEL
TYPES, CELL
TYPES, WE CAN FOCUS ON HOW IT'S
CHANGING THROUGH THIS
DEVELOPMENT AND COMPARE TO
ADULTS WHICH IS SHOWN HERE, A
PERCENTAGE OF A SPECIFIC CELL
TYPE TO ALL OF THE PROJECTION
TYPES AND REMARKABLY WHAT WE
FOUND THAT HAS BEEN REPORTED
BEFORE BUT NOT WITH THAT TYPE OF
DATA IS THAT THE MOTOR
PROJECTING NEURONS IN ADULTS ARE
THE ONES THAT ARE THE LARGEST
PERCENTAGE IN ADULTS SHOWN HERE
IN THE BLACK BAR, BUT ARE NOT
REALLY AS HIGH AS JUVENILES.
BUT IF YOU CONTRAST THIS WITH
THE YOUNG BIRDS, YOU CAN SEE THE
AUDITORY PROJECTION NEURONS MAKE
UP THE MAJORITY OF THE
POPULATION WHILE IT REALLY
DECREASES DRASTICALLY IN ADULT
POPULATION.
SO THIS DEMONSTRATED THAT THE
DRAMATIC CHANGE IN CELL TYPE
COMPOSITION COULD MAKE A
POTENTIAL MECHANISM FOR HOW THIS
SOUND KROAL CONTROL REGION FS
WHEN THE BRAIN IS STILL MATURING
AND CAN CHANGE THROUGHOUT
DEVELOPMENT TO ACCOMMODATE
DIFFERENT METHODS OF LEARNING
THAT CAN APPLY NOT ONLY TO SONG
BIRD LEARNING BUT ALSO HUMAN
SPEECH LEARNING.
WITH THAT, I WANT TO THANK
EVERYONE FOR LISTENING TO MY
TALK AND I'LL BE HAPPY TO TAKE
ANY QUESTIONS HERE OR IN THE
POSTER SESSION.
THANK YOU.
>> HI, MY NAME IS MICHELLE
FALLA, GEORGETOWN UNIVERSITY,
BEGINNING MY SECOND YEAR IN THE
INTERDISCIPLINARY PROGRAM IN
NEUROSCIENCE.
I WORK IN DR. REBECCA EVANS' LAB
INVESTIGATING HOW THE PPN INACTS
WITH THE BASAL GANGLIA TO
MODULATE MOVEMENT.
PREVIOUS STUDIES HAVE
CONTRADICTORY OUTCOMES WHERE
SOMETIMES LOCOMOTION IS
INCREASED AND OTHER TIMES
LOCOMOTION IS DECREASED.
IN FACT, DEEP BRAIN STIMULATION
OF THE PPN IN A PARKINSONIAN RAT
MODEL RESULTED IN WORSENED GAIT
WHEREAS CAUDAL STIMULATION LED
TO IMPROVED GAIT.
FUNCTIONALLY DISTINCT
SUBPOPULATIONS WITHIN THE PPN
CAN HELP EXPLAIN VARIABLE
OUTCOMES IN THE PREVIOUS
LITERATURE, BUT ALSO VARIABLE
OUTCOMES OF PPN-TARGETED DEEP
BRAIN STIMULATION IN PARKINSON'S
DISEASE PATIENTS.
MY WORK FOCUSES ON THE
CHOLINERGIC PPN NEURONS WHOSE
DEGENERATION IN PARKINSON'S
DISEASE CORRELATES WITH GAIT
IMPAIRMENT IN BOTH PATIENTS AND
RODENT MODELS.
I'M USING A MOUSE MODEL TO
CHARACTERIZE THE PROPERTIES OF
THE PPN NEURONS WHICH WILL
FLORES RED UNDER GREEN LIGHT AS
YOU CAN SEE IN THIS FIRST IMAGE.
IT IS COMMON FOR YOUNG MICE TO
BE USED IN ELECTROPHYSIOLOGY
EXPERIMENTS STUDYING THE PPN.
BUT THIS IS NOT SUITABLE FOR
RESEARCH RELATED TO DISEASES
WHICH ARE MORE COMMON WITH AGE
LIKE PARKINSON'S DISEASE.
SO I USED WHOLE CELL PATCH CLAMP
TO RECORD NEURONAL ACTIVITY FROM
THE CHOLINERGIC PPN NEURONS AND
ANALYZED THE DATA BY SEX AND
AGE.
I FOUND SIGNIFICANT DIFFERENCES
IN THE INTRINSIC
ELECTROPHYSIOLOGICAL PROPERTIES
AND THE ROSTRAL CAUDAL
DISTRIBUTION OF CHOLINERGIC
ELECTROPHYSIOLOGICAL SUBTYPES IN
ADULT MICE COMPARED TO MICE
UNDER TWO MONTHS OF AGE.
WE ARE WORKING TO DETERMINE THE
FUNCTIONAL SIGNIFICANCE OF THESE
ROSTRAL CAUDAL DIFFERENCES.
SO WE WANTED TO DETERMINE IF THE
CAUDAL AND ROSTRAL PPN RECEIVE
DIFFERENTIAL INNERVATION FROM
THE INHIBITORY BASAL GANGLION
NUCLEI.
WE CAN OPT GENETICALLY STIMULATE
THE AXONS PRESENTING TO THE PPN
WHILE RECORDING THE INHIBITORY
INPUT TO THE CHOLINERGIC
NEURONS.
IN THIS IMAGE, WE CAN SEE HOW
THESE GREEN AXONAL PROJECTIONS
FROM THE GPE EXTEND OVER AND
THROUGH THE CAUDAL PPN.
THE DISTRIBUTION OF AXONAL
PROJECTIONS IN IMAGING WERE
SUPPORTED BY THE RECORDED
INPUTS.
THE SNR INNERVATES ABOUT 80% OF
THE NEURONS WITH RELATIVELY
EQUAL INNERVATION OF THE ROSS
STRAL AND CAUDAL PPN.
THE PPN RECEIVES MUCH LESS
INNERVATION FROM THE GPE WITH
ABOUT 40% OF THE PATCH NEURONS
RECEIVING INHIBITORY INPUT.
MOST OF THE NEURONS THAT WERE
CONNECTED TO THE GPE WERE
LOCATED IN THE CAUDAL PPN.
SUPPORTING WHAT WE WITH SEEN INN
THE IMAGING.
WE ARE IN THE PROCESS OF USING
TWO PHOTON IMAGING TO CLASSIFY
THE REGIONAL CONNECTIVITY
BETWEEN THESE NUCLEI IN THE PPN
ACROSS THE ROSTRAL CAUDAL ACCESS
AS WELL AS THE MEDIAL LATERAL
AXIS.
THE LAST THING I'LL SHARE IS
THAT WE STAINED FOR THE
EXTRACELLULAR MATRIX PROTEIN
NPNT BECAUSE IT IS ONLY REALLY
FOUND IN THE CORTEX AND OUR
REGION OF INTEREST, THE PPN.
WE WERE ABLE TO IDENTIFY AN
NPNT-NEGATIVE SUBGROUP WITHIN
THE CAUDAL PPN, SO WE PLAN TO
INVESTIGATE THE INTRINSIC AND
SYNAPTIC PROPERTIES WHICH MAY
DIFFERENTIATE THE CHOLINERGIC
NEURONS AND THE NPNT POSITIVE
CHOLINERGIC NEURONS.
THESE FINDINGS WILL HELP US
BETTER UNDERSTAND HOW THE PPN
RECEIVES REGION-SPECIFIC INPUTS
FROM THE BASAL BEGAN GANGLIAO
DETERMINE MODULATION ON MOTOR
OUTPUT.
THANK YOU FOR YOUR TIME, AND
PLEASE CHECK OUT POSTER NUMBER
808 FOR MORE INFORMATION.
>> HELLO, EVERYBODY.
I'M A POSTDOCTORAL SCHOLAR AT
THE CEDARS-SINAI MEDICAL CENTER.
TODAY I WOULD LIKE TO INTRODUCE
TO YOU MY RESEARCH IN THANK -- R
TO PERFORM A CERTAIN TASK.
SUCH DISTRACTIONS CAN COME FROM
THE EXTERNAL WORLD SUCH AS A
LOUD RAIN -- OR IT CAN COME FROM
INTERNAL SOURCES SUCH AS OUR OWN
HABITS OR INTRUSIVE THOUGHTS.
WHEN DISTRACTION INTERFERES WITH
OUR GOAL TO A MODERATE EXTENT,
WE EXPERIENCE CONFLICT.
WHEN DESTROYING ENOUGH TO KNOCK
US OFF FROM OUR PATH OF
ACHIEVING A CERTAIN GOAL, WE
MAKE AN ERROR.
FROM OUR PREVIOUS WORK, WE
IDENTIFY NEURONS IN THE HUMAN
MEDIAL FRONTAL CORTEX THAT CAN
DETECT AND SIGNAL THESE
DISTURBANCES ALMOST IMMEDIATELY
AFTER THEY OCCUR, BUT NOW THE
QUESTION IS, CAN THE SAME SET OF
NEURONS DETECT CONFLICT AND
ERROR ACROSS DIFFERENT TASKS OR
DO WE HAVE SPECIALIZED NEURONS
FOR EACH TASK.
THIS IS AN IMPORTANT QUESTION
BECAUSE UNLIKE ARTIFICIAL
INTELLIGENCE AGENTS, HUMANS HAVE
THIS AMAZING ABILITY TO
GENERALIZE.
WE CAN PERFORM MANY DIFFERENT
TASKS AND DEAL WITH ERRORS
DURING THE COURSE OF PER PERFE
IN ALL OF THESE TASK.
EVEN THOUGH THE SOURCES OF
DISTRACTION CAN DIFFER GREATLY.
MOTIVATED BY SUCH OBSERVATION,
WE TRY TO INVESTIGATE THE NEURAL
MECHANISMS THAT ALLOW THE MEDIAL
FRONTAL CORTEX TO SUPPORT
DOMAIN -- MONITORING.
WE RECORD SINGLE NEURONS FROM
EPILEPSY PATIENTS THAT WERE
MONITORED FOR SEIZURES TO SEE
VAL WAIT WHETHER
EVALUATE WHETHER THEY'RE A CAN'T
DATE FOR THERAPY.
IN TWO EXPERIMENTAL SESSIONS
WHILE RECORDING FROM THEIR
NEURONS.
WHAT WE FOUND IS THAT A LARGE
PERCENTAGE OF MEDIAL FORM OF
CORTICAL NEURONS INDEED ARE
DOMAIN GENERAL -- CONFLICTS AND
ERRORS IN BOTH TASKS.
SOME NEURONS ARE TASK SPECIFIC.
FOR EXAMPLE, THE SIGNAL ERRORS
WHEN THE PATIENT -- THE FIRST
TASK BUT -- SIGNALING ERROR TO
SWITCH TO A DIFFERENT TASK.
THIS HIGHLIGHTS THE PERFORMANCE
MONITORING CAN BE RECONFIGURED
RAPIDLY.
IMAGINE THE DOWNSTREAM
NEURONS -- WHETHER AN ERROR
OCCURS IN EITHER TASK REGARDLESS
OF THE REASONS FOR FAILURE, AND
ONLY LISTEN TO DOMAIN GENERAL
ERROR NEURONS BUT THIS IS
IMPOSSIBLE BECAUSE THEY ARE
INTERMIXED WITH A WHOLE LOT OF
OTHER NEURONS THAT ARE
TASK-SPECIFIC.
SO WHAT DOES DO THEN?
THE DOWNSTREAM NEURON CAN LEARN
THE SET OF WEIGHTS TO ASSIGN TO
THE ACTIVITY OF THE MEDIAL
FRONTAL CORTICAL NEURONS AND
COMPUTE A WEIGHTED SUM TO
EXTRACT INFORMATION, A PROCESS
WE CAN MIMIC USING LINEAR --
SUCH AS SUPPORT VECTOR MACHINES
IN MOTION LEARNING.
WE SHOW WE CAN BUILD LINEAR
CLASSIFIERS TO DECODE CONFLICTS
AND ERRORS FROM THESE NEURONS
WITH GREATER THAN 90% ACCURACY
ACROSS BOTH TASKS.
THIS RESULT SHOWS THAT THE
MEDIAL FRONTAL CORTEX IS ABLE TO
SUPPORT PERFORMANCE MONITORING
IN A DOMAIN-GENERAL WAY, AND
IT'S CRITICAL FOR COGNITIVE
FLEXIBILITY AS WE NAVIGATE
THROUGH DIFFERENT TASKS IN OUR
DAILY LIFE.
I HOPE YOU WILL BE CHECKING OUT
OUR POSTER, WHERE YOU CAN FIND
MORE INFORMATION AND DETAILS
ABOUT THIS WORK.
THANK YOU SO MUCH FOR LISTENING.
>> HI.
MY NAME IS LANDON BAYLESS
EDWARDS, I'M AN MD PH.D. STUDENT
AT OREGON HEALTH AND SCIENCES
UNIVERSITY.
OUR LAB STUDIES HOW
NEUROMODULATORS SUCH AS
DOPAMINE, SAIR TOE NEN AND NOR
NOREPINEPHRINE ALLOW US TO DO
THINGS LIKE LOAK MOTOR, SOMETIME
LIE SUCH AS TASTE OR SMELL.
THESE NEUROMODULATORS SIGNAL BY
BINDING TO RECEPTORS ON NEURONS
CALLED G PROTEIN COUPLED
RECEPTORS.
ALTHOUGH THERE ARE HUNDREDS TO A
THOUSAND DIFFERENT TYPES OF
THESE RECEPTORS, THEY ALL
TRANSMIT A SIGNAL BY MODULATING
THE CONCENTRATION OF THE SMALL
MOLECULE CYCLIC AMP.
THEREFORE WE WISH TO VISUALIZE
THESE DYNAMICS IN VIVO AS A
READOUT OF NEUROMODULATORY
SIGNALING.
WE CAN THEN DETERMINE THE
NEUROMODULATORS AT PLAY BY
COMBINING THIS APPROACH WITH
PHARMACOLOGY.
TO DATE, APPROXIMATELY 80 CYCLIC
AMP SENSORS HAVE BEEN DEVELOPED,
HOWEVER, NONE OF THESE SENSORS
HAVE BEEN USED AT A SINGLE
NEURON RESOLUTION IN VIVO DUE TO
LIMITED DYNAMIC RANGE AND LOW
SENSITIVITY.
TO OVERCOME THESE OBSTACLES, WE
FIRST COMPARED A SUBSET OF
EXISTENT CYCLIC AMP SENSORS IN
IDENTICAL CONDITIONS.
THE MOST PROMISING SENSOR FOR
USE IN VIVO WAS H187 AS IT
ALREADY HAD A LARGE DYNAMIC
RANGE.
-- TO IMPROVE UPON THE
SENSITIVITY AND SUBCELLULAR
DISTRIBUTION OF THIS SENSOR,
RESULTING IN A SERIES OF SENSORS
THAT WE CALLED CAMPFIRES, BASED
ON EPEC.
IN THE FIGURE ON THE BOTTOM
LEFT, I'M SHOWING A COMPARISON
OF A SUBSET OF PREVIOUSLY
PUBLISHED SENSORS COMPARED TO
THE CAMPFIRE SERIES.
WE APPLIED NOREPINEPHRINE AND
IBMX TO HEK293 CELLS THAT WERE
TRANSIENTLY EXPRESSING --
DYNAMIC RANGE RESPECTIVELY.
IN LIGHT BLUE HAVING THE HIGHEST
DYNAMIC RANGE.
YOU CAN SEE THAT THE CAMPFIRE
SENSORS SHOWN IN REDS AND ORANGE
HAVE INCREASED SENSITIVITY AND
ARE NAMED AS SUCH.
CAMPFIRE L LOWER SENSITIVITY, M
MEDIUM, AND H HIGH.
BUT THESE SENSORS ALL MAINTAIN
THE HIGH DYNAMIC RANGE OF THE
ORIGINAL EPAC SENSOR.
IN THE FIGURE ON THE BOTTOM
RIGHT, I'M SHOWING EXPRESSION OF
THE SENSOR IN NEURONS IN VIVO,
AND YOU CAN SEE THAT THE SENSOR
IS DISTRIBUTED DUE TO MUTATIONS
WE PUT IN THE NUCLEAR
LOCALIZATION MOTIF.
WE'VE SUCCESSFULLY USED THESE
SENSORS IN MULTIPLE PREPARATIONS
INCLUDING INCLUDE TOURED HIPPO
CAM PAP SLICES AND IN VIVO USING
A GRAIN WALL YAL CRANIA.
IMPORTANTLY, LIFETIME IMAGING IS
IDEAL FOR USE IN VIVO.
WE'RE ABLE TO MEASURE
DIFFERENCES IN CYCLIC AMP LEVELS
IN -- AND DENDRITES.
YOU CAN ALSO SEE THIS IN THE
FIGURE ON THE BOTTOM RIGHT.
THIS IS PARTICULARLY IMPORTANT
FOR IMAGING OF CYCLIC AMP, AS
IT'S DIFFERENTIALLY REGULATED IN
DIFFERENT SUBCELLULAR
COMPARTMENTS.
WE'VE ALSO SHOWN BI-DIRECTIONAL
CYCLIC AMP DYNAMICS IN VIVO.
MY EXAMPLE ON THE RIGHT SHOWS
THIS CHANGE AS WELL, WHERE THE
CELLS ON THE TOP SHOW AN
INCREASE IN CYCLIC AMP
CONCENTRATION DURING RUNNING ON
THE RIGHT COMPARED TO BASELINE
ON THE LEFT.
THE CELLS ON THE BOTTOM SHOW A
DECREASE IN CYCLIC AMP IN
RUNNING COMPARED TO BASELINE ON
THE LEFT.
FINALLY, WE SHOWED A REDUCTION
IN CYCLIC AMP IN RESPONSE TO
SYSTEMIC OPIOID ADMINISTRATION.
ALTHOUGH THIS HAS BEEN SHOWN EX
VIVO AND IN VITRO, IT HAD YET TO
BE SHOWN IN VIVO.
THIS FINDING IN PARTICULAR IS
EXCITING TO ME BECAUSE IT'S THE
FOUNDATION FOR MY FUTURE
DISSERTATION WORK.
WE'RE EXCITED TO SHARE THESE
SENSORS WITH ANYONE INTERESTED
IN IMAGING NEUROMODULATORY EVENT
AND ARE WORKING WITH CLAB RAY
FORS TO GENERATE A MOUSE LINE TO
MAKE THESE SENSORS MORE WIDELY
ACCESSIBLE.
THANK YOU.
>> HI, EVERYONE.
MY I'M A POSTDOC AT UC BERKE LE.
THE WORK I WILL BE PRESENTING
TODAY IS DEVELOPMENT OF
MULTIPHOTON HOLE GRAPHIC
OPTOGENETICS THAT ESSENTIAL RI
ALLOWS YOU TO DO OPTOGENETIC
PERTURBATIONS WITH SINGLE CELL
SPATIAL PRECISION.
SO IN OTHER WORDS, INSTEAD OF
STIMULATION OF THE WHOLE GENETIC
REDEFINED POPULATION OF NEURONS
LIKE YOU WOULD DO WITH
CONVENTIONAL OPTOGENETICS, HERE
WE'RE ABLE TO SIMULATE TARGETED
FUNCTIONALLY DEFINED ENSEMBLES
THAT CAN BE INTERMINGLED
SPATIALLY.
SO WHAT WE DID IN THIS PROJECT
IS TO PUSH THE LIMITS IN TERMS
OF SPEED AND FIDELITY OF
READ-WRITES.
SO WE USED THIS NEW CONSTRUCT
WHICH COMBINES BOTH SUPER POTEN
POTENT -- ME ETF WITH MP # M.
WITH THIS NEW CONSTRUCT, WE WERE
ABLE TO WRITE IN OPTOGENETIC --
READ THEM OUT WITH HIGH
RESOLUTION, SO THE CHALLENGE
NOW -- PARADIGMS IN 3D IS THAT
YOU CAN'T USE TYPICAL
SEQUENTIAL -- WITH THE --
ACTUATOR, FOR INSTANCE, BECAUSE
THE FRAME RATE IN THAT CASE
WOULD BE DIVIDED BY THE NUMBER
OF -- YOU TAKE.
SO TO ADDRESS THIS, WE ADDRESSEE
GRAPHIC HOLO
GRAPHIC MICROSCOPE AND ON THE
OTHER ARM HIGH SPEED VOL VOLUC
VESSEL BEAM IMAGING.
VESSEL BEAMS ARE THESE
ACTUALLY -- POINT SPREAD
FUNCTIONS, WHICH ALLOW YOU TO
RECORD THE PROJECTION OF THE
FULL VOLUME ON TO A SINGLE
CLAIM, ESSENTIALLY GIVING YOU
VOLUMETRIC RECORDING, BUT AT
SINGLE FRAME RATE.
SO WE TESTED AND VALIDATED OUR
MICROSCOPE AND SHOW WE ARE ABLE
TO WRITE IN AND READ OUT THESE
HIGHLY STRUCTURED TRAINS IN
POPULATIONS OF NEURONS IN FULL
3D.
JUST A QUICK COMMENT ON THESE
FREQUENCIES WE NOW HAVE ACCESS
TO 10 HERTZ, 20 HERTZ, UP TO 30
HERTZ IN SOME CASES.
THIS IS PARTICULARLY RELEVANT
SINCE IF YOU TAKE, FOR INSTANCE,
MOUSE -- NEURONS IN VISUAL CORE
TECH, THEY FIRE AN AVERAGE
AROUND 10 HERTZ.
SO NOW WITH THIS LARGE SCALE
DATABASE OF HIGHLY STRUCTURED --
WITH KNOWN HOLOGRAPHIC RUN
THROUGH WE CAN BENCHMARK BUT
ALSO INFORM NEW SPIKE INFERENCE
MODELS FOR A MORE PRECISE --
SPIKE RATES AND SPIKE TIMES.
AND SOME OTHER EXCITING
EXPERIMENTS WE'RE DOING RIGHT
NOW THAT WE'RE PRESENTING VISUAL
STIMULI TO AWAKE MICE AND
ATTEMPT TO FAITHFULLY CREATE
THOSE VISUALLY EVOKED SPIKE
TRAINS WITH VERY HIGH PRECISION.
SO THIS METHOD REALLY HOPES THE
DOOR TO ULTRA SIZE CUSTOM
TAILORED CORTICAL PROSTHETICS.
THAT'S IT FOR ME.
I'M HAPPY TO SHOW YOU MORE OF
THIS ON MY POSTER.
THANK YOU FOR YOUR ATTENTION.
>> HELLO, EVERYONE.
MY NAME IS MOHAMMAD, I'M A PH.D.
CANDIDATE AT U.C. SAN DIEGO AND
RESEARCH ASSISTANT.
MEMORIES ARE STORED ACROSS
NEURAL CIRCUITS -- PASSING THE
INFORMATION FLOW ACROSS NEURAL
CIRCUITS AND ALLOW
COMMUNICATIONS BETWEEN PRE AND
POST SYNAPTIC SITES BY MEANS OF
SYNAPTIC STRENGTH.
DENDRITIC -- VOLUMES ARE KNOWN
TO HAVE HIGH CORRELATION THE
POST SYNAPTIC DENSITY AREA,
ACTIVE ZONES AREA AND SYNAPTIC
STRENGTH.
SO WE USE THE SPINE HEAD VOLUMES
AS THE SURROGATE OF SYNAPTIC
STRENGTH IN THIS PROJECT.
THE MOTIVATION BEHIND THIS
PROJECT IS THE OBSERVATION OF
SUBSTANTIAL LIABILITY IN SNAPS
SIZE ES THAT RAISE THIS
QUESTION.
HOW DOES THE BRAIN RETAIN STABLE
MEMORIES ACROSS NEURAL CIRCUITS
AND HOW DO WE LEARN NEW THINGS
DESPITE HIGH INHERENT DYNAMICS
OF SYNAPTIC STRENGTH.
WE BELIEVE IT HIGHLY DEPENDS ON
THE PRECISION OF SYNAPTIC
PLASTICITY.
TO FURTHER EXPLORE THIS
HYPOTHESIS, WE HAVE USED THREE
DIMENSIONAL RECONSTRUCTION OF
SERIAL SECTION ELECTROMICROSCOPY
THAT PROVIDES HIGH RESOLUTION
AND DETAILS OF PRE AND POST
SYNAPTIC MORPHOLOGY AND --
RESOURCES THAT IS NOT FEASIBLE
IN OTHER TECHNIQUES SUCH AS
LIGHT MICROSCOPY.
HERE IN THE TOP GRAPH, YOU SEE
THE LARGEST AND THE SMALLEST
SPINE HEAD VOLUMES MAINTAIN EACH
OF THE DATASETS AND THE RANGE OF
THEIR VOLUME IN THE ROLE
DENOTED.
THIS CLEARLY SHOWED THAT THE
INDUCTION OF LONG TERM
POTENTIATION EXPANDED THE RANGE
IN COMPARISON TO THE CONTROL --
FINALLY, IN THE BOTTOM FOUR
SUBPLOTS, YOU SEE THE NUMBER OF
DISTINGUISHABLEAPTIC
STRENGTH -- EACH OF THE FOUR
DATASETS.
AS YOU CAN SEE BY COMPARING
PANEL A TO C, 30 MINUTES AFTER
INDUCTION OF LTP, THE NUMBER OF
SYNAPTIC STRENGTH IS DOUBLED AND
SHOWN BY THE TWO HOURS POST
INDUCTION OF LTP, THEY STAYED
HIGHER THAN THE CONTROL CASES.
FURTHERMORE, OUR DIVERGENCE
ANALYSIS REVEALS THAT AT THE
TWO-HOUR POST INDUCTION OF LTP,
THE DISTRIBUTION OF CLUSTERS IS
MOVED CLOSER TO THE UNIFORM
DISTRIBUTION, IMPLYING EFFICIENT
CODING OCCURS AFTER LEARNING.
THANK YOU SO MUCH FOR YOUR TIME,
AND PLEASE COME TO MY POSTER FOR
FURTHER DISCUSSION.
>> HI.
MY NAME IS LUISA SHUSTER,
GRADUATE STUDENT AT NEW YORK
UNIVERSITY AND TODAY I WILL BE
DISCUSSING MY PROJECT OPEN
SOURCE HIGH-THROUGHPUT
MONITORING SYSTEM TO MATERNAL
BEHAVIOR MICE.
-- CRUCIAL FOR THE SURVIVAL OF
SPECIES.
TYPICALLY THE FIRST SOCIAL
INTERACTION IN ANIMAL
EXPERIENCES.
-- MAMMALS AND BECAUSE OF ITS
RICHNESS AND COMPLEXITY IN ORDER
TO UNDERSTAND ITS NEURAL
UNDERPINNINGS, NECESSARY TO
STUDY THESE INTERACTIONS
COMPREHENSIVELY.
TO ACHIEVE THIS, I DEVELOPED A
LOW COST OPEN SOURCE ENCLOSURE
TO LOOK AT THE NATURAL
PROJECTION OF MOTHERHOOD UP
UNTIL THREE TO FOUR CONSECUTIVE
ROUNDS OF BREEDING.
USING DIFFERENT SENSORS SUCH AS
CAMERAS, THERMAL SENSORS, IT
GIVES US INSIGHTS INTO THE
SECRET LIVES OF MICE AND HOW
THEY'RE INTERACTING WITH THEIR
THOUGHTS.
HOWEVER, THIS HAS BEEN USED FOR
THE TYPES OF SOCIAL INTERACTIONS
SUCH AS MALE/FEMALE --, ET
CETERA.
SOME OF THE THINGS WE'VE BEEN
ABLE TO FIND USING THIS APPROACH
THAT WE WOULD NOT HAVE BEEN ABLE
TO FIND HAD WE BEEN USING --
SUCH AS -- JUST A SNAPSHOT OF
PARENTAL BEHAVIORS IS THAT WHEN
WE ANALYZE THE -- OF ANIMALS
DURING PREGNANCY, WE NOTICE THAT
DEPENDING ON -- INTEGRITY SCORE
IF IT WAS LOW, -- MEANING THAT A
LOT OF PUPS COULD DIE.
HOWEVER, IF THE QUALITY SCORE
WAS HIGH, THE ANIMALS WERE
LIKELY TO SURVIVE POST -- UP TO
100%.
WE DECIDED TO LOOK TETHER MALL
PROPERTIES OF THIS NEST, HOW
THEY WERE CONTRIBUTING TO THE
WARMTH OF THE PUPS, AND WE SAW
THAT FEE SURES SUCH AS THE A
WERE SUPER IMPORTANT IN
DETERMINING HOW -- A NEST IS,
EVEN THOUGH THE PUPS CANNOT
THERMOREGULATE AND THEY REQUIRE
CONSTANT INSULATION PROVIDED BY
THE MOM OR THE NEST AND THE MOM
MATES.
WITH MY PROJECT, I AM HOPING TO
LOWER THE -- BARRIERS -- EVEN
SOCIAL BEHAVIORS ARE VERY
EXPENSIVE TO STUDY.
>> HELLO, EVERYONE.
I AM A RESEARCH ASSOCIATE AT THE
BIOMEDICAL ENGINEERING
DEPARTMENT OF UNIVERSITY OF
SOUTHERN CALIFORNIA.
THANK THE NIH BRAIN INITIATIVE
FOR GIVING ME THIS GREAT
OPPORTUNITY TO SHARE SOME OF MY
WORKS WITH YOU.
COMMON MICRO ELECTRODE ARRAYS
ARE FLEXIBLE AND CAN GREATLY
REDUCE MECHANICAL MISMATCH
BETWEEN BRAIN TISSUE AND NEURAL
IMPLANT.
THIS MAKES IT A VERY PROMISING
TOOL FOR CHRONIC MONITOR AND
INTERFACING WITH NEURON TO
BETTER PROMOTE THE DEVELOPMENT
AND -- OF POLYMER MEAs.
WITH NIH'S SUPPORT, PROFESSOR
ALICE LONG AND PROFESSOR DONG
SONG LAUNCHED THE -- TO PROVIDE
NEUROSCIENCE AND
NEUROENGINEERING COMMUNITY WITH
AN EASY ACCESS TO FLEXIBLE
POLYMER MEAs TO GIVE
RESEARCHERS A CHOICE OF READY TO
USE POLYMER MEAs BY DESIGNING
A PENETRATING ARRAY -- WHICH CAN
BE USED TO RECORD FROM CORTICAL
AND SUBCORTICAL REGIONS OF
RODENT BRAINS.
THIS MEA HAS 64 ELECTRODES WITH
15 -- DIAMETERS TO OBTAIN NEURAL
RECORDINGS WITH SINGLE CELL
RESOLUTION.
CONSIDERING THE ANATOMY OF RAT
AND MICE BRAINS, THE MEA IS
DESIGNED TO HAVE -- SO THAT IT
CAN REACH SUBCORTICAL STRUCTURES
SUCH AS HIPPOCAMPUS.
WITH 350 MICRO METER SPACING
BETWEEN SHANKS, A RELATIVELY
LARGE BRAIN AREA OR MULTIPLE
CORTICAL REGIONS CAN BE COVERED
ON EACH SHANK, 16 ELECTRODES
WITH 120 MICRO METER SPACING BY
EVENLY DISTRIBUTED AT THE TIP
WITH 1.8-MILLIMETER RECORDING
RANGE, THE MEA CAN RECORD FROM
MULTIPLE LAYERS OF THE CORTEX OR
MULTIPLE LAYERS OF THE
HIPPOCAMPUS SIMULTANEOUSLY.
TO ASSIST THE IMPLANTATION OF
FLEXIBLE MEAs, I ALSO
DEVELOPED A VERY PRACTICAL
DEEP -- PRACTICAL PROTOCOL FOR
INSERTION OF THIS GENERIC
NEUROPROBE.
FOLLOWING THIS PROTOCOL, THE MEA
CAN BE TEMPORALLY -- BY A SINGLE
LAYER OF POLYETHYLENE -- WITH NO
SPECIFICALLY FABRICATED --
INVOLVED.
THIS PROTOCOL CAN GREATLY EASE
THE IMPLANTATION OF FLEXIBLE
MEAs.
TO TEST THE DURABILITY OF THE
COATING METHOD AND TO
CHARACTERIZE THE PERFORMANCE OF
THE POLYMER MEA, SIX PROBES
COATED WERE IMPLANTED INTO THE
RAT HIPPOCAMPUS.
ALL MEAs WERE SUCCESSFULLY
INSERTED INTO THE HIPPOCAMPUS,
AND RECORDED LOCAL FIELD
POTENTIALS SUCH AS -- WAVES AND
ACTIVITIES WITH SIGNAL TO NOISE
RATIO GREATER THAN 3.
FOR TWO ANIMALS, OVER 30 UNITS
WERE RECORDED FROM MULTIPLE
LAYERS OF THE RAT HIPPOCAMPUS ON
THE IMPLANTATION DAY.
PRELIMINARY EVALUATION OF THE
CHRONIC PERFORMANCE OF THE
POLYMER MEA WAS ALSO CONDUCTED.
HIGH AMPLITUDES, THETA WAVES AND
SPIKES WERE OBSERVED OVER A
PERIOD OF 37 DAYS.
AND THE SAME -- RECORDED UNITS
WAS SIMILAR WAVEFORMS OVER A
PERIOD OF 34 DAYS.
THESE PRELIMINARY EVALUATIONS
INDICATE THAT THIS GENERIC MEA
CAN SERVE A VALUABLE TOOL FOR
THE STUDY OF BRAIN FUNCTION.
THANKS A LOT FOR YOUR TIME AND
I'M MORE THAN HAPPY TO DISCUSS
ANY DETAIL DURING THE Q & A
SESSION.
>> HELLO.
MY NAME IS BEN, PH.D. CANDIDATE
AT JOHNS HOPKINS UNIVERSITY
WORKING IN JOSHUA VOGELSTEIN'S
LAB.
I WORK ON STATISTICAL AND
COMPUTATIONAL METHODS FOR MAPS
OF CONNECTIVITY.
HOW THE BRAIN CHANGES ACROSS
DEVELOPMENT, EVOLUTION,
EXPERIENCE, OR DISEASE STATE.
TO UNDERSTAND THESE PROPERTIES
AND HOW THEY RELATE TO NEURAL
WIRING, RESEARCHERS ARE
BEGINNING TO MAP MULTIPLE
ASSOCIATED CONNECTOMES, SAY FROM
TWO DIFFERENT RELATED SPECIES,
AND THEN COMPARE THESE
CONNECTOMES TO UNDERSTAND WHICH
ASPECTS OF NEURAL WIRING ARE
PRESERVED OR NOT.
MY WORK FOCUSES ON HOW TO MAKE
THESE ASSESSMENTS ONCE WE HAVE
THE DATA.
TWO MAPS OF NEURAL WIRING WILL
ALMOST NEVER BE IDENTICAL.
SO HOW CAN WE SAY WHETHER ANY
DIFFERENCES WE OBSERVE BETWEEN
THEM ARE MEANINGFUL?
AND WHAT DO WE EVEN MEAN BY
DIFFERENT FOR DIFFERENT
INTERCONNECTED NEURONS.
WE DEVELOPED A NEW METHODOLOGY
TO ANSWER THESE QUESTIONS BASED
ON BOTH WORK FROM NETWORK
SCIENCE WHICH STUDIES
CONNECTIVITY AND STATISTICS
WHICH STUDIES HOW TO MAKE CLAIMS
ABOUT WHETHER DIFFERENCES
BETWEEN TWO THINGS ARE MEANING
FU.
MEANINGFUL.
AN EXAMPLE OF THIS KIND OF
COMPARISON IS SHOWN TO THE
RIGHT.
HERE WE PLOT THE CONNECTIVITY OF
THE LECHT AND LEFT AND RIGS
OF BRAIN OF A DROSOPHILA OR
VINEGAR FLY DURING ITS LARVAL
STAGE.
MORE IMPORTANTLY, WE DEVELOPED
STATISTICAL THEY MODS FOR METHO.
IN THE LARVA, WE FOUND SPECIFIC
NEURON TYPES DISPLAY DIFFERENT
CONNECTIVITY PATTERNS TWEAN THE
TWO BRAIN HEMISPHERE, BUT ALSO
THAT THIS DIFFERENCE GOES AWAY
IF ONE ONLY CONSIDERS THE
STRONGEST NEURAL CONNECTIONS.
THESE FINDINGS SUGGEST WHICH
PROPERTIES OF THE NEURAL WIRING
ARE IMPORTANT FOR THE GENOME TO
PRESERVE ON BOTH SIDES OF THE
BRAIN OF THIS ORGANISM.
AND WHILE OUR MORE RECENT WORK
APPLIED THESE TOOLS TO STUDY THE
BILATERAL SYMMETRY OF THE SINGLE
CONNECTOME, I'M IK SEATED O
COMPARE OTHER CONNECTOMES IN A
PRINCIPAL WAY.
WHILE WE'RE STILL CONTINUING TO
EXPAND ON DIFFERENT NOTIONS
BETWEEN NETWORKS WHICH WE CAN
DETECT, WE EXPECT THIS FRAMEWORK
FOR STATISTICAL COMPARISON OF
CONNECTOMES WILL BE A AN
IMPORTANT COMPONENT TO INFORM
OUR IS STUDY OF THE BRAIN.
SPECIFICALLY HOW NEURAL
CONNECTIVITY IS RELATED TO THE
MANY PROPERTIES I MENTIONED LIKE
DEVELOPMENT EVOLUTION,
EXPERIENCE, OR DISEASE STATE.
TO THAT END, WE'VE DEVELOPED
OPEN SOURCE PYTHON CODE TO
PERFORM ALL OF THESE ANALYSES
ALONG WITH ONLINE NOTEBOOKS TO
GUIDE RESEARCHERS THROUGH THE
PROCESS OF USING THIS CODE.
WE'RE ALWAYS HAPPY TO HAVE NEW
COLLABORATORS, SO PLEASE REACH
OUT IF ANY OF THIS IS
INTERESTING OR IF YOU HAVE
CONNECTOMES THAT YOU WANT TO
COMPARE.
THANK YOU.
>> WONDERFUL.
THANK YOU ALL, CONGRATULATIONS
AGAIN TO ALL OF OUR TRAVEL
AWARDEES IN THIS SESSION, AS
WELL AS THE OTHER CONCURRENT
SESSION THAT IS GOING ON.
THOSE WERE GREAT TALKS.
WE DO HAVE SOME QUESTIONS THAT
HAVE BEEN SUBMITTED THROUGH THE
Q & A FUNCTION AND WE'LL JUST
REMIND OUR AUDIENCE THAT THERE
IS STILL TIME TO SUBMIT
QUESTIONS FOR ANY OF OUR
SPEAKERS TODAY.
WE WILL GO THROUGH THEM.
WE HAVE ABOUT 30 MINUTES OR SO
TO GET THROUGH QUESTIONS.
SO I'M JUST GOING TO GO AHEAD
AND START AND JAY AND I WILL
SORT OF TAG TEAM PRESENTING THE
QUESTIONS TO OUR SPEAKERS.
THE FIRST ONE WILL GO TO
DESIREE.
CAN DANCO GIVEN PERIPHERALLY AND
THEN UNCAGED LOCALLY IN THE
BRAIN OR MUST IT BE GIVEN
LOCALLY?
>> HI.
THANK YOU SO MUCH FOR YOUR
QUESTION.
DANCO IS A PEPTIDE.
BECAUSE OF THAT, IT'S SUBJECT TO
CLEARANCE BY PEPTIDASES SO IT
NEEDS TO BE GIVEN LOCALLY IN THE
BRAIN.
OTHER SMALL MOLECULE -- CAGED
SMALL MOLECULES SUCH AS THE
OXIES IS MORPHONE CAN BE GIVEN
SYSTEMICALLY AND UNCAGED LOCALLY
IN THE BRAIN REGION OF INTEREST,
AND IF YOU'RE INTERESTED IN
THAT, MY POSTER IS NUMBER 320
AND CAN YOU CHECK IT OUT.
BUT DANCO NEEDS TO BE
ADMINISTERED LOCALLY, YES.
>> THANKS FOR THAT, DESIREE.
I'LL JUST MAKE A QUICK COMMENT.
THE BEST WAY THAT I CAN DESCRIBE
WHAT WE JUST HEARD IS, WOW!
I MEAN, THE DIVERSITY OF
SCIENCE, THE PERSPECTIVES, THE
EXPERIENCES FROM THE PANELISTS
THAT WE HEARD ABOUT, IT JUST
SUGGESTS THAT THE FUTURE OF
BIOMEDICAL RESEARCH IS LOOKING
HUGELY PROMISING.
SO AGAIN, CONGRATULATIONS TO ALL
OF YOU.
AS A REMINDER, FOR EVERYBODY
BEYOND OUR PANELISTS, ALL OF THE
RESEARCH HIGHLIGHTS YOU JUST
HEARD FROM ARE BEING PRESENTED
AS POSTERS AS WELL.
SOME OF THEM MAY HAVE BEEN
PRESENTED ALREADY, THERE ARE
MORE COMING, SO I ENCOURAGE YOU
TO VISIT THESE POSTERS AND HAVE
ADDITIONAL CONVERSATIONS WITH
PRESENTERS.
I'LL SAY THAT I'VE GOT GENERALLY
TWO BUCKETS OF QUESTIONS.
ONE ARE SOME VERY SPECIFIC
RESEARCH QUESTIONS THAT PEOPLE
HAVE SUBMITTED AND WE WILL COVER
AS MANY OF THESE AS WE CAN, AND
THERE'S ANOTHER GENERAL BUCKET
OF SORT OF JUST LIFE
EXPERIENCES, PARTICULARLY AT
YOUR CAREER STAGE.
SO WE OPEN THIS UP TO ALL
PANELISTS IF YOU HAVE SOMETHING
TO SAY.
BUT WITH THAT, I'LL JUMP INTO
ONE OF THE QUESTIONS THAT WE DID
RECEIVE.
THIS ONE IS FOR HUNYUN SHIN,
HAVE YOU DONE ANY BEHAVIORAL TWO
CHOICE TASK BETWEEN THE ILLUSORY
CONTOUR AND THE PROBE CONTOURS
TO SEE IF THE ILLUSION
PERCEPTION HOLDS BEHAVIOR --
HOLDS IN BEHAVIOR?
>> YES, THANK YOU FOR THE
QUESTION AND SORRY I DIDN'T
COVER THE PRIOR LITERATURE, BUT
THERE HAS BEEN TWO PRIOR STUDIES
USING MICE, USING ALTERNATIVE
POOR CHOICE BEHAVIOR SHOWING
MILES CAN -- I LEWIS ILLUSOD
VICE VERSA SO THEY CAN BE
PERCEIVED BY MICE.
SO BUILDING UPON THAT, OUR STUDY
FOCUSED ON THE REPRESENTATION
INDIVIDUAL PROTECTION ESPECIALLY
FOR HIGHER VISUAL AREAS, THIS IS
THE FIRST STUDY TO LOOK AT
HIGHER VISUAL AREA
REPRESENTATION IN MICE.
>> I HAVE A QUICK FOLLOW-UP
QUESTION FOR YOU AS WELL.
YOU MAY HAVE SAID THIS BUT I
MISSED IT.
WHAT AGES ARE YOU TESTING AND IS
THERE SOME PREDICTABLE
FUNCTIONAL PATTERNS IN TERMS OF
DEVELOPMENT WHEN THESE SYSTEMS
COME ONLINE?
>> THAT'S A GREAT QUESTION, ONE
THAT I CANNOT ADDRESS.
SO I'M JUST USING ADULT MICE AND
I HAVEN'T LOOKED AT --
DEVELOPMENT.
BUT YEAH, IT'S LEARNING AND
EXPERIENCES IS IMPORTANT FOR
PERCEIVING THE SOLUTION, YOU
WOULD THINK THAT WOULD CHANGE --
DEVELOPMENT SO THAT WOULD BE A
FUTURE AVENUE.
>> SOUNDS GOOD.
THIS ONE IS FOR MICHELLE.
WHAT ARE YOU USING TO DELIVER
THE JEDI-2P INTO THE BRAIN?
IS IT AAVs, CAN THEY BE
TARGETED TO SPECIFIC CELL TYPES?
>> GREAT QUESTION.
YEAH, THAT'S ONE OF THE BEAUTIES
OF GENETICALLY ENCODABLE
INDICATORS, THEY'RE ENTIRELY
GENETICALLY ENCODED SO YES, WE
USE AAVs TO GET THEM WHERE
THEY SHOULD BE.
WE HAVE MANY DIFFERENT
PROMOTERS, SO ONE OF THE ONES
THAT YOU PROBABLY SAW, SHOUT OUT
FOR THAT BEAUTIFUL DATA, IS A
KREE-DEPENDENT ONE THAT WE HAD
FOR EF1 ALPHA, AND BASICALLY --
SO WE HAVE CRE-DEPENDENT PLA
MASS, WE HAVE PROMOTERS THAT CAN
BE JUST NEURONAL SPECIFIC LIKE
HCIN, OR WE CAN DO SOMETHING
LIKE IM. GLUTAMATERGIC OR CELL
EXPRESSION.
NON-RODENT MODELS AS WELL LIKE
FLEAS.
IF YOU'RE INTERESTED, GO TO MY
POSTER, 814, AND WE HAVE SOME
WAYS TO CONTACT US THERE.
>> THANKS FOR THAT, MICHELLE.
I THINK ANYBODY WOULD WANT TO GO
HEAR MORE ABOUT JEDIs, OR THE
LAST OR MAYBE NOT THE LAST AS
WELL.
SO VERY CREATIVE.
NICE JOB.
SO WE HAVE ANOTHER QUESTION THAT
CAME IN.
THIS ONE IS FOR LANDON.
THE QUESTION IS, IS CAMPFIRE
COMPATIBLE FOR COIMAGING WITH
CALCIUM IMAGING?
>> YEAH, THAT'S A GREAT
QUESTION.
THERE IS POTENTIAL COMPATIBILITY
THERE.
THIS IS SOMETHING WE HAVEN'T
DONE YET, BUT IT'S SOMETHING
WE'RE PLANNING TO DO.
ONE OF THE BENEFITS OF CAMPFIRES
IS THAT THEY'RE COMPATIBLE WITH
FLUORESCENCE LIFETIME IMAGING,
SO THIS FREES UP A CHANNEL THAT
COULD BE USED, FOR EXAMPLE, WITH
A RED CALCIUM SENSOR.
>> SOUNDS GOOD.
I HAVE A QUESTION FOR NATALIE.
I JUST COULDN'T SEE THE SLIDE
VERY WELL SO THIS IS FOR MY OWN
CLARIFICATION.
WHAT WAS THE SCALE ON THE OXYGEN
CONSUMPTION WHERE YOU FIRST
NOTED THAT LAYER ONE WAS THE
HIGHEST IN TERMS OF OXYGEN
CONSUMPTION COMPARED TO LAYER
FOUR, WHICH WAS WHERE WE MIGHT
PREDICT TO BE SO, WHAT WAS THE
SCALE BETWEEN THE TWO?
>> IT WAS LIKE DOUBLE.
SO IN LAYER ONE, WE HAD DOUBLE
MORE OXYGEN CONSUMPTION.
>> AND CAN YOU -- WHAT'S YOUR
BEST GUESS AS TO WHY WE'RE
SEEING THIS IN LAYER ONE?
>> NOT MANY CELL BODIES, THERE
ARE A LOT OF DENDRITES AND WE
THINK DENDRITIC COMPUTATIONS
COST A LOT OF ENERGY.
SO THERE ARE LONG-RANGE
CONNECTIONS BETWEEN CORTICAL
AREAS, BUTS ARE ASCENDING
CONNECTIONS FROM BASAL
FOREBRAIN, CHOLINERGIC FIBERS
COME THERE FROM THE -- SO THERE
ARE MANY INTERCONNECTIONS GOING
ON, AND ALL -- THEY ARE 45, BUT
FOUR-FIVE BUT THEY PROJECT TO
LAYER ONE AND HAVE THIS
BEAUTIFUL TREE, SO WE THINK THIS
COSTS A LOT OF ENERGY.
BUT IN ADDITION, ONE THING TO
KEEP IN MIND IS THAT WHAT WE SEE
IS TRUE, MIGHT BE TRUE, HOWEVER,
UNDER CERTAIN CONDITIONS, LET'S
SAY UNDER DRASTIC CONDITIONS
WHEN OXYGEN LEVELS SINK -- OR
MAYBE DURING AGING WHEN OXYGEN
DELIVERY ISN'T AS EFFICIENT AS
IN YOUNG STATES, IT MIGHT BE
IMPORTANT TO HAVE HIGHER
MITOCHONDRIAL DENSITY AND
THEREFORE TO EXTRACT OXYGEN
BETTER.
THIS IS ONE IDEA.
BUT THIS IS SOMETHING THAT CAN
BE STUDIED FURTHER.
BUT IT'S DEFINITELY NOT JUST
UNDER RESTING CONDITION.
>> VERY GOOD.
THANK YOU.
WE ALSO HAVE A QUESTION FOR
CARLOS.
IF YOU DISRUPT THE AUDITORY
PROJECTING NEURONS DURING THE
LEARNING PHASE, DO THEY NOT
LEARN THE SONGS?
>> THANK YOU FOR THAT QUESTION.
SO RECENT WORK HAS SHOWN IF YOU
USE CRE-DEPENDENT -- ONLY THE
AUDITORY PROJECTING NEURONS
DURING THE LEARNING PHASE
SPECIFICALLY THEY WILL NOT LEARN
THEIR SONG.
THE DEGREE OF LEARNING
CORRELATES WITH THE AMOUNT OF
LESIONS, SO MORE CELLS --
PROJECTED TO THE AUDITORY
REGION, A DECREASED AMOUNT OF
LEARNING.
BUT IF YOU DO THIS IN THE ADULT
BIRDS WHEN THEY HAVE ALREADY
LEARNED THEIR SONG, IT WILL HAVE
NO AFFECT ON THEIR SONG.
SO THIS WORK ALSO HIGHLIGHTS THE
IMPORTANCE OF THESE SPECIFIC
CELL TYPES AT A PARTICULAR TIME
POINT IN DEVELOPMENT.
>> OKAY, THANK YOU, CARLOS.
LET'S SEE.
I'M GOING TO JUMP INTO ONE OF MY
OWN QUESTIONS I ACTUALLY HAVE
HERE, AND I WANT TO OPEN THIS UP
TO THE PANELISTS.
WE'RE GOING TO COME BACK TO SOME
MORE SPECIFIC SCIENCE QUESTIONS.
WITH YOU I'M BUT I'M REALLN
HEARING FROM THE PANELISTS AND
HAVING YOU SHARE YOUR
EXPERIENCES.
WE'VE ALL JUST GONE THROUGH
WHERE WE'RE GOING THROUGH THIS
THING WE CALL COVID, WHICH WE
COULD HAVE NEVER ANTICIPATED,
AND IT HAS HAD AN ABSOLUTE
IMPACT ON OUR LIVES, ON OUR
ABILITY TO DO SCIENCE,
PARTICULARLY FOR PEOPLE ON THE
PANEL HERE WHO ARE AT VERY
CRITICAL STAGES IN THEIR CAREER
DEVELOPMENT.
SO I'M CURIOUS IF YOU CAN OFFER
SOME OF WHAT YOU HAVE DONE FROM
A STRATEGIC PERSPECTIVE TO
REMAIN PRODUCTIVE, TO CONTINUE
TO MOVE FORWARD AND NOT JUST TO
STOP WHERE YOU'RE AT.
I THINK OTHERS WOULD BE REALLY
INTERESTED TO HEAR WHAT SOME OF
THOSE THINGS HAVE BEEN.
AND THAT'S OPEN TO ANYBODY.
>> I CAN START.
SO AS YOU KNOW, I DO
24/7 RECORDING, SO BEHAVIOR
ACROSS CONSECUTIVE -- SO WHEN
THE PANDEMIC STARTED AND WE -- I
HAD TO STOP EXPERIMENTS SO THAT
ACTUALLY INSPIRED ME TO DESIGN A
SYSTEM THAT COULD BE ACCESSED
REMOTELY AND IT WAS PRETTY MUCH
SELF-SUFFICIENT, OTHER THAN THE
HUSBANDRY WITH THE ANIMALS.
SO THE PANDEMIC PRETTY MUCH
INSPIRED WHAT MY PROJECT IS NOW,
AND TO CREATE AN ENCLOSURE WHERE
WE COULD HAVE THE ANIMALS FOR
LONG PERIODS OF TIME, THAT WAS
INEXPENSIVE TO MATE THAT COULD
BE ACCESSED REMOTELY BECAUSE --
YOU CAN BE IN YOUR PJs AT HOME
AND LOOKING THROUGH THE CAMERAS
AND SEE IF YOUR ANIMALS ARE
DOING WELL AND THE ANALYSIS CAN
BE RUN REMOTELY.
>> I CAN ADD TO THAT.
I DO IMPLANTS WHICH IS ITSELF
VERY HARD TO GET, AND THEN WITH
COVID, I THINK WE HAD LIKE 1 1/2
YEARS WITH NO -- BECAUSE IN THE
HOSPITAL IT'S SUPER CHAOTIC SO
NO ONE WANTS TO DO THAT.
SO I THINK ONE THING THAT HELPED
ME TO MOVE FORWARD WAS TO DIG
MORE INTO THE LITERATURE.
MAINLY WHEN SOMEONE IS WORKING
IN DISCIPLINARY SCIENCES, I
THINK IT'S NEVER ENOUGH TO
LIKE -- NEVER ENOUGH NO MATTER
HOW MUCH YOU KNOW THE -- YOU'RE
WORKING ON.
SO I THINK I TAKE A LOT INTO
THAT AND EVEN STARTED WRITING
PAPERS BEFORE WE HAVE DATA JUST
LIKE THE INTRODUCTION AND STUFF
SO YOU CAN GET A HEAD START AND
NOW WHEN THINGS ARE BACK
RUNNING, I THINK THEY ARE GOING
PRETTY FAST.
SO THAT HELPED ME AT LEAST
PERSONALLY A LOT.
>> OKAY.
WE CAN COME BACK TO THAT AT ANY
POINT IF SOMEBODY HAS ANYTHING
REALLY INSIGHTFUL THEY WANT TO
SHARE.
LET'S SEE.
LUISA, YOU SPOKE A MOMENT AGO
BUT I'M GOING TO PUT YOU BACK IN
THE SPOTLIGHT AGAIN HERE.
THERE WAS A QUESTION THAT CAME
IN FOR YOU, AND IT SAID, HAVE
YOU LOOKED AT ANY OF YOUR
COLLECTED DATA USING OPEN SOURCE
TOOLS, TOOLS LIKE DEEP LAB CUT
OR TO USE OTHER SYSTEMS FOR YOUR
VIDEO ANALYSIS PIPELINE?
>> THAT'S A GOOD QUESTION.
SO THE IDEA OF MY PROJECT IS TO
MAKE IT AS LOW COST AS POSSIBLE.
SO I'VE BEEN USING -- TO TRACK
THE -- OF THE MOM WHEN SHE'S OUT
OF THE NEST BECAUSE THAT WAY, WE
CAN SYNCHRONIZE IT WITH LIKE OUR
THERMAL CAMERA AND LOOK AT THE
TEMPERATURE OF THE PUPS.
WE HAVE ALSO ANOTHER NETWORK
TRAINED TO IDENTIFY WHEN THE
PUPS ARE OUT OF THE NEST, AND WE
HAVE ANOTHER ONE TO IDENTIFY
ANIMALS OF THE SAME SIZE.
AND ALSO USING THAT -- DATA
WE'RE GETTING FROM DEEP LAB CUT,
MORE RECENTLY I'VE BEEN USING
OPEN SOURCE BEHAVIORAL
CLUSTERING SOFTWARE DEVELOPED
BY -- BECAUSE EVENTUALLY WE WANT
TO AUTOMATE AS MANY BEHAVIORS AS
POSSIBLE BECAUSE OUR VIDEOS ARE
SO LONG THAT DOING IT MANUALLY
IS IMPOSSIBLE.
SO EVERYTHING IN OUR ANALYSIS
PIPELINE IS OPEN SOURCE AND IT'S
BASED ON OTHER PEOPLE'S WORK.
SO I APPRECIATE THAT QUESTION.
THANK YOU.
>> THANK YOU, LUISA.
SO OUR NEXT QUESTION IS FOR WUI
JENG.
THE QUESTION IS, DO THESE MEAs
RESULT IN LESS GLIA ACTIVATION,
SCARRING --/SCARRING, THAN MORE
TRADITIONAL ELECTRODES?
>> FIRST THANKS FOR THE
QUESTION.
SO THIS PARTICULAR ARRAY, WE
HAVEN'T DONE ANY LINEAR RESPONSE
STAINING YET, SO THE ONLY
HISTOLOGY WORK I DID ON THIS
PROBE IS A NASAL STAIN SO I DO
SEE VERY LAST SCAR OR LIKE
WHATEVER -- AROUND THOSE PROBES.
BUT WE DO HAVE EXPERIENCE WITH
ANOTHER MICRO ELECTRODES ARRAY
WITH THE SAME MATERIAL WITH VERY
SIMILAR -- FROM OUR PREVIOUS
WORK.
FROM THAT ONE, WE DO SEE LESS
ACTIVATION OF SCAR TISSUE AND
SCARRING AROUND THE PROBE.
IT'S NOT A, LIKE, DRAMATIC --
DRAMATIC-REDUCING BUT SLIGHTLY
LESS AND COMPARABLE TO LIKE
MICRO WIRES AND MUCH BETTER
THAN -- PROBES.
>> THANKS FOR THAT.
BENJAMIN HAS A COUPLE OF
QUESTIONS HERE.
THERE'S A BIT OF PA A PRIMACY
EFFECT HERE OR REESE SEE -E
INTENSIVE IS RUNNING THIS
CONNECTOME CONNECTION
COMPARISON, AND CAN IT BE RUN ON
A DESKTOP OR IS GP PROCESSING
NEEDED?
>> SO THANKS FOR THE QUESTION.
I GUESS THE SHORT ANSWER IS
EVERYTHING I'VE BEEN DOING, I
JUST RUN ON MY LAPTOP, SO IT'S
NOTHING WE'VE DONE SO FAR IS
VERY COMPUTATIONALLY EXPENSIVE.
IT TAKES -- MANY OF THESE TESTS
TAKE LESS THAN A SECOND.
SOME OF THE STUFF WE'RE WORKING
ON NOW, I THINK MIGHT END UP
BEING A BIT MORE COMPUTATIONALLY
EXPENSIVE, AND ALSO AS DATASET
GET LARGER, THAT MIGHT BE A
LITTLE BIT MORE OF A CONCERN,
BUT I DON'T THINK IT WOULD BE
PROHIBITIVE FOR RUNNING ANY OF
THESE COMPARISONS.
>> THANKS FOR THAT.
I WANTED TO ACTUALLY LOOK TO THE
PANELISTS AS WELL.
I SEE -- YOU'VE GOT YOUR HAND
UP.
IS THERE SOMETHING YOU WANT TO
JUMP IN ON HERE?
>> YEAH, I CAN HAVE A QUESTION
FOR BENJAMIN AS WELL.
IT A MORE BASIC ONE.
I THINK YOU MENTIONED IN YOUR
PRESENTATION THAT ONE OF THE
PURPOSE TO LOOK AT THIS IS TO
STUDY BETWEEN SPECIES, HOW THE
CONNECTOMES ARE SIMILAR.
I'M JUST CURIOUS WHAT DO YOU
THINK CAN BE THE OOS FULL USEFUS
OF THAT, AND IF IT'S STRUCTURAL
OR FUNCTIONAL OR BOTH?
>> SO EVERYTHING THAT WE'VE BEEN
DOING IS LOOKING AT STRUCTURAL
CONNECTOMES, BUT ONE OF OUR
COLLABORATORS IS VERY INTERESTED
IN, SAY, LOOKING AT A STRUCTURE
LIKE THE EYE AND HOW IT HAS
EVOLVED MULTIPLE TIMES OVER
EVOLUTION, SO IF YOU COULD LOOK
AT A CONNECTOME SAY OF A RETINA
FROM A SQUID AND THEN, YOU KNOW,
SOME MORE MAMMALIAN SPECIES AND
COMPARE, YOU MIGHT BE ABLE TO
FIND SOME COMMONALITIES OF THE
CIRCUIT THAT I GUESS TELL YOU
SOME GOOD THINGS ABOUT THAT
DESIGN THAT POP UP EVEN WHEN
THIS HAS EVOLVED INDEPENDENTLY.
YEAH, I'M NOT SURE IF -- DOES
THAT ANSWER YOUR QUESTION?
>> OKAY.
SO I ACTUALLY WANT TO RETURN TO
A QUESTION FOR MOHAMMAD
ACTUALLY.
I JUST FAILED TO WRITE THIS DOWN
MYSELF, BUT I WONDERED IF -- CAN
YOU TELL ME WHAT YOUR SOURCE
DATA IS FOR THAT?
IS IT SOMETHING YOU GENERATED IN
THE LAB SPECIFICALLY OR ARE YOU
USING OTHER DATA?
THERE'S SO MANY REALLY GOOD
DATASETS OUT THERE RIGHT NOW.
OF COURSE MANY OF THEM HAVE NOT
HAD PERTURBATIONS THE WAY YOU
USE BUT I WONDER WHAT THE NEXT
STEP FOR SOME OF THOSE STUDIES
MIGHT BE AND IF YOU CAN USE SOME
OF THE AVAILABLE DATA GENERATED
BY OTHERS.
>> THANK YOU SO MUCH.
THAT'S A GREAT QUESTION.
SO THE DATA, THE ANIMAL
EXPERIENCE, THESE ARE IN VIVO,
THE LTP INDUCTION IS DONE IN
VIVO IN ADULT RAT AND THEN AFTER
THE INDUCTION -- ANIMAL, THEY
ARE TRANSPORTED TO PROFESSOR
KRISTEN -- LAB AT -- AUSTIN FOR
MICROSCOPY AND THEN THE
ELECTROMICROGRAPHS WERE TAKEN,
AND THEN THEY WERE TRANSFERRED
AFTER THAT TO THE LAB FOR THREE
DIMENSIONAL SERIAL SECTION
ELECTROMICROSCOPY SO 3D
RECONSTRUCTION IS DONE IN THE
SIDOSKY LAB, SO AFTER THAT, THE
COMPUTATIONAL ANALYSIS HAS BEEN
DONE AND IT IS VERY IMPORTANT,
THANK YOU FOR POINTING --
REGARDING YOUR NEXT STEPS, SO
THE REALITY OF THESE COULD NECKT
OMIC DATA AND OTHER COLLEAGUES
IN THE GROUP IN THE PANEL HAS
TALKED ABOUT IT IS THAT THEY ARE
RARE AND THEY ARE DONE
DIFFERENTLY.
FOR EXAMPLE, IN OUR DATA, WHICH
IS THE LONG TERM POTENTIATION
INDUCTION OF -- TWO HOUR, THE
THREE HOUR, FOUR HOUR OR ONE DAY
DOES NOT EXIST.
SO THAT'S IT, THAT'S THE DATA,
VERY RARE DATA, THIS IS ONE
LIMITATION.
THE OTHER LIMITATION, MOST OF
THE DATA THAT ARE EXISTING OUT
THERE THAT YOU POINTED, THERE
ARE NOT A LOT, THERE ARE A FEW
CONNECT OMICS DATA, BUT THEY'RE
IN A SINGLE ANIMAL.
SO IF ANYTHING SPECIAL, ANY BIAS
ON THAT ARE THE OR MICE, OKAY,
YOU ARE DONE.
SO THEY CLAIM THEY HAVE, FOR
EXAMPLE, 300 MILLION SYNAPSES,
WHICH IS GREAT, BUT THIS IS FOR
A SINGLE ANIMAL, MICE, RAT OR
DROSOPHILA, SO THIS IS THE
LIMITATION.
I MEAN, WE ARE LUCKY HERE TO
HAVE FIVE RATS.
IT'S NOT A HUGE N, BUT THE
NUMBER OF ANIMALS WERE FIVE.
WE HAD CONTROL, WE HAD LTP30 --
TWO HOURS, SO IT WAS A GREAT
OPPORTUNITY TO INVESTIGATE
DIFFERENT ASPECT OF THE
PRECISIONAL SYNAPTIC -- SYNAPTIC
INFORMATION, STORAGE CAPACITY
AND MANY OTHER ANALYSIS WE HAVE
DONE.
SO I JUST WANTED TO POINT THIS
OUT THAT WE HAVE A VERY LIMITED
DATASET IN COULD NECK TOE MIBG
COMMUNITY TO EXPLORE -- TO
HAVE -- I MEAN, IN OUR FREEDOM
TO EXPLORE DIFFERENT
COMPUTATIONAL METHODS.
YOU NEED TO HAVE A HIGH END TO
DO INFOR INSTANCE ANALYSIS, YOU
NEED TO HAVE DIFFERENT
CONDITIONS TO FIND CORRELATION
TO FIND CAUSATION, AND WE ARE
VERY LIMITED TO THE DATA, TO THE
QUALITY.
I AS A COMPUTATIONAL PERSON HAD
THE FOREFRONT OF THE PROJECT
NEEDS MANY DIFFERENT CHANGES BUT
THERE IS NO WAY TO DO THOSE
CHANGES BECAUSE WE SHOULD GO
BACK AND DO THE EXPERIMENT.
SO THEY FIRST DO THE EXPERIMENT
AND THEN AT THE LAST STAGE, THE
COMPUTATIONAL PERSON STARTS
ANALYZING AND ANALYZING LATER
AND YOU NEED MANY THINGS BUT YOU
CAN GO BACK AND DO ANOTHER
ANIMAL, BUT REGARDING THE OTHER
QUESTIONS YOU HAD, THESE ARE
HAND TRACED, SO THE DATA WE
HAVE, THE ACTUAL MICROGRAPHS
THAT ARE PROVIDED AT AUSTIN,
THEY WERE MAYBE TWO YEARS OF
HAND TRACING TO HAVE THESE
MICROGRAPHS READY FOR 3D
RECONSTRUCTION.
SO I HOPE I ADDRESSED ALL THE
QUESTIONS YOU POINTED.
>> YEAH, THANK YOU.
THOSE CLARIFICATIONS ARE REALLY
A VERY SIGNIFICANT POINT.
THANK YOU.
>> THANK YOU.
>> BENJAMIN, YOU HAVE A HAND UP.
WOULD YOU LIKE TO JUMP IN?
>> I REALLY WOULD LIKE TO JUST
ECHO WHAT MOHAMMAD WAS SAYING
ABOUT THE INCREASING NEED IN
COULD NECK TOE MIBGS TO HAVE
MULTIPLE SAMPLES WHERE YOU CAN
START TO DO PER TEU BAIGS AS
WELL PERTURBATIONS AS
WELL AS SORT OF HAVING AN IDEA
OF WHAT WE'RE LOOKING FOR BEFORE
WE CONNECT THE CONNECTOME AND
SORT OF WHAT ANALYSIS ONE COULD
DO TO GET OUT THESE QUESTIONS
THAT WE'RE INTERESTED IN, JUST
SOMETHING THAT'S COME UP FOR ME
AS WELL IN MY WORK, SO I JUST
ANY TIME WAKED TO ECHO W.
>> LET ME BUILD ON THAT SLIGHTLY
AND MAYBE TAKE A BIT OF A PIVOT.
THE NIH HAS BEEN INVESTING IN
SORT OF THIS CONNECTOME SPACE
FOR A WHILE.
IT HAS GROWN CONSIDERABLY AND IT
HAS CHANGED IN CIRCUIT -- CROSS
LEVELS AS WELL.
AND THERE HAS BEEN A BIG PUSH
AND SO MOHAMMAD SORT OF TOUCHED
ON THIS, THE IDEA OF SLAIRED
DATASETS.
HOW VALUABLE ARE THEY, WHAT'S
THE VARIABILITY BETWEEN THEM,
WHAT'S CONSISTENT BETWEEN THEM
SO YOU CAN ACTUALLY POOL THINGS.
THAT'S BEEN A MASSIVE PUSH.
WE'VE HEARD A LOT OF CIRCUIT
STUFF BUT THIS GOES DOWN TO THE
CELLULAR LEVEL AS WELL.
CERTAINLY AT LEAST MOHAMMAD
YOU'RE TALKING ABOUT CHRISTIAN
HARRIS' WORK, WE'RE TALKING
ABOUT EM LEVEL AS WELL.
SO THAT'S A CHALLENGE.
THAT'S ONE OF THESE EVOLVING
CHALLENGES, THE IDEA THAT THE
NIH IS PUSHING ON DATA SHARING
SO THAT YOU HAVE THESE RESOURCES
AVAILABLE TO MAXIMIZE THE
INVESTMENT THAT THE TAXICABS
PAIR HAS MADE TO TS
MADE IN SCIENCE.
TO PIVOT SLIGHTLY, THOUGH, I'M
NOT GOING TO PUT YOU BY NAME ON
THE SPOT BUT I'M GOING TO ASK
THOSE PEOPLE WHO ARE MORE SENIOR
ON THE PAM, PANEL, SENIOR, AE
TERM, RIGHT?
THOSE OF YOU THAT ARE POSTDOCS,
YOU'RE THE SENIOR PEOPLE ON THIS
PANEL, OKAY?
SO THOSE OF YOU THAT ARE
POSTDOC, I'M WONDERING IF YOU
COULD SHARE YOUR THOUGHTS AS TO
HOW YOU SELECTED YOUR POSTDOC
LAB, WHAT SOME OF THOSE
CHALLENGES HAVE BEEN AND WHAT
RECOMMENDATIONS YOU MIGHT GIVE
TO YOUR OTHER SELF IF YOU CAN GO
BACK IN TIME.
OBVIOUSLY YOU DON'T WANT TO SAY
ANYTHING THAT'S GOING TO PUT NEW
HOT WATER, BUT I KNOW FOR SOME
OF YOU, AT LEAST ONE OF YOU,
YOU'VE JUMPED INTO THAT LAB THAT
WAS BRAND NEW ESSENTIALLY, AND
THERE ARE UNIQUE CHALLENGES WITH
THAT.
SO IF YOU HAVE THOUGHTS ABOUT
THAT, I THINK THERE MAY BE
OTHERS THAT ARE LISTENING WHO
WOULD POTENTIALLY BENEFIT FROM
THAT.
EVERYBODY IS TRYING TO SAY
SOMETHING, YOU'RE ALL ON MUTE.
GO AHEAD AND UNMUTE.
>> SO I THINK I'M KIND OF SENIOR
HERE.
SO I ACTUALLY STAYED IN MY OWN
LAB.
I DID MY PH.D. WITH -- AND
STAYED HERE BECAUSE -- GIVE US
THIS GREAT OPPORTUNITY TO --
WHICH ALLOWS US TO PROVIDE
SERVICE TO THE GREAT
NEUROSCIENCE COMMUNITY.
THE OTHER REASON I STATE HERE IS
DEFINITELY BECAUSE THIS PROJECT
THAT'S DEVELOPING POLYMER
DEVICES, FOR MYSELF I THINK ONE
BIGGEST CHALLENGE WITHIN THIS
NEUROSCIENCE AND
NEUROENGINEERING COMMUNITY IS
STILL HOW TO COLLECT VERY HIGH
QUALITY LONG TERM -- DATA.
I THINK THAT'S THE FOUNDATION TO
UNDERSTAND ALL THE FUNCTIONS OF
OUR BRAIN.
SO IF WE CAN DEVELOP TOURS TO TO
BETTER COLLECT -- TO DO THIS
FOUNDATION, I THINK IT'S VERY
MEANINGFUL.
SO MY EXPERIENCE DOESN'T -- I
KNOW IT'S LIKE NOT -- SOME
PEOPLE SAY IT'S NOT GOOD IF YOU
WANT TO STAY IN ACADEMIA, IT'S
NOT GOOD TO STAY IN THE SAME LAB
FOR VERY LONG.
BUT MINE IS A VERY SPECIAL CASE.
SO JUST WANT TO START TO -- SO
PEOPLE CAN JUMP IN.
>> SO LET ME PUSH ON THAT A
LITTLE FURTHER.
IN FACT I THINK -- ROHIT, YOU
HAVE A QUESTION THAT SORT OF
BUILDS ON THAT AS FAR AS
ALIGNING -- NOT ALIGNING BUT
SORT OF THINKING ABOUT YOUR
RESEARCH DIRECTION AND HOW IT'S
GOING TO BE SUPPORTED IN THE
FUTURE.
DO YOU WANT TO ASK YOUR
QUESTION?
>> YES, SURELY.
SO MAYBE NOT -- THIRD YEAR
GRADUATE STUDENT, I'M SLOWLY
THINKING THERE'S QUESTIONS I
WANT TO ASK FURTHER PROBABLY
AFTER MY PH.D., BUT -- AND I
WANT TO DO A -- IN ACADEMIA.
AND THEN I'M THINK HOW SHOULD I
NARROW THIS DOWN IN TERMS OF
PRIORITY SO THAT I CAN ALSO GET
EASY FUNDING FROM THE FUNDING
AGENCIES.
LIKE HOWL TO DO HOW TO DOO
START DOING THAT, WHEN TO START
APPLYING FOR GRANTS IS A
QUESTION OF THAT.
>> SO I SUPPOSE FROM A
MANAGEMENT PERSPECTIVE, I'M IN
PROBABLY THE BEST POSITION TO
ANSWER THE QUESTION, BUT I WANT
TO SEE IF ANYBODY ON THE PANEL
HAS ANY THOUGHTS FIRST.
>> OR IF YOU'VE GOTTEN ANY
ADVICE ALONG THE LINES FROM YOUR
MENTORS OR ADVISORS, OR IF YOU
WANT TO THINK ABOUT IT FOR A
SECOND, JAY CAN GIVE THE
MANAGEMENT EXPERIENCE THAT HE
HAS.
>> YEAH, SO LET ME SORT OF MAYBE
BREAK THE ICE ON THAT.
ONE IS, ACTUALLY I WILL HAVE A
LITTLE BIT BECAUSE YOU SAID THE
EASY PATH TO FUNDING, WHAT IS
THAT?
THAT DOES NOT EXIST.
AND THE SOONER WE CAN ACCEPT
THAT, THAT THAT IS A REALITY,
THE BETTER OFF WE'LL BE.
UNFORTUNATELY THERE IS MORE
GREAT SCIENTIFIC QUESTIONS THAN
THERE ARE MONEY TO SUPPORT IT,
IS WHAT IT COMES DOWN TO.
SO HOW DO YOU MOVE FORWARD?
I THINK THERE'S A COUPLE OF
WAYS.
ONE IS THAT I THINK YOU FIRST OF
ALL FROM A HYPOTHETICAL
PERSPECTIVE, YOU WANT TO LOOK TO
SEE WHAT'S WORKED FOR OTHERS.
THIS IS CHALLENGING.
WE HAD INITIAL CONVERSATIONS
ABOUT HOW DOES SOMEBODY COME AT
THIS FROM AN ENGINEERING SPER
SPECULATIVE WHERE THE NIH IS A
BIT OF A FOREIGN ENTITY.
A LOT OF ENGINEERS, NOT ALL BUT
A LOT OF ENGINEERS, THIS IS NOT
THEIR NORMAL SORT OF WAY.
I WILL SAY THE BRAIN INITIATIVE
HAS REALLY PUSHED EXPANDING THE
DIVERSITY OF PERSPECTIVES WE
HAVE, AND THAT DIVERSITY SPANS A
LOT OF DIFFERENT DIMENSIONS.
ONE OF THOSE IS ACTUALLY CAREER
EXPERTISE.
SO WE WANT TO BRING IN PEOPLE
WHO ARE OUTSIDE OF OUR NORMAL
SHALL WE SAY CHOIR OF
INVESTIGATORS.
NOT JUST NEUROSCIENTISTS FOR THE
BRAIN INITIATIVE, BUT PEOPLE WHO
REALLY BRING A DIFFERENT
PERSPECTIVE.
SO WE VALUE -- I MEAN, I LOOK
AROUND THIS VIRTUAL ROOM IN
FRONT OF US.
WE VALUE THE DIFFERENT
PERSPECTIVES YOU HAVE.
SO IT IS HUGELY IMPORTANT THAT
WE FIND ANSWERS FOR THAT.
NOW THE MORE DIRECT ANSWER, I
WOULD SAY, IS THAT THERE ARE
FUNDING MECHANISMS AVAILABLE TO
ALL CAREER STAGES FOR A VARIETY
OF DIFFERENT REASONS.
NOW I'LL BE THE FIRST TO ADMIT,
YOU'RE GOING TO LOOK AT THESE
LETTERS, THESE Ks AND Fs AND
ALL OF THESE THINGS AND IT'S
GOING TO BE MUMBO JUMBO TO A LOT
OF YOU.
BOUTS BECAUSE IT'S A FOREIGN .
BR THERE'S WHERE NIH PROGRAM
OFFICIALS ARE HERE TO HELP.
SO ANDREA AND I, THE TWO THAT
ARE ON THIS CALL RIGHT NOW, CAN
ABSOLUTELY HELP OUT.
IF IT'S NOT RELEVANT FOR MY
RESEARCH PROGRAM, I DON'T WANT
TO JUST SAY NO THANK YOU AND
PUSH YOU OFF.
WE WANT TO FIND THE RIGHT PLACE.
AND SOMETIMES THE RIGHT PLACE
ISN'T OH, WE HAVE A PROGRAM FOR
THAT.
SOMETIMES IT'S, SORRY, THAT'S
ACTUALLY NOT IN OUR LANE FOR
WHAT WE CAN DO.
HAVE YOU THOUGHT ABOUT THESE
OTHER PLACES AS WELL?
SO REALLY THE LONG WINDED SORT
OF SUM REAL THERE IS REALLY --
IS REACH OUT TO PROGRAM
OFFICIALS, SEND US AN EMAIL,
SEND US YOUR SPECIFIC AIMS IF
YOU HAVE THOUGHTS IN MIND, IN
GENERAL, A DRAFT OF THAT, YOUR
BIOSKETCH, AND WE ARE HERE TO
HELP.
>> I JUST WANT TO REINFORCE THAT
FOR EVERYBODY ON THIS CALL AND
ANYBODY WHO MIGHT BE LISTENING,
PROGRAM OFFICERS AT THE NIH
EXIST TO TALK TO APPLICANTS AND
GRANTEES.
THAT IS WHAT OUR JOB IS.
SO YOU SHOULD FEEL FREE TO REACH
OUT EARLY AND OFTEN AND THAT IS
WHAT OUR TRAINING EXPERTS DO AND
THEN WHEN YOU GET FURTHER INTO
YOUR CAREER WHEN YOU'RE LOOKING
FOR THE R MECHANISM, THE R01s
AND THE R21s ABANDONS AND ,
PROGRAM OFFICERS LITERALLY EXIST
TO HELP PEOPLE GET FUNDED, TO
UNDERSTAND THE SITUATION, TO
NAVIGATE THE BIG BLACK BOX
THAT'S NIH, AND IT'S WHAT WE
ENJOY DOING AND WE LOVE TO TALK
SCIENCE WITH YOU.
WE JUST NEED YOU TO REACH OUT TO
US.
AND BE PERSISTENT, BE PATIENT.
YOU MAY ALSO GET FORWARDED, AS
JAY SAID, IF HE'S NOT THE THE
RIGHT TARGET, HE WILL SEND TO
YOU A COLLEAGUE THAT HE THINKS
CAN HELP YOU BE BETTER
POSITIONED.
SO THAT IS A FUNDAMENTAL LESSON
THAT EVERYBODY SHOULD LEARN NO
MATTER WHAT CAREER STAGES, TO
REACH OUT TO PROGRAM OFFICERS
ACROSS THE DIFFERENT INSTITUTES
AND ALLOW THEM TO ASSIST YOU.
HOW ARE WE DOING FOR TIME?
I KNOW WE'RE WRAPPING UP IN JUST
A LITTLE BIT.
THERE WERE A COUPLE MORE
QUESTIONS IN THE CHAT, JAY.
WE HAVE A 2-MINUTE WARNING.
IF WE CAN GET TO THOSE SO THAT
THOSE INDIVIDUALS GET THEIR
QUESTIONS ANSWERED, SO I'LL DO
THE FIRST ONE THAT IS TO
DR. PEDEGO AGAIN.
WHAT METHOD ARE YOU USING TO
FIND THE, QUOTE-UNQUOTE,
NETWORKS?
AND WHAT DATA ARE -- SORRY, AND
WHAT DATA PROVIDE THE STARTING
POINT?
SO IF YOU HAVE A QUICK ANSWER TO
THAT, THEN WE'LL GET TO THE LAST
QUESTION WE HAVE.
>> YEAH, VERY QUICKLY, IT'S
ELECTRON MICROSCOPY DATA, SO
IT'S PURELY BASED ON THE
STRUCTURE AND SYNAPSES THAT
PEOPLE OBSERVE IN THOSE IMAGES
AND THEN THE RECONSTRUCTION IS
DONE VIA MANUAL TRACING.
>> GREAT, THANKS.
MAYBE AS A GOOD BOOK END,
BECAUSE I BELIEVE THE FIRST
QUESTION WAS FOR DESIREE, WE'RE
GOING TO SEND THE LAST QUESTION
AS WELL.
THERE'S A COUPLE PIECES TO THIS.
I'LL TUMP ON CAN YOU COMMEF
INACTIVATIONS ON THE PHOTO
ACTIVATED OXYMORPHINE OR ANY
OTHER LONG-ACTING OR EXTENDED
RELEASE MEDICATIONS THAT YOU
HAVE USED?
>> YEAH, SO QUICK ANSWER TO
THIS, CURRENTLY WITH THE IN VIVO
DATA THAT IS SHOWN LIKE ON MY
POSTER, YOU SEE THAT BOX, PHOTO
ACTIVATION LASTS TENS OF MINUTES
IN OUR IN VIVO DATA, BUT IN THE
FUTURE, WE'RE TRYING TO OPTIMIZE
OUR DOSE CONCENTRATION LIGHT
USAGE SO THAT WE CAN HAVE, YOU
KNOW, QUICKER OFFERING
ESSENTIALLY TO LOOK AT, JUST
MORE SENSITIVE BEHAVIORS AND
NEUROMECHANISMS.
>> GREAT.
SHALL I TAKE A STAB, JAY, REAL
QUICK?
>> PLEASE.
>> I WANT TO THANK EVERYBODY FOR
THEIR TALKS TODAY.
IT WAS REALLY GREAT.
WE CAN'T WAIT TO SEE WHAT YOU
BRING TO US IN THE FUTURE AND
WHAT YOUR FUTURE HOLDS FOR ALL
OF YOU.
WE APPRECIATE IT, EVERYBODY GO
CHECK OUT THE POSTERS, AND WE
LOOK FORWARD TO THE REST OF THE
MEETING.
THANK YOU.
>>HELLO, AND WELCOME TO THE
SYMPOSIUM, ADVANCING OUR
UNDERSTANDING OF THE BRAIN AND
BEHAVIOR THROUGH CONNECTOMICS.
TODAY WE HAVE SIX TALKS AND I'LL
BRIEFLY INTRODUCE THE SPEAKERS
TODAY.
BEN MULCAHY FROM THE LONDON FELD
RESEARCH INSTITUTED IN TORONTO,
SPEAKING NEXT WILL BE CLAIRE
MCKELLAR FROM FLY WAIR, I WILL
BE FOLLOWING CLAIRE, MY NAME IS
WEI-CHUNG LEE BOSTON CHILDREN'S
HOSPITAL AND HARVARD MEDICAL
SCHOOL.
FOLLOWING ME WILL BE STEPHANIE
HAMPEL FROM THE UNIVERSITY OF
PUERTO RICO AND FOLLOWING
STEPHANIE WILL BE MICHAEL REISER
FROM HOWARD HUGHES MEDICAL
INSTITUTE -- RESEARCH CAMPUS.
FINALLY, WE'LL HAVE SETH GRANT
FROM EDENBUROUGH UNIVERSITY AND
FOLLOWING SIX TALKS WE'LL HAVE
TEN TO FIFTEEN MINUTE
QUESTION-AND-ANSWER SESSION WITH
MOST OF THE SPEAKERS.
SO WE WILL START WITH BEN.
>> GREAT.
THANKS TO THE ORGANIZERS FOR
INVITING ME TO SPEAK TODAY.
I'M BASED IN TORONTO AND FOR THE
LAST FEW YEARS I'VE BEEN TRYING
TO UNDERSTAND HOW NEURO CIRCUITS
MODULE AFTER US, SO THESE
MODELING CIRCUITS ARE WIDESPREAD
ACROSS, THEY INCLUDE THE BIRTH
OF NEW NEURONS WHICH HAVE TO GO
THROUGH PROCESSES AND INTEGRATE
INTO EXISTING CIRCUITRY AND
JUVENILE CIRCUITS THAT HAVE TO
REWIRE TO ACCOMMODATE NEURONS OR
MODIFY THEIR PROPERTIES AND AS
THIS REMODELING TAKES PLACE,
CIRCUITS STILL HAVE TO BE ABLE
TO WORK WITHOUT BEING DISRUPTED.
C HE WILL GAFNS DOES THIS VERY
WELL AND REHATCHES THROUGH 220
NEURONS AND INCREASES THIS
NUMBER TO 302.
MOST OF THESE NEW NEURONS ARE
MOARTD NEURONS EXPANDING THE
MOLARITY NEURON POOL FROM 22 TO
75 METANEURONS AS WELL AS
INCREASING THE NUMBER OF
METANEURON CROSSES.
IN THE FIRST STAGE C. ELEGANS
HAS THIS ASYMMETRIC, HAS TWO
CLASSES OF NEURONS TO CONTRACT
DORSAL MUSCLES AND THIS ALSO
ACTIVATES THESE INHIBITOR
NEURONS WHICH SIGNAL ALONG THE
PROCESS TO REALLY SCAMPER TO
INHIBIT VENTRALS, SO YOU HAVE
THIS EXCITATION OF DORSAL
MUSCLE, RELAXATION OF VENTRAL SO
YOU GET BENDING.
9 SICIALGHT HAS EXPANDED WITH
ALL OF THESE NEW CAUSES OF
METANEURONS AND IT'S BECOME
SYMMETRIC AND THIS ISN'T JUST
DUE TOLT ADDITION OF
FUNCTIONALITY BY THE NEW NEURONS
BUT ALSO REMODELING OF THE
JUVENILE METANEURONS, SO THE
METANEURONS THAT INHIBITED
METANUFLS IN JUVENILE ANIMALS
SWAWL POLAR TAKE AND JUVENILE
ROLE IS TAKEN OVER BY THESE
NEWBORN INHIBITORY NEURONS WHICH
GROW OUT AND COPY THEIR WIRING
PATTERN.
REMARKABLY THIS HAPPENS WITHOUT
DISRUPTING CIRCUIT OUTPUT.
C. ELEGANS IS ABLE TO MOVE
AROUND HAPPILY BEFORE, DURING
AND AFTER MOTOR REMODELING.
SO IF WE TRACK ANIMALS MOVING
AROUND THE PLATE AND EXPRESS G
CAMP AND MUSCLE CELLS WE CAN SEE
THEY MOVE AROUND WELL WITH BODY
BENDING WAVES THAT TRAVEL FROM
THE HEAD TO THE TAIL, ON THE
BODY, AND VENTRAL AND DORSAL
CALCIUM WAVES OF THE MUSCLES
THAT CORRELATE WITH THE BENDING
NICELY.
VENTRAL AND DORSAL CALCIUM ARE
ANTI-CORRELATED, THIS IS THE
CASE AT THE BEGINNING, MIDDLE
AND END OF MODELING AS YOU CAN
SEE FROM THIS HISTOGRAM WHERE
VENTRAL AND DORSAL CALCIUM
ACTIVITY IN RED AND BLUE IS
180 DEGREES OU OF PHASE, BUT
180 DEGREES, SO TO UNDERSTAND
HOW THIS CIRCUIT REMODELS
WITHOUT DISRUPTING MOTOR ABILITY
WE DECIDED TO LOOK AT THE OLDER
STRUCTURE OF THE METACIRCUIT AS
IT REMODELS.
TO DO THIS, WE DID ZEER --
ELECTRON MIE CROSS SWOPE BEFORE,
DURING AND AFTER REMODELING AND
FIRST COVERED THE WHOLE BODY,
NOSE TO TAIL, THEN WE REALIZED
THIS IS ACTUAL A LOT OF WORK AND
EVERYTHING IS REPEATED ALONG THE
BODY ANALYSIS ANYWAY SO THE LAST
TWO RECONSTRUCTIONS FOCUS ODD
THEIR ANTERIOR BODY AND INCLUDED
THE ENTIRE BODY DD1 AND 2 AND
BECAUSE THE DEVELOPMENTAL
TIMELINE AND C. ELEGANS
STEREOTYPES THIS MEANS THAT BY
LOOKING AT DIFFERENT INDIVIDUALS
AT SEQUENTIAL DEVELOPMENTAL
STAGES WE ESSENTIALLY END UP
WITH A TIME MAX OF REMODELING
ACROSS DEVELOPMENT AND NOT ONLY
THAT BUT YOU MIGHT SEE FROM THIS
RECONSTRUCTION OF THE NEW
INHIBITORY THAT MIDWAY THROUGH
OUTGROWTH THE ANTERIOR NEURONS
ARE MORE ADVANCED STAGE OF OUTER
GROWTH THAN THE POSTERIOR ONES
AND THIS FOLLOWS THE BIRTH ORDER
OF THE NEURONS SO WE HAVE THIS
TIME LAPSE BETWEEN INDIVIDUALS
AND WITHIN INDIVIDUALS.
THE FIRST THING WE SAW WHEN WE
LOOKED AT THE DATA WAS THAT
PROCESS OUTGROWTH WAS VERY
INTERACTIVE SO AT THIS TIME
POINT IN THE FIRST LARVA STAGE
WE HAVE CAUGHT THIS NEWBORN
INHIBITORY MOTOR NEURON HALFWAY
THROUGH THE OUTGROWTH OF ITS
PROCESS.
IF YOU TAKE A CROSS-SECTION
THROUGH THIS COMMISSION WHAT YOU
SEE IS THE NEWBORN NEURONS
WRAPPING AROUND THIS EXISTING
COMMISSION BUNDLE.
THEN IF YOU LOOK ONE HOUR LATER
AFTER NEURONS REACH THE DORSAL
COURT, YOU SEE IT'S NOW THE
DORSAL PROCESS THAT IS WRAPPING
AROUND EXISTING NEURONS
ESPECIALLY JUVENILE INHIBITORY
MOTOR NEURONS, AND FINALLY AFTER
OUTGROWTH IS COMPLETE, YOU SEE
THE NEURO NEURONS ARE RUNNING IN
PARALLEL AND YOU HAVE THIS
MORPHOLOGY THAT ISN'T WRAPPING
AROUND EXISTING STRUCTURES
ANYMORE.
SO WE HAVE THIS SEQUENTIAL
WRAPPING AND UNWRAPPING AROUND
THE INHIBITORY MOTOR NEURONS
THAT OCCURS DURING JOAT GROWTH
LIKELY HELPING TO GUIDE THE
NEWBORN NEURONS TO PROCESS
PLACEMENT.
IF YOU LOOK AT THEIR
CONNECTIVITY YOU SEE THE NEWBORN
NEURONS START TO FIRE UP DURING
OUTGROWTH.
SO HERE WE HAVE RECONSTRUCTION
OF THE B -- VD3 NEURON HALFWAY
THROUGH THE OUTGROWTH.
MATURING NEURONS ON LEFT,
RECEIVES INPUTTED AND NEW
MUSCULAR JUNCTIONS BUT DOESN'T
HAVE ANY INPUTTED F WE TAKE A
CROSS-SECTION THROUGH THE
VENTRAL PROCESS WE SEE A SYNAPSE
IN THE PROCESS OF ESTABLISHING
ICHTS THAT HAS A PRESYNAPTIC
PROJECTION POINTING TOWARDS
MUSCLE CELL AND IT DOESN'T YET
HAVE AN ESTABLISHED BASE CORE
COWBTD AND AS TIME GOES ON,
RECRUITED AND SYNAPSE BECOMES
MATURE.
SO SYNAPSE GENESIS OCCURS DURING
OUTGROWTH OF THESE NEW NEURONS
AND IT'S VERY LIKELY A HARD
HARDWIRED PROCESS THAT BEGINS
WITHOUT ANY VENTRICULAR
COMPONENT.
NOW F WE LOOK AT THE CONNECTED
ACTIVITY OF THE JUVENILE MOTOR
NEURONS WE SEE THAT EARLY IN THE
FIRST LARVAL STAGE AS EXPECTED
THEY RECEIVED DORSAL INPUT AND
MAKE DORSAL NEW JUNCTIONS.
SAYS AFTER REMODELING THIS
PATTERN IS REVERSED, THEY
RECEIVE VENTRAL INPUT AND MAKE
NEW MUSCULAR JUNCTIONS TO DORSAL
MUSCLE CELLS.
IN THE MIDDLE OF THIS REMODELING
THEIR POLARITY IS MIXED, THEY
GET INPUT ON BOTH THE VENTRAL
AND DORSAL SIDES AND MAKE NEW
MUSCULAR JUNCTIONS TO BOTH SIDES
TOO.
ONE MIGHT EXPECT THAT THIS WOULD
DISRUPTED CIRCUITRY IN THE MOTOR
BUT DON'T SEE THIS DISRUPTION IN
THE BEHAVIOR.
WE LOOK AT CLUSTERING IN MUSCLE
CELLS AND EARLY IN DEVELOPMENT
WE SEE THEM ONLY ON THE VENTRAL
SIDE OF THE ANIMAL CONSISTENT
WITH THE CONNECTIVITY AND AFTER
REMODELING WE SEE THEM ON BOTH
VENTRAL AND DORSAL SIDES.
AND WHEN WE LOOK DURING MODELING
WE START TO SEE THE APPEARANCE
OF THESE FAINT CLUSTERS ABOUT
THE SAME KIND OF TIME AS WE
START TO SEE SYNAPSES APPEARING
IN THE ELECTROMICROSCOPY POINTS,
THEY'RE WEAK AND THEY APPEAR IN
THE ANTERIOR BODY FIRST AND THEN
THEY PROGRESS ALONG THE
POSTERIOR.
SO WE HAVE THIS JUVENILE MOTOR
CIRCUIT NEW NEURONS AND
REMODELING EXISTING ONES AS IT
SWITCHES FROM JUVENILE TO MATURE
CONFIGURATION.
DURING THE REMODELING WE SEE
THIS GRADUAL BUILDUP OF THE
MATURE CIRCUITED TRI WHILE
MAINTAINING THE JUVENILE
CONNECTIONS AND IT'S ONLY AFTER
EVERYTHING IS BUILT UP THAT THE
JUVENILE CONNECTIONS ARE
DISASSEMBLED AND THE CIRCUIT
SWICHEDZ FROM THE JUVENILE TO
MATURE INTEGRATION.
THIS IS STAGGERED ALONG THE
ANTERIOR POSTERIOR ACCESS
FOLLOWING THE BIRTH ORDER OF THE
NEURONS.
SO TOGETHER THIS GRADUAL BUILDUP
OF NEW CONNECTIVITY AND THE
SWITCH ALONG WITH STAGGERED
REMODELING LOONG THE BODY AXIS
LIKELY HELPS ONE TO REMODEL ILTS
MOTOR CIRCUIT WITHOUT DISRUPTING
MOTOR COMMISSION.
LASTLY THIS SYSTEM TAKES A VERY
DIFFERENT APPROACH TO MATURATION
COMPARED TO CENTRAL NERVOUS
SYSTEM.
DANIEL IN OUR LAB SHOWED THAT
CNS LARGELY BUILT BY BIRTH AND
MAINTAINED ACROSS MATURATION
WITH WIRING CHANGES HAPPENING
ALONG THIS CONSTANT SCAFFOLD.
THE BIRTH SYSTEM UNDERGOES THIS
WIDE SCALE REMODELING OF ITS
STRUCTURE, WIRING AND ITS
COMPLEMENTED NEURONS, SO
DIFFERENT PARTS OF THE NERVOUS
SAM CONTAIN VERY DIFFERENT
APPROACHES TO REMODELING AND IF
YOU WOULD LIKE TO READ MORE
ABOUT THIS STORY, IT'S UP ON THE
BIORX PAGE.
THANKS FOR EVERYBODY INVOLVED,
AND TO THE NIH AND C HFR FOR
FUNGSD.
>> I'M CLAIRE MCKELLAR DIRECTOR
OF OPERATIONS AND COMMUNITY FOR
FLY WIRE OPEN SCIENCE PROJECT AT
PRINCETON.
FLY WIRE IS A HUMAN AI
COLLABORATION TO MAP THE FLY
CONNECT TOMORROW, PI'S ARE IS HE
BASS STAN AND MALLA MUR MURPHY.
SOME OF THE LARGER PROJECTS IN
THE FLY BRAIN THE LARVA WHERE IT
MANUALLY RECONSTRUCTED CONNECT
TOMORROW OF THE WHOLE --
CONNECTOME, HEMIBRAIN, PART OF
ADULT CENTRAL BRAIN, IMAGE
DATASET OF FULL FEMALE ADULT FLY
BRAIN AVAILABLE?
IS WALLED GARDEN ENVIRONMENT
WHERE MEMBERS CAN MANUALLY
RECONSTRUCT NEURONS AND FLY WIRE
USES THE SAME IMAGE FLY SET BUT
NEURONS RECONSTRUCTED BY OUR AI
AND PROOFREAD BY AN OPEN
COMMUNITY OF FLY BIOLOGISTS AND
OTHERS.
THE GOAL OF FLY WIRE IS TO
CROWDSOURCE A WHOLE BRAIN
CONNECTOME FOR THE ADULT.
USED TRANSMISSION ELECTRON
MICROSCOPE ON AS STACK OF 7,000
SECTIONS, DATA SET OF 21 MILLION
CAMERA IMAGES.
AT PRINCE DONE WE LINED NEW
ALIGNMENT AND SEGMENTATION
PIPELINE TO IMPROVE HOW NEURONS
COULD BE RECONSTRUCTED BY
ARTIFICIAL INTELLIGENCE AND RAN
THIS ON THE WHOLE BRAIN.
THE AI DETECTS SEGMENTS THAT ARE
OFTEN RECOGNIZABLE AS NOO
NEURONS, FOR EXAMPLE THESE GIANT
FIBER NEURONS INVOLVED IN HE
INVOLVED IN ESCAPE CIRCUITED
COMPARED TO ONES FROM PRINCETON
AI TO LIGHT MICROSCOPYING
STAINING IN THE LITERATURE AND
NEURON INVOLVED IN MEMORY.
THE AI IS VERY GOOD BUT NOT
PERFECT, MAKING ERRORS THAT
REQUIRE HUMAN PROOFREADING.
HERE IS A NEURON THAT GOT BROKEN
BECAUSE OF A MERGER WITH A --
CELL.
HERE ARE TWO NEURONS
ACCIDENTALLY MERGED, ONE
INVAGINATED INTO INTO ANOTHER.
HERE IS ONE THAT GOT BROKEN OFF
FROM WHERE IT LEFT OFF A LARGER
BACKBONE, HERE ARE A BUNCH OF
TWIGS, VERY FINE LITTLE
PROCESSES AT THE ENDS OF
DENDRITES THAT GOT ORPHANED OR
BROKE BROKEN OFF THE END, THAT
ACTUALLY HAPPENS WITH SOME
FREQUENCY.
BUT WE FIND THAT PROOFREADING
ORPHANED TWIGS SNTD CRITICAL --
IS NOT CRITICAL BECAUSE ANY TWO
CONNECTED NEURONS TEND TO BE
REDUNDANTLY CONNECTED.
SO EVEN IF SOME TWIGS ARE
MISSIONING, IT DOESN'T SEEM TO
AFFECT THE WIRING DIAGRAM.
SO WE MAINLY JUST PROOFREAD
BACKBONES OR THE LARGER BRANCHES
OF NEURONS AND NOT THE FINE
TWIGS.
BUT EVERYTHING ELSE NEEDS HUMAN
INTERVENTION.
SO WE CREATED FLY WIRE AS AN
ONLINE PLATFORM TO CROWDSOURCE
THE PROOFREADING THIS THIS WHOLE
BRAIN DATASET.
ANY SIGN SCIENTIST CAN JOIN TO
HELP PROOFREADS READ INTERESTS,
USERS NEED TO PASS A
PROOFREADING TEST ON ENTER THE
COMMUNITY.
FLY WIRE'S USER INTERFACE IS
BAIS ODD GOOGLE'S NEURO GLANCER
WITH OUR OWN MODIFICATIONS.
BEHIND UI IS OUR SPATIALLY
CHUNKED SUPER GRAPH THAT ALLOWS
SIMULTANEOUS COLLABORATIVE
EDITING.
USESSERS AGREE TO OUR PRINCIPLES
OF OPENNESS, FAIRNESS,
COMMUNICATION AND MUTUAL
RESPECT, WHICH ENSURE THAT
PEOPLE RECEIVE CREDIT FOR THEIR
WORK.
THESE PRINCIPLES WERE DESIGNED
WITH INPUT FROM OUR ADVISORY
BOARD AND IT'S BEEN GRATIFYING
TO SEE THAT SCIENTISTS ARE
SHARING THEIR DATA VERY OPENLY
BECAUSE OF THIS CAREFUL SYSTEM
OF CREDIT.
I HAVE NOW TALKED TO HUNDREDS OF
MEMBERS OF THE FLY COMMUNITY AS
I ONBOARD THEM AND I'M HUMBLED
AND APLAITZED BY -- AMAZED BY
HOW CAN READY PEOPLE ARE FOR
OPEN SCIENCE.
WE HAVE FLY WIRE MEMBERS FROM
MANY INSTITUTIONS ALL AROUND THE
WORLD.
PROOFREADING NEURONS IN FLY WIRE
IS REALLY FUN AND KIND OF
ADDICTIVE LIKE A VIDEO GAME
BECAUSE OF THE 3D EXPERIENCE.
YOU JUST DROP POINTS ON OBJECTS
THAT YOU WANT TO SPLIT APART,
FOR EXAMPLE, HERE IS A NEURON,
THE TUBULAR THING, THAT GOT
ACCIDENTALLY MERGED BY THE AI
WITH A BUNCH OF GLIA WHICH KIND
OF LOOKS LIKE SEAWEED AND JAMES
IS DROPPING RED POINTS ALONG THE
NEURON AND BLUE POINTS ALONG THE
GLIA TO SPLIT THEM APART.
AND THIS SUCCESSFULLY CUT THOSE
TWO THINGS, THE OUTER IS VERY
SMART -- THE ALGORITHM IS VERY
SMART SO YOU CAN BE KIND OF
ROUGH WITH YOUR POINT PLACEMENT.
THEN ONCE JAMES HIDES THE GLIA
CELL YOU CAN SEE THIS CLEANED UP
NEURON.
MERGING BROKEN PIECES IS EVEN
EASIER THAN SPLITTING, SO IF A
CELL WAS MISSING PART OF ITSELF
ONCE YOU FIND THE CONTINUATION
IT JUST TAKES TWO CLICKS TO ADD
THAT PIECE.
PUTTING TOGETHER CIRCUITS
REQUIRES KNOWING WHERE SYNAPSES
ONTO ONE ANOTHER, SO WE IMPORTED
SYNAPSES AUTOMATICALLY DEFECTED
BY THE LAB AND THIS CELL'S
INPUTS ARE SHOWN IN BLUE AND
OUTPUTS ARE SHOWN IN GREEN.
WE MADE THE SYNAPSES VARIABLE
THROUGH A CONNECTIVITY APP.
HERE IS A CELL WHERE THE APP
SHOWS THAT A LARGE NUMBER OF ITS
OUTPUTS ARE IN THIS CYAN REGION
ABLP AND THE APP ALSO SHOWS WHAT
IS THE PROBABILITY OF PARTICULAR
NERVE TRANSMITTER PHENOTYPES FOR
PARTICULAR CELL SYNAPTIC
CONNECTIONS.
YOU CAN ALSO SEE IN THE APP
LISTS OF THE UPSTREAM AND
DOWNSTREAM SYNAPTIC PARTNERS OF
THE CELL.
FOR EACH CELL WNT FLY WIRE
INTERFACE, A LIGHT BULB MENU
TAKES YOU TO ALL THE INFORMATION
ASSOCIATED WITH IT, SUCH AS A
CHANGE LOG OF WHO ARE ALL THE
PEOPLE WHO MADE PROOFREADING
EDITS, WHO MARKED THE CELL AS
COMPLETE, ALL THE PEOPLE THAT
PUT IN ANY IDENTIFYING
INFORMATION AND THINGS LIKE
THAT.
HERE IS ONE EXAMPLE OF HOW MUCH
ONE USER PROJECT CAN ACCOMPLISH
IN FLY WIRE.
IN A LAB AT CAMBRIDGE, MARKED
ALL THE NEURONS TRAVELING
THROUGH THE NECK BETWEEN THE
FLY'S BRAIN AND VENTRAL NERVE
CORD, AND TO MARK BOTH
DESCENDING NEURONS COMING DOWN
FROM THE BRAIN AND ASCENDING
NEURONS GOING UP TO THE BRAIN
AND THEN DID A DETAILED STUDY OF
DESCENDING NEURONS WHERE SHE
IDENTIFIED PRESUMABLY ALL OF
THEM BECAUSE OF THIS DETAILED
NECK TRANSECTION, MORE THAN
1300, WHICH WAS MORE THAN
PREVIOUS ESTIMATES, AND NOW WITH
THE CARD LAB HAS BEEN ABLE TO
MATCH MANY OF THEM TO GAL FORD
DRIVER LINES TO BE ABLE TO
GENETICALLY MANIPULATE NEURONAL
ACTIVITY IN THE CELLS TO CARRY
THEIR ROLE IN BAIMPLE THE YOU'LL
ALSO HEAR FROM ANOTHER FLY WIRE
POWER USER STEPHANIE HAMPEL
COMING UP IN TODAY'SCISM POSE
YUMG.
PROOFREAD NG FLY WIRE IS FAST,
ALTHOUGH SOME DENSE NEURONS TAKE
A LONG TIME, MEAN BACKBONE
PROOFREADING TIME PER NEURON IS
ONLY ABOUT ELEVEN MINUTES.
HOW WOULD THAT SCAPE UP TO
PRODUCING A CONCTOME OF TLT BRA?
IT DEPENDS ON HOW MANY CELLS
THERE ARE.
RECENTLY WE'VE RELEASED A
NUCLEUS DETECTOR WHICH FINDS
ABOUT 150,000 NEURONS IN THE
BRAIN, REQUIRING MORE THAN
20,000 PERSON HOURS TO PROOFREAD
BUT LUCKILY WE'VE GOT A BIG AND
GROWING COMMUNITY.
THE NUMBER OF WEEKLY USERS IS
ALWAYS GROWING, NOW FROM MORE
THAN 70 LABS, SO EDITS PER DAY
HAVE BEEN ACCELERATING, EVERY
POINT HERE SAN EDIT AND WE'VE
NOW PASSED 1.3 MILLION MAINLY
FOCUSED ON SAR IF THE CENTRAL
BRAIN AND HAVE PROOFREAD
TOGETHER 55,000 NEURONS AND WE
EXPECT IT TO START GOING EVEN
FASTER SINCE WE'VE JUST OPENED
UP FLY WIRE TO THE TOP PLAYERS
FROM I-WIRE THE SUN LABS'
PREVIOUS SCIENCE GAME.
HERE ARE RECENT BLOG POSTS FOR
THE PLAYERS.
IT'S NOT YET OPEN TO THE GENERAL
GAMING PUBLIC UNTIL WE ADD MORE
QUALITY CONTROL FEATURES, BUT
THE CURRENT PLIERS WHO HAVE
MOVED OVER FROM I-WIRE HAVE
YEARS OF EXPERIENCE IN EM.
TO HELP ENGAGE THEM WE'VE ALSO
ADDED FEATURES LIKE USER CHATS
AND PARTNERING PLAYERS WITH
SCIENTISTS TO UNDERSTAND
BIOLOGY.
SO FAR, EALT EXPERT PLAYERS
ONBOARDED WHO HAVE COMPLETED
MORE THAN 700 CELLS IN THEIR
FIRST FEW WEEKS.
FOR SCIENTISTS WE'RE HAPPY TO
ANNOUNCE TODAY THAT WE'LL ALSO
BE ACCEPTING APPLICATIONS FOR A
PROOFREADING SERVICE SUPPORTED
BY A NEW BRAIN INITIATIVE
GRANTED FOR DISSEMINATION.
WE PARTICULARLY ENCOURAGE FLY
LABS WORKING IN THE OPTIC LOBE
TO APPLY.
THE SERVICE IS CURRENTLY FREE
BUT WE'LL ASSESS THE COMMUNITY'S
INTEREST IN ADDITIONAL FEE-BASED
SERVICES.
FOR PLANNING ON TWO MILESTONES
FOR A WHOLE BRAIN CONNECTOME,
FIRST BEING CENTRAL BRAIN WHICH
CONTAINS ABOUT 50,000 NEURONS
AND IS ON TRACK TO BE ROW LEASED
LATER THIS YEAR AND SECOND BEING
TO ADD IN THE OPTIC LOBES, ABOUT
60,000 ADDITIONAL NEURONS.
TO SUM UP, FLY WIRE IS A
RESOURCE FOR NEURON CONNECT
ACTIVITY, MORPHOLOGY AND A
GROWING NUMBER OF CELL TYPE
IDENTIFICATIONS.
IT DISIBTS PROOFREADING OF A
WHOLE BRAIN TO THE COMMUNITY,
IT'S A LIVING DATASET VARIABLE
WHILE PROOFREADING DISPNS IT'S
APPROACHING A PROOFREAD
RECONSTRUCTION OF CENTRAL BRAIN
WITH THE WHOLE BRAIN TO FOLLOW.
THANKS FOR LISTENING.
I WOULD LIKE TO ACKNOWLEDGE OUR
AMAZING COMMUNITY IS ALL THE
PEOPLE AT PRINCETON AND AROUND
THE WORLD WHO CONTRIBUTE.
CALLING OUT SPECIAL ATTENTION TO
SVEN, JEFFERS LAB AND BRAIN
INITIATIVE.
THANKS, VERY MUCH.
>> HELLO, I'M WEI-CHUNG LEE,
BOSTON CHILDREN'S HOSPITAL AND
HARVARD MEDICAL SCHOOL.
I WOULD LIKE TO QUICKLY THANK
THE ORGANIZERS FOR THE
OPPORTUNITY TO SHARE SOME OF THE
RECENT WORK IN MY LAB, IN THIS
CASE SPEARHEADED BY A VERY
TALENTED POST-DOC, AARON KUAN.
SO FIRST I WOULD LIKE YOU TO
CONSIDER, SAY, A BIKE RIDE FROM
MY LAB AT HARVARD MEDICAL SCHOOL
TO MIT AND I KNOW THAT AS I'M
TRAVELING DOWN HUNGTD TON AVENUE
AFTER I PASS BY THE MUSEUM OF
FINE ARTS AT NORTHEASTERN
UNIVERSITY AND QDOBA I KNOW THAT
I NEED TO TAKE A LEFT TURN ONTO
MASS CHUS ET CETERA AVENUE, WHAT
WE'RE UNDERSTANDING --
MASSACHUSETTS AVENUE AND WHAT
WE'RE INTERESTING IN SNGD HOW
DOES THE BRAIN TRANSFORM SENSORY
INFORMATION INTO SERIES OF
BEHAVIORAL CHOICES SUCH AS
TURNING OR NAVIGATING.
WE TEAMED UP TOES CONSTITUTED
CAN I DECISION-MAKING PATHS,
NAVIGATIONAL.
IN THIS CASE WHAT YOU SEE IS A
MOUSE THAT'S BEEN TRAINED TO
NAVIGATE THROUGH A TEAM MAZE AND
VIRTUAL REALITY SO AT THE
BEGINNING OF THE -- ARE -- THE
TEAM MAZE, THERE IS A VISUAL CUE
THAT TELLS THE ANIMAL
SO IN THIS CASE, THIS IS
ELECTRON MIGHT BE SCOPE PI
DATASET, SPANS SIX CORTICAL
LAYERS FROM PA TO WHITE MATTER,
WE'VE DONE THIS USING A TAPE
BASED USING THREK TRANSITION
MICROSCOPY DEVELOPED, SO WE
FOCUS ON TRANSMISSION ELECTRON
MICROSCOPY BECAUSE # IT GIVES US
HIGHER SPATIAL WITH -- SIGNAL TO
NOISE.
WIDE FIELD IMAGING IS MODALITY
AND ULTIMATELY LESS EXPENSIVE IN
TERMS OF INSTRUMENTATION COSTS
COMPARED TO COMPETING SEM
METHODS CURRENTLY.
WHAT THE LAB THEN DID WAS TO
CORRESPOND THESE WITH THE
CALCIUM IMAGING WITH THOSE IN
YEN, SO THE WAY WE DO THIS IS TO
ESSENTIALLY USE THE VASCULATURE
AS A ROAD MAP USING THE SURFACE
BLOOD VESSELS ON THE SURFACE OF
THE BRAIN AND ULTIMATELY GOING
TO FINER AND FINER BLOOD VESSELS
UNTIL WE HAVE MATCHING ONE FOR
ONE CELLS AND SUB BODIES BETWEEN
THE IN VIVO IMAGING AND
MICROSCOPESCOPY -- OR IN VIVO
IMAGING ELECTRON MIEPSCOPY
DATASET.
AND THEN SUBSEQUENTLY WHAT WE
CAN DO IS RECONSTRUCT THESE
CELLS IN THE EM AND ASK WHO
THEIR PRE AND POST SYNAPTIC
PARTNERS ARE.
SO WHAT I'M SHOWING YOU HERE,
HOPEFULLY YOU CAN SEE IT EVEN
DESPITE THE BGHTD BACKGROUND --
DESPITE THE BACKGROUND S A MOVIE
OF SOME OF THE CELL WE
RECONSTRUCTED IN EM, FOR CLARITY
I'VE OMITTED SOME OF THE
COLLATERAL AXONS IN THIS CASE,
THEY'RE RANDOMLY COLORED BUT
THEIR LUMINESCENCE IS BASED ON
THE ACTIVITY OF THOSE CELLS
MEASURED IN VIVO.
WHAT YOU SEE ON THE BOTTOM
LEFT-HAND CORNER IS WHAT THE
MOUSE IS ACTUALLY SEEING IN
VIRTUAL REALITY AS IT WAS
NAVIGATING THROUGH THIS TEAM
MAZE.
SO WHAT WE HAVE HERE IS THAT
WE'RE BRINGING TOGETHER
BASICALLY BEHAVIOR FUNCTIONAL
IMAGING AND ULTIMATELY SYNAPTIC
CONNECTIVITY BRIDGING THESE
THREE LEVELS TO TRY TO
UNDERSTAND HOW NEURAL CIRCUITS
UNDERLIE FUNCTION AND BEHAVIOR.
SO WHAT DID WE FIND?
FIRST THING WE DID WAS TO LOOK
AT INHIBITORY ASSOCIATION AREA
AND FOUND ESSENTIALLY AS CHOICE
SELECTIVE AS EXCITATORY ONES AND
THIS IS IN CONTRAST TO SOME
PRIMARY SENSORY AREAS WHERE
INTERNEURONS APPEAR TO BE MORE
BROADLY.
NOW, THIS CHOICE SELECTIVITY
SUGGESTS THEY MAY BE SOMEHOW
IMPORTANT IN COMPUTATIONS THAT
THIS CIRCUIT IS PERFORMING.
THIS IS ALSO CONSISTENT WITH
RECENT DATA FROM ANN CHURCHLIN'S
LAB WHERE THEY ALSO FIND
INHIBITORY NEURONS THAT APPEAR
TO BE CHOICE SELECTIVE.
THIS IS HOWEVER IN CONTRAST TO
KIND OF MOST NETWORK MODELS THAT
IF I DRAW YOUR ATTENTION ON THE
LEFT DIAGRAM SUGGEST THAT MOST
INHIBITION IS KIND OF THIS BROAD
NONSELECTIVE INHIBITION.
SO THEN THE QUESTION BECOMES
LIKE HOW IS THIS INHIBITION THE
SELECTIVE INHIBITION, HOW IS
THAT WIRED INTO THE CIRCUIT AND
WHAT KIND OF COMPUTATIONS COULD
IT UNDERLIE?
FOR EXAMPLE, IN THE MIDDLE
DIAGRAM YOU HAVE KIND OF
INHIBITORY SELECTIVE CELLS THAT
MAY BE INHIBITING TOES
EXCITATORY CELLS FROM WHICH
THEY'RE GETTING THEIR
EXCITATION, WHEREAS THE
RIGHT-HAND SIDE, YOU SAY ANOTHER
EXAMPLE OF CONNECTIVITY WHERE
INHIBITION IS ESSENTIAL
CROSS-INHIBITION WHERE
EXCITATORY CELLS ARE INHIBITED
BY INHIBITORY CELLS THAT ARE
SELECTED FOR THE OPPOSING
CHOICE.
ULTIMATELY THIS IS WHAT I'M
GOING TO SHOW YOU WE FIND IN OUR
DATA.
SO FIRST LET'S LOOK AT THE
FUNCTIONALLY SELECTIVE
CONNECTIVITY BETWEEN EXCITATORY
TO INHIBITORY CELLS.
SO IF YOU HAVE EXCITATORY CELLS
THE TOP, ONE POSSIBILITY IS THAT
EXCITATORY CELLS MAKE MORE
CONNECTIONS THAN INHIBITORY
CELLS OF SIMILAR ARE SELECTIVITY
WHEREAS YOU COULD HAVE
EXCITATORY CELLS MAKING MORE
CONNECTIONS ONTO INHIBITORY
CELLS OF THE OPPOSITE
SELECTIVITY.
SO WHAT WE FIND IS THAT IF WE
LOOK AT ANTI-SELECTIVE VERSUS
CO-SIMILAR, EXCITATORY TO
INHIBITORY CONNECTIONS, THE
EXCITATORY TO INHIBITORY
CONNECTIVITY IS SUCH THAT IT'S
BIASED TOWARDS CONNECTIONS FROM
EXCITATORY CELLS ONTO CO-TUNED
OR SIMILARLY SELECTED INHIBITORY
CELLS AND WE CAN PLOT THIS IN A
CONTINUOUS MANNER AND WHAT WE
SEE IS THIS POSITIVE CORRELATION
BETWEEN THE PROCEED AND POST
CELL SIMILARITY AND THE NUMBER
OF DENSITY OF CONNECTIONS THAT
THEY RECEIVE.
SO WILL ULTIMATELY WE SEE THAT E
TO I CONNECTIVITY IS
CO-SELECTIVE F WE LOOK AT THE
INHIBITORY TO EXCITATORY
CONNECTIONS, WE ACTUALLY SEE THE
OPPOSITE.
P SO WHAT WE SEE IS THAT
INHIBITORY CELLS ACTUALLY
PROVIDE MORE SYNAPSIS ONTO
EXCITATORY CELLS OF THE OPPOSING
SELECTIVITY.
AND THEREFORE WE SEE THIS KIND
OF NEGATIVE CORRELATION WHEN YOU
LOOK AT THE RELATIONSHIP BETWEEN
SYNAPSE NUMBER OR SYNAPSE
DENSITY TO SIMILARITY OF
SELECTIVITY.
SO WHAT WE SEE IS THAT
INHIBITORY TO EXCITATORY
ANTI-SELECTIVE.
SO WHAT WE FIND IS THIS
INHIBITORY, THIS OPPONENT
INHIBITORY ACTIVITY MOTIF WHERE
TO THE FIRST ORDER INHIBITOR
INHIBITION IS SUPPRESSING THE
OPPOSING CHOICE IN TERMS OF
EXPRESSING EXCITATORY CELLS WITH
THE OPPOSING CHOICE BUT YOU HAVE
THIS HIGHER ORDER OF ACTION
WHERE IF YOU FOR EXAMPLE HAVE A
LEFT SELECTIVE TURN SIGNAL,
BECAUSE THERE IS INCREASED
INHIBITION ONTO THE LEFT
SELECTIVE INHIBITORY CELLS THAT
ARE SUPPRESSING THE RIGHT
SELECTIVE INHIBITORY CELLS OR
EXCITATORY CELLS, THAT CAUSES
LESS INHIBITION BACK ONTO THE
LEFT SELECTED EXCITATORY CELLS.
SO WE BELIEVE THAT THIS HELPS TO
SEPARATE KIND OF THE POPULATION
DYNAMICS IN THE CIRCUIT AND BIAS
THE CHOICE OF THE ANIMAL TOWARDS
ONE SIDE OR THE OTHER AS THEY
KIND OF NAVIGATE THROUGH THIS.
SO IN THE INTEREST OF TIME, I'M
GOING TO JUST QUICKLY TALK ABOUT
CIRCUIT MODELING, TOGETHER WITH
STEFANO AND J CU LIO WHAT THEY
DID WAS DEVELOP AND TRAIN AN
ARTIFICIAL RECURRENT NEURAL
NETWORK WITH THE SAME NUMBER OF
CELLS THAT WE RECORDED IN VIVO
BUT THEY WERE KIND OF THEIR
CONNECTIVITY RATES WERE
INITIALLY RANDOMIZED BUT THEN
TRAINED THE WHOLE ENTIRE
ARTIFICIAL NEURAL NETWORK WAS
TRAIBLED ON RECA PILT LATE --
TRAINED TO RECAPITULATE THE
ACTIVITY RATE IN VIVO AND WE SEE
WE RECAPITULATE THIS COMPONENT
INHIBITORY MOW MOTIF WHERE YOU
HAVE EXCITATORY TO INHIBITORY
CONNECTIVITY THAT IS
CO-SELECTIVE AND INHIBITORY TO
EXCITATORY CONNECTIVITY THAT IS
ANTI-SELECTIVE SO WE THINK THAT
THIS IS KIND OF A GENERAL
PRINCIPLE OF NEURAL NETWORKS
THAT ARE INVOLVED IN THINGS LIKE
OR TASKS LIKE DECISION-MAKING.
SO IN THE INTEREST OF TIME, I'M
GOING TO END HERE BUT BASICALLY
THIS IS TELLING US THAT OR IT
SUGGESTS THAT DIFFERENT CORTICAL
AREAS ARE ESSENTIALLY MAY HAVE
DIFFERENT CONNECTIVITY ROLES
THAT IT COULD PERFORM AND
UNDERLIE DIFFERENT FUNCTIONS
WHERE YOU HAVE SELECTIVE
CONNECTIVITY OF INHIBITORY CELLS
IN THE PARIETAL CORTEX COMPARED
TO BROADLY ATTUNED MORE RANDOM
CONNECTIVITY IN PRIMARY SENSORY
AREAS WHICH MAY BE BETTER SUITED
FOR GAME CONTROL AND FINALLY I
WOULD LIKE TO THANK THE PEOPLE
IN MY LAB THAT PERFORMED AND
HELPED WITH THIS STUDY AS WELL
AS THE FUNDING AND SUPPORT THAT
WE'VE RECEIVED.
SO THANK YOU.
>> THE ABILITY TO PRODUCE
COMPLEX BEHAVIORS BY ASSEMBLING
SEQUENCES OF DIFFERENT MOVEMENTS
IS ESSENTIAL FOR PURPOSEFUL
BEHAVIOR AND SURVIVAL.
OUR LAB IS INTERESTED IN HOW
MOVEMENT SEQUENCES ARE GENERATED
BY THE NERVOUS SYSTEM.
WE STUDIED GROWING BEHAVIOR IN
FRUIT FLIES AS MODEL TO
UNDERSTAND HOW SUCH SEQUENCES
ARE PRODUCED, AND YOU MIGHT
WONDER WHY GROWING BEHAVIOR,
BECAUSE THE CONSIST OF THE
STEREOTYPE SEQUENCE OF MOVE
MGHTS THAT RULE THE DIFFERENT
BODY TYPES.
SO WE USED GROOMING AND FRUIT
FLIES OR FRUIT FLIES
SPECIFICALLY BECAUSE OF THE
TOOLS THAT ENABLE US TO IDENTIFY
AND MAP NEURAL CIRCUITS RS AND
THE SYNAPTIC RESOLUTION.
ONE WAY WE CAN EASILY ELICIT
GROWING BEHAVIOR IN THE LAB IS
BY CODING FLIES AND WHAT THIS
DOES IS CAUSES THE FLY TO MOVE
TO CLEAN ITS WHOLE BODY USING
DISTINCT GROOMING MOVEMENTS.
LET ME SHOW YOU WHAT I MEAN BY
THIS.
P IN THE VIDEO ON THE LEFT, YOU
CAN SEE A FLY A DIRTY FLY USES
FRONT LEGS TO GROOM ITS HEAD.
BUT WE FOUND THAT THE FLY CAN
ACTUALLY GROOM SPECIFIC PARTS OF
THE HEAD SUCH AS ONLY ITS EYES
OR ANTENNA OR MOUTHPARTS.
AND ON THE RIGHT YOU CAN SEE A
DIRTY FLY USING ITS HIND LEGS
USING ITS POSTERIOR BODY AND AT
THE MOMENT IT'S GROOMING ITS
ABDOMEN AND IT'S SWITCHING RIGHT
NOW TO ITS SWINGS.
SO IN OUR PREVIOUS WORK, WE
FOUND THAT FLIES GROOM IN AN
ANTERIOR TO POSTERIOR GROOMING
SEQUENCE THAT STARTS WITH THE
EYES, PROCEEDS TO THEIR ANTENNA,
MOUTHPARTS, ABDOMEN, WINGS, AND
WE ALSO PREVIOUSLY DISCOVERED
THAT THE GROOMING SEQUENCE IS
PRODUCE BID A PARALLEL MODEL OF
HIERARCHICAL SUPPRESSION WHICH
YOU CAN SEE HERE.
I'M NOT GOING TO GO INTO MUCH
DETAIL OF THIS MODEL BUT I
DOMENT TO POINT OUT A COUPLE OF
FEATURE -- BUT I DO WANT TO
POINT OUT A COUPLE OF FEATURES
THAT ARE IMPORTANT TO UNDERSTAND
FOR TODAY'S PART.
WHEN WE DUST THE PARTICULAR BODY
PART SUCH AS THE 5BD, ONLY THAT
MOVEMENT GETS ACTIVATED.
HOWEVER, WHEN FLIES ARE --
DIFFERENT MUTE OOL DIFFERENT
MUTUALLY EXCLUSIVE BODY
MOVEMENTS THAT CLEAN DIFFERENT
PARTS OF THE HEAD OR BODY
ACTIVATED IN PARALLEL BY
MECHANIC AN NOTICE SENSORY
NEURONS THAT DETECT DUST ON THAT
BODY PART.
SO THE NEED TO PERFORM ALL OF
THESE DIFFERENT MOVEMENTS MEANS
THAT THEY ARE ALL COMPETING FOR
OUTPUT, BUT ONLY ONE MOVEMENT
CAN BE PERFORMED AT A TIME.
SO THIS GETS RESULTS THROUGH
HIERARCHICAL SUPPRESSION WHEREBY
EARLIER MOVEMENTS SUPPRESSION
LATER ONES.
THIS SO-CALLED PARALLEL SMODLE
OF GENERAL INTEREST ACTUALLY
BECAUSE SIMILAR MODELINGS HAVE
BEEN USED TO DESCRIBE HOW
DIFFERENT BEHAVIOR SEQUENCE RS
PRODUCE AND HAD THIS MODEL
PROVIDES A GREAT CONCEPTUAL
FRAMEWORK FOR US FOR MAPPING THE
NEURAL CIRCUITS THAT PRODUCE THE
GROOMING SEQUENCE, WHICH IS THE
CURRENT FOCUS OF OUR LAB.
IN THE WORK I'M PRESENTING TODAY
I WILL ONLY FOLK TUS ON THE
INPUT LAYER, BASICALLY THE
MECHANO-SENSORY NEURONS THOUGHT
TO DETECT THE DUST THAT
ACTIVATES EACH MOVEMENT IN
PARALLEL.
I WILL JUST FOCUS ON THE FLY
HEAD AND SHOW YOU HOW WE MAP
MECHANO SENSORY LOCATIONS FROM
DIFFERENT LOCATIONS ON THE HEAD
THROUGH DIFFERENT NEURONS INTO
THE BRAIN.
IN OUR PREVIOUSLY PUBLISHED AND
UNPUBLISHED LISHED WORK, WE HAVE
SHOWN THAT THE RELATION --
PUBLISHED AND UNPUBLISHED WORK
WE HAVE SHOWN THAT THE RELATION
OF CYTOMEGALY RESPONSES PROVIDE
THE GROOMING.
INROAD INTO THE GROOMING
NETWORK.
WHAT WE HAVE DONE IS TO
IDENTIFY, CLASSIFY AND QUANTIFY
ALL MECHANO SENSORY ON THE FLY
HEAD.
HERE WE USED A DRIVER LINE THAT
LABELS ALMOST ALL MECHANO
SENSORY NORMS, WHICH I WILL
REFER TO FOR THE REST OF THE
TALK.
WE IMAGED HEADS USING MICROSCOPE
AND WHAT WE OBSERVED WAS THAT
EACH BRISTLE IS IN-- IS
INNERVATED BY A SINGLE NEURON.
WE THEN RECONSTRUCTED THOSE ENDS
AND THEIR PROJECTIONS FROM THEIR
HEAD LOCATIONS IT THE BRAIN --
THEIR HEAD THROAKSZ THE BRAIN.
THIS REVEALED DIFFERENT PARTS OF
THE HEAD PROJECT TO THE BRAIN
THROUGH FOUR DIFFERENT NERVES.
HERE YOU CAN SEE THE
RECONSTRUCTED NERVES, NEURONS
AND THEIR NERVES IN DIFFERENT
COLORS, THE ANTENNA, ANTERIOR
HEAD, IN GREEN THE OCCIPITAL
NERVE, IN RED THE EYE NERVE,
BOTH ON THE POSTERIOR PART OF
THE HEAD AND PROJECT FROM
PROBOSCUS AND ALL THESE GO INTO
A REGION OF THE BRAIN SAZ.
IF YOU LOOK AT THE PROJECTIONS
TO GET THERE IN THIS DRIVER LINE
THAT TARGETS ALMOST ALL BNM'S,
IT IS IMPOSSIBLE TO DISTINGUISH
MOTOR NEURONS OR EVEN THE
DIFFERENT NERVES.
SO TO BE ABLE TO INDEPENDENTLY
LABEL AND VISUALIZE THE
DIFFERENT NERVES WE BUILD
DIFFERENT TRANSGENIC DRIVER LINE
AND HERE YOU CAN SEE THESE FOUR
LINES THAT LABEL NEURONS FROM
EITHER ONE NERVE IN THESE TWO
LINES INDEPENDENTLY OR TWO
NERVES IN THESE TWO LINES
INDEPENDENTLY, WHICH MAKES IT
MUCH EASIER TO VISUALIZE THE
NERVE PROJECTIONS INTO THE
BRAIN.
HOWEVER, WE NEXT WANTED TO
DETERMINE THE MORPHOLOGY OF
INDIVIDUAL BEINGS THAT INNERVATE
DISTINCT BRISTLES.
FOR THIS WE PERFORMED DYE FIELDS
THAT INNERVATE THE LARGEST
BRISTLES THE ON HEAD.
SO WHAT WE DID IS WE PLUCKED THE
POLICE HE WILL AND ADDED DYE
INTO THE EXPOSED SOCKET THAT
CONTAINS THE DENDRITE OF THE
ASSOCIATED BNM.
THE DYE THEN DIFFUSED INTO THE
NEURONS AND THE RESULTED YOU CAN
APPRECIATE HERE ON THE RIGHT
LABELED BNM'S.
FOR BRISTLES WE COULDN'T DYE, WE
USED VERY SMALL BRISTLES ON THE
BACK.
HEAD, WE USED STOIK --
STOICHIOASTICALLY GUIDELINES IN
THE DRIVER LINES.
SAME POPULATIONS HAVE VERY
SIMILAR MORPHOLOGY AS WE CAN SEE
HERE IN THESE ANTENNAE ENDS THAT
INNERVATE FOUR DISTINCT
BRISTLELESS ON THE ANTENNAE.
INTERESTINGLY, WHEN WE COMPARED
PROJECTIONS OF BNM ENDS THAT
INNERVATE NEIGHBORING
POPULATIONS, WE FOUND THEY ALSO
HAVE SIMILAR MORPHOLOGY.
SO NOW WE KNOW A LOT ABOUT THE
MORPHOLOGY OF INDIVIDUAL NEURONS
MPLETS HOWEVER, TO DEFINE THEIR
ORGANIZATION IN THE BRAIN, WE
NEEDED TO BE ABLE TO LOOK AT ALL
BNM'S TOGETHER WHILE KNOWING THE
IDENTITY OF THE BRISTLE THEY
INNERVATE.
SO FOR THIS, WE LOOKED IN A
SECTION ELECTRON MICROSCOPE
VOLUME OF AN ENTIRE FRUIT FLY
BRAIN FOR EACH OF THE NERVES TO
RECONSTRUCT ALL MECHANO SENSORY
NEURONS OF THE HEAD.
FOR THIS WE USED THE FLY WIRE AI
PLATFORM TO SEE EACH ENTIRE
NERVE AND RECONSTRUCT ALL
NEURONS, AND I'M SHOWING YOU ONE
EXAMPLE HERE, WHICH IS THE EYE
AND THE CAN -- NERVES MERGED
TOGETHER WEVMENT SEE THE ENTIRE
DISMERVES CONSTRUCTED ALL
NEURONS AND FROM THIS WE
IDENTIFIED 701 BNM'S OF THE
RECONSTRUCTED NEURONS.
SO WHAT WE FOUND IS THAT THE
BNM'S ARE SEW MATT TOPICALLY
ORGANIZED AS THEY'RE DISTINCT
PROJECTIONS FROM DIFFERENT
NERVES CORRESPOND TO DIFFERENT
BRISTLE LOCATIONS ON THE HEAD.
HERE YOU CAN SEE A FLY BRAIN AND
DIFFERENT ORIENTATIONS, THE
LATERAL, DORSAL AND ANTERIOR
VIEW, AND THE NERVE PROJECTIONS
MAGNIFIED AT THE BOTTOM HERE.
SO FOR EXAMPLE, BNM'S THAT
INNERVATE BRISTLES ON THE
PROBOSCIS PROJECT TO A DIFFERENT
AREA IN THE SEC THAN BNM'S THAT
INNERVAT THE EYES HERE.
I DON'T HAVE MUCH TIME TO SHOW
YOU THIS TODAY, BUT WE INCREASED
THE RESOLUTION OF THE SEW MATT
OH TOPIC MAP -- SOMATOTOPIC MAP
A LOT, ALMOST ALL THE BRISTLES
ON THE HEAD TO RECONSTRUCTED
NEURONS.
LET ME SUMMARIZE WHAT I JUSTBOU.
WE IDENTIFIED NEARLY ALL BNM'S
ON THE DROSOPHILA HEAD THAT
ELICITED GROOMING OF SPECIFIC
HEAD PARTS, WE RECONSTRUCTED BNM
PROJECTIONS FROM DIFFERENT
LOCATIONS ON THE HEAD TO THEIR
DISTINCT AREAS IN THE BRAIN
WHICH PROVIDES THE FIRST
SYNAPTIC RESOLUTION'S METATOPIC
MAP OF THE HEAD AND NEUROSENSORY
PATHWAYS WHICH IS HEAD FORMING,
GROUNDWORK TO ELUCIDATE PARALLEL
ARCHITECTURE THAT UNDERLIES EACH
OF THE HEAD GROOMING MOVEMENTS.
THIS IS ALL A MAJOR CONTRIBUTION
TO FLY WIRE TO INCORPORATE INTO
THEIR FLYI BRAINULD LIKE TO THL
THE PEOPLE THAT HAVE BEEN
INVOLVED IN THE PROJECT,
ESPECIALLY ANDY, CATARINA AND
ANDIVA WHO WERE THE MAIN
CONTRIBUTORS TORS, NIH BRAIN
INITIATIVE AND NIH CORPORATE WHO
ARE FUNDING THIS PROJECT, AND I
WOULD LIKE TO THANK YOU FOR
INVITING ME TO PRESENTED TODAY
AND FOR YOUR ATTENTION.
THANK YOU.
>> GOOD OOIVE, EVERYONE -- GOOD
AFTERNOON, EVERYONE, MY NAME IS
MICHAEL REISER, GROUP LEADER, MY
WORK IS ON -- FLIES.
TODAY I'LL TELL YOU ABOUT MY
LAB'S EXPLORATION FOLLOWING
VISUAL PATHWAYS OUTSIDE OF THE
VISUAL SYSTEM WHICH IS A
SLIGHTLY UNUSUAL THING TO DO AND
I'LL TELL YOU ABOUT IT AS WE
MOVE ON AND THE WAY WE'VE
APPROACHED THIS PROBLEM IS BY
COMBINING FUNCTIONAL
MEASUREMENTS WITH CONNECTOMIC
ANALYSIS.
AND SO AS YOU MAY KNOW, VISION
IS VERY PROMINENT IN THE BRAIN
OF SIGHTED ANIMALS AND WHEN WE
SEE THAT BY LOOKING FOR EXAMPLE
AT THE AMOUNT OF CORTICAL REAL
ESTATE DEDICATED TO VISION IN MY
FAVORITE ORGANISM IT THE FLY,
CROSS-SECTION OF THE FLY BRAIN
AND LOOK AT AGAIN REAL ESTATE,
WE FIND THAT HALF THE BRAIN IS
DEDICATED TO VISION AND THAT'S
WHETHER YOU THINK ABOUT VOLUME
OR THE NUMBER OF NEURONS.
THINKING ABOUT THIS, ONE OF THE
REASONS WHY VISION TAKES UP SO
MUCH SPACE IS BECAUSE OF RETINA,
HERE IN THE FLY BRAIN WE CAN SEE
THAT IF WE FOLLOW THE VISUAL
INFORMATION FROM A PARTICULAR
POINT IN SPACE AND WE FOLLOW IT
THROUGH COLUMNAR NEURONS THROUGH
DIFFERENT NEURAL FIELDS IN THE
VISUAL SYSTEM WHAT WE HAVE IS WE
HAVE SETS OF NEURONS WHICH ARE
DEDICATED TO IMAGING INFORMATION
FROM A PARTICULAR REGION AND
SPACE AND THEY MAINTAIN THE
SPATIAL RELATIONSHIP BETWEEN
DIFFERENT PARTS OF THE VISUAL
FIELD WHILE FEATURES ARE BEING
EXTRACTED.
SO THIS ORGANIZATION HAS BEEN
INCREDIBLY USEFUL, FOR EXAMPLE,
IN STUDYING THE ORIGIN OF
DIRECTIONAL SELECTIVITY AND THE
ORGANIZATION OF MOTION VISION.
SO WHAT WE'VE BEEN WONDERING IS
WHAT HAPPENS WHEN YOU LEAVE THE
VISUAL SYSTEM?
SO THAT ORGANIZATION OF VISUAL
RETINA -- HAS BEEN VERY HELPFUL
IN THINKING ABOUT FEATURE
DETECTION, BUT WHAT HAPPENS WHEN
YOU NO LONGER HAVE THIS WELL
ORGANIZED STRUCTURE.
THAT'S WHAT WE'VE BEEN LOOKING
AT ASK DOING THIS RECENT BILL
FOCUSING A SET OF NEURONS CALLED
LC NEURONS, NEURONS WHICH
RECEIVE THEIR INPUTS HERE IN THE
LOBULA WHERE THEY HAVE DENDRITES
IN THIS CELL TYPE LC6 CASE WHICH
CORE TO UNDERSTAND TO 4 PERCENT
OF THE FIELD OF THE FLY AND
DENDRITES ARE ORGANIZED AND
RECEIVE INPUTS FROM RETINA
TOPIC, CONNECT TO GLOMERULUS
WHERE A LOT OF THAT ORGANIZATION
IS NO LONGER EVIDENT AS YOU CAN
SEE HERE.
THERE'S BEEN THE PROPOSAL THIS
REPRESENTS SOME ABSTRACTION OF
VISUAL INFORMATION BUT THAT'S
THE OPEN QUESTION.
THAT'S WHAT WE'VE BEEN THINKING
ABOUT IS HOW DOES VISION GET
SORT OF REFORMATTED AS YOU CROSS
THIS LAYER OF THE BRAIN WHERE
YOU GO FROM VERY WELL ORGANIZED
VISUAL STRUCTURES TO NONVISUAL
OR NOT OBVIOUSLY VISUAL
ORGANIZATION.
SO WE'VE APPROACHED THIS PROBLEM
IN THE WAY ME AND A LOT OF MY
COG LEAGUES HAVE A-- COLLEAGUES
HAVE APPROACHED MANY PROBLEMS IN
THE FLY WHICH IS FIRST BY
STARTING WITH A SYSTEMATIC SET
OF GENETIC DRIVER LINES WHICH
ALLOW US TO TARG THE NEURONS ONE
CELL TYPE AT A TIEMENT WHICH
ALLOWS US TO DO THESE SORT OF
HIGH-QUALITY ANATOMIC
DESCRIPTIVE STUDIES AND THEN WE
ALSO HAVE TOOLS WE CAN USE FOR
FUNCTIONAL STUDIES.
SO WE FOCUS QUITE A BIT ON LC6
WHICH WAS THE FIRST NEURON I
JUST SHOWED AND THE ONE WE USE
TO THINK ABOUT THE ORGANIZATION
WITHIN A GLOMERULUS, LC6 IS THE
ONE HERE IN BLUE.
AS YOU CAN SEE THERE ARE MANY
GLOMER LIE AND THEY PROJECT
IN -- GLOMERULI AND THEY PROJECT
IN PARALLEL INTO THE BRAIN.
FLY.
I WANT TO MENTION THAT PUBLISHED
A FEW YEARS AGO WE HAD A STUDY
WHERE WE LOOKED AT THE
ORGANIZATION WITHIN A
GLOMERULUS, FOLLOWED NEURONS
INTO THE GLOMERULUS, FOUND THEIR
TARGETS AND THOUGHT ABOUT HOW
WITHIN THE GLOMERULUS THE
READOUT, WHAT KIND OF
INFORMATION IS IN THAT READOUT
FROM A GLOMERULUS AND THEN THE
STUDY I'LL TELL YOU ABOUT TODAY
IS ONE THAT WAS JUST PUBLISHED A
MONTH AGO WHERE THIS WAS A STUDY
LED BY NATHAN WHO IS A JOINT
POST-DOC AND WE WERE LOOKING AT
THE READOUT ACROSS GLOMERULI AND
MIGHT 2E8 YOU HOW THEY ARE ORG
NIERLTZED AND WHY THEY ARE WHERE
THEY ARE.
IN BOTH STARTED WITH FUNCTIONAL
IN THE LAB AND VERY QUICKLY
REALIZED THAT THESE STUDIES
WOULD BE POWERFULLY INFORMED BY
CONNECTOMIC DATASET.
IN THE FIRST INDICATE WE
MANUALLY RECONSTRUCTED LC
'6NEURONS AND THEIR TARGETS INTO
THE GLOMERULUS AND CENTRAL BRAIN
AND SECOND STUDY WE WERE ABLE TO
TAKE ADVANTAGE OF THE NOW VERY
FAMOUS HEMIBRAIN CONNECTOME TO
LOOK AT THE CONNECTIVITY OUT OF
THESE GLOMER DAIRN GLOMERULI
WITHIN THE CENTRAL BRAIN.
S EVEN THOUGH QUITE A FEW THINGS
HAVE BEEN SAID ABOUT THESE SINCE
WE PUBLISHED THAT PAPER SIX
MONTHS AGO THERE'S A LOT WE
DON'T KNOW ABOUT THEIR EN
CAIDING SO NATHAN TOOK ON THIS
FLOB A SOPHISTICATED WAY BY
DOING VOLUME MELT TRICK IMAGING,
TWO PHOTON VOLUMETRIC IMAGING,
NOT IN GLOMERULUS OR VISUAL
SYSTEM BUT IN THE REGION BETWEEN
THEM WHERE THE NEURONS CAN BE
INDIVIDUALLY SEGMENTED ALLOWING
YOU TO DO A POWERFUL
EXPERIMENTAL APPROACH WHERE YOU
CAN MAP RECEPTIVE FIELDS OF
INDIVIDUAL NEURONS, DISTINGUISH
THEM BASED ON THE
THREE-DIMENSIONAL ROI'S FOR THE
NEURONS AND THEN YOU CAN DELIVER
VISUAL STIMULI CENTERED ON THE
RECEPTIVE FIELD CENTER WHICH HAD
BEEN MAPPED OUT FOR EACH NEURON
AND THAT GIVES YOU ACCESS TO A
LEVEL OF DESCRIPTION OF THE
FUNCTIONAL CELL ACTIVITY OF
KNEES NEURONS WHICH HAD NOT BEEN
PREVIOUSLY POSSIBLE.
SO A SIMPLE THING YOU WOULD
MEASURE DOING THIS IS JUST ASK
HOW LARGE ARE THE RECEPTORS OF
THESE NEURONS AND YOU CAN DO
THAT HERE AND SEE THAT WHEN WE
COMPARE THIS FUNCTIONAL
MEASUREMENT TO THE ANATOMICAL
MEASUREMENT, WE FIND EXCELLENT
CONCORD ANSWER.
SNRIET IN GENERAL, NEURONS WITH
LARGER DENDRITES HAVE LARGER
RECEPTIVE FIELDS, BUT THAT'S THE
KIND OF -- THAT'S NOT REALLY THE
OPTIMIZED THING FOR THESE
NEURONS, THIS IS JUST THE
MAPPING STIMULUS BUT WHAT NATHAN
DID IS A SYSTEMATIC WAY DELIVER
A WHOLE PANEL OF STIMULI TO
THESE NEURONS WHICH ALLOWED HIM
TO DESCRIBE THE RESPONSE THROUGH
A WHOLE RANGE OF STIMULI, HERE
IS AN EXAMPLE FOR ONE CELL TYPE
BUT THEN HERE IS THE SET OF THE
TEN LC'S THAT NATHAN LOOKED AT
CAREFULLY AND WHAT WE FIND AGAIN
THESE ARE STIMULI DELIVERED TO A
CENTER OF THE RECEPTOR FIELD
MAPPED FOR EACH INDIVIDUAL
NEURON WHAT WE FIND IS THAT AT
THE HIGH LEVEL YOU CAN CUT THESE
NEURONS INTO TWO GROUPS, A GROUP
THAT PRIMARILY RESPONDS TO SMALL
OBJECT MOTION AND A GROUP THAT
PRIMARILY RESPONDS TO VISUAL
LUMEN SO EXPANDING THIS STIMULI
AND THEN WITHIN THE GROUPS
THERE'S A LOT OF DETAILS WHICH
I'LL POINTED YOU TO THE PAPER TO
LOOK INTO FURTHER.
BUT THAT IS JUST KEEP THAT IN
MIND AT THE FIRST LEVEL OF THIS
SMALL OBJECT.
SO THEN WE THEN GO BACK TO THIS
QUESTION OF ASKING HOW ARE THE
GLOMERULI ORG NIEDZED AND IT'S
IMPORTANT TO POINT OUT THEY DO
FORM THIS VISUALLY CONFIGUREOUS
GROUP OF GLOMERULI IN THE BRAIN
AND WE CAN START TO IMPOSE THE
LAINLZ, THESE FUNCTIONAL LABELS
BACK ONTO THE NEURONS AND INDEED
SO THE LABELS OF THE LUMEN GROUP
AND OBJECT GROUP AND WE FIND WE
ALREADY HAVE A PRETTY GOOD CLUE
THAT LOOMING NEURONS ARE NEXT TO
LOOMING NEURONS AND ANOTHER WAY
TO SEE THAT IS BY LOOKING AT
PRINCIPAL COMPONENT ANALYSIS OF
ALL THE RESPONSES ACROSS ALL THE
MEASUREMENTS IN THAT DATASET AND
HERE EACH DOT REPRESENTS THE
RESPONSES OF A SINGLE NEURON AND
YOU CAN SEE THAT ALL THE NEURONS
CO-CLUSTER, SO ALL OF THE
INDIVIDUALS OF ONE CELL TYPE
HAVE RESPONSES THAT ARE MORE
SIMILAR TO THE OTHER NEURONS OF
THE CELL TYPE, AND WE ALSO FIND
THAT THE SPATIAL POSITION OF THE
NEURONS IN THE BRAIN LOOKS QUITE
A BIT LIKE HOW THEY'RE POSITION
IN THIS PCA SPACE SO FOR EXAMPLE
IF YOU'RE LC15 AND LC25 ARE
CLOSE AND AGAIN LPLC2 AND 4 ARE
ALSO CLOSE NOW AND RARELY ARE IN
MY HANDS ARE A PCA ANALYSIS,
PRINCIPAL EXEENLT CIS, HERE
INDEED IT REALLY LOOK AS IF
PRINCIPAL COMPONENT ONE IS
DESCRIBING THE NEURON'S RESPONSE
SIFT TO LUMEN WHEREAS THE SKD
PRINCIPAL COMPONENT IS LARNLLY
DESCRIBING THE NEURON'S RESPONSE
TO SIZE, WHAT ELSE THE OPTIMAL
BEST SIZE STIMULUS WHICH DRIVES
THE NEURON.
IT I HOPE I'VE CONVINCED YOU
THAT NEARBY BINEURONS HAVE MORE
SIMILAR RESPONSES, THESE
GLOMERULI ARE ORGANIZED INTO A
FEATURE MAP AND WE WANT TOP
WHETHER THE BRAIN CAN USE THAT
FEATURE MAP AND LOOK AT
DOWNSTREAM NEURON AND ASK
WHETHER THEIR READOUT SUGGESTS
THAT THEY'RE READING FROM
SPECIFICALLY NEARBY GLOMER LIE
AND WE DO THAT WITH CONNECTOME,
USING HEMIBRAIN TO DO THIS AND I
CAN'T SHOW YOU HOW REMARKABLY
STEREOTYPED THESE BRAINS ARE.
TOP IMAGES FROM LIGHT MICROSCOPY
BASED ON LABELING P BARS,
PRESYNAPTIC ACTIVE ZONES OF
NEURONS, A COMPOSITE OF TEN
DIFFERENT BRAINS, WE HAVE
ANOTHER BRAIN, ELEVENTH BRAIN
WHICH IS THE ONE THAT THE
CONNECT TOMORROW EXISTS FOR AND
WE JUST COLOR CODE IT IN THE
SAME WAY AND I THINK IF YOU
STARE AT THIS FOR A WHILE YOU'LL
SEE EVEN AT THE FINE DEGREE
THERE'S A HUGE CONCORDANCE
BETWEEN THESE TWO DATASETS, IT
ALLOWS US TO MAKE THESE LEAPS
CROSS DIFFERENT ANIMALS AND
USING THIS CELL TYPE AS THE
CURRENCY FOR THE COMPARISON THAT
WE CARRY ACROSS.
SO NOW FOR THIS QUESTION.
ARE THE NEIGHBORING GLO ME RULI
IN A VISUAL MAP USED IN SHARED
COMPUTATION?
WE USE THIS, WE CAN JUST ASK
WHETHER NEURONS THAT READ OUT
FROM PAIRS OF COLUMNAR CELLS ARE
THEY MORE LIKELY TO READ OUT
FROM GLOMERULI THAT ARE CLOSE
BY?
INDEED THEY ARE.
PAIRINGS ARE SIGNIFICANTLY
SKEWED TOWARDS NEARBY GLOMERULI
AS OPPOSED TO JUST RANDOMLY
SHUFFLE AND POST CONNECTIONS.
APPEARS I MIGHT SUSPECT, THE
PROXIMITY AND FUNCTIONAL
SIMILARITY, INDEED WE FIND
SLAIRL THESE PAIRINGS ARE SKEWED
TOWARDS INPUTS THAT ENCODE
SIMILAR VISUAL FEATURES. SO WE
FIND THAT ON AVERAGE, THE
TARGETS IN THE CENTRAL BRAIN ARE
REEKED OUT FROM NEARBY GLOMERULI
WITH SIMILAR FEATURES, THIS IS A
HIGH LEVEL RESULT AND WHAT'S
BEAUTIFUL IS THAT THEN YOU HAVE
ACCESS TO ALL THE DETAILS.
SO INDEED WE CAN LOOK AT ALL THE
PAIRS THAT EXIST IN THE BRAIN,
THERE'S A SUMMARY HERE WHICH
I'LL POINTED YOU TO THE PAPER
FOR MORE DETAILS, BUT THEN WE
CAN USE THIS TO PROPOSE NEW
EXPERIMENTS.
SO HERE ARE SPECIFIC NEURONS IN
THIS CASE A NEURON WHICH OUGHT
TO BE QUITE SENSITIVE TO VISUAL
LOOMING, SHOULD BE QUITE SENSE
TOIF TO OBJECT MOTION BY
PROPERTIES OF ITS INPUT AND HERE
IS A NEURON THAT INTEGRATES A
LOOMING SENSITIVE PATHWAY AND
ONE FROM A SMALL OBJECT PATHWAY,
HOWEVER IT INTEGRATES FROM THE
FASTER NEURONS FROM EACH
PATHWAY, INTERESTING HYPOTHESIS
THERE.
WE CAN SEE THIS CONNECTOME
ANALYSIS QUICKLY FEEDS BACK TO
THE NEXT ROUND OF EXPERIMENTS WE
WOULD WANTED TO DO.
TO QUICKLY SUMMARIZE AS I'M
RUNNING OUT OF TIME, WE'VE BEEN
LOOKING ALTHOUGH VISUAL
PROJECTIONS IN THE FLY BRAIRNGS
DOZENS OF PARALLEL PATHWAYS THAT
GO INTO THE CENTRAL BRAIN, BEING
ENCODES DISTINCT VISUAL
FEATURES.
WE WERE ABLE TO READ OUT SOME
SPATIAL INFORMATION EVEN THOUGH
IT'S COARSER THAN WHAT YOU FIND
IN THE RETINOTOPIC FIELDS AND
ACROSS GLOMERULI WE FIND THAT
MORE SIMILAR ENCODING NEURONS
HAVE GLOMERULI THAT ARE CLOWER
TO EACH OTHER AND WE REALLY FIND
THERE'S ORGANIZATION EVERYWHERE,
NOTHING IS RANDOM OF THE LAST
POINT I LIKE TO LEAVE PEOPLE
WITH IS WHAT THIS HAS TAUGHT US
IS THESE PATHWAYS FOR VISUALLY
GUIDED BEHAVIORS ARE DEEPER THAN
PEOPLE EXPECTED, EVEN IN SIMPLE
FLY BRAIN, BUT WE HAVE ALL THE
TOOLS AND MORE CONNECTOMES
ONLINE TOMG CONGRESS UP TO HELP
US MAP THESE PATHWAYS WHICH IS
CONVERTED INTO BEHAVIOR WHICH IS
A TOPIC I'M EXCITED TO TALK
ABOUT AND WHAT I DO WITH THIS
AMAZING GROUP OF PEOPLE.
THANK YOU VERY MUCH FOR YOUR
TIEMENT, I LOOK FORWARD TO
THE -- FOR YOUR TIME, I LOOK
FORWARD TO THE DISCUSSION.
>> GOOD AFTERNOON.
SYNAPSES ARE THE HALLMARK OF THE
BRAIN AND 95 PERCENT OF
MAMMALIAN BRAIN SYNAPSES ARE
EXCITATORY AND IT'S RECENTLY
BECOME APPARENT THAT THERE'S A
VAST DIVERSITY OF EXCITATORY
SYNAPSE TYPE THAT DIFFER IN
THEIR MOLECULAR COMPOSITION.
THIS SYNAPSE DIVERSITY IS
DESCRIBED BY THE WORD SYNAPTOME
AND IMPORTANTLY THERE'S A
MOLECULAR LOGIC OF THE SYNAPSE
DIVERSITY AS SHOWN HERE WHERE
GENES ENCODE OBVIOUSLY RNA AND
PROTEINS, WHICH IN TURN ARE
BUILDING BLOCKS FOR BLOCKS FOR
COMPLEXES AND THOSE ARE BUILDING
BLOCKS FOR SUPER-COMPLEXES AND
IT IS THE COMPLEX AND IS SUPER
COMPLEXES THAT ARE DISTRIBUTED
INTO SYNAPSE TO SAY GIVE THEM
THEIR DIFFERENT MOLECULAR
COMPOSITIONS, THESE ARE REALLY
THE BUILDING BLOCKS FOR THE SNAP
SYNAPTOME.
THEN SPATIAL AT THE CELLULAR
LEVEL, DIFFERENT TYPES OF
SYNAPSES ARE DISTRIBUTED AND
THOSE DIFFERENT CELL TYPES THEN
POPULATE DAIFNT PARTS OF THE
BRAIN TO GIVE AN OVERALL
BRAIN-WIDE SYN APPEAR TOME BRAIN
ARCHITECTURE.
METHODS FOR EXAMINE INDIVIDUAL
SYNAPSES IN MAMMALIAN BRAIN ON
BRAIN WIDE SCALE AS SHOWN IN THE
TOP HERE INVOLVE, INVOLVE
TARGETING FLUORESCENT PROTEINS
TO ENDOGENOUS MOLECULES,
DIFFERENT TYPES OF COME PLEKS,
BY EXAM ANYONING THEIR
DISTRIBUTION SYNAPSES USING
MICROSCOPY, FOR EVERY INDIVIDUAL
WE CAN QUANTIFY A VARIETY OF
PARAMETERS EXPRESSION OF THE
PROTEIN AND SIZE AND SHAPE
PARAMETERS USING MACHINE
LEARNING DATA FROM BILLIONS OF
SYNAPSES AND THEN GENERATE DATA
DRIVEN CATALOGS OF SYNAPSE TYPES
FROM TWO PROTEINS YOU CAN DEFINE
THREE SYNAPSE TYPES AND 37
SUBTYPES AND THEN MAP THEM
ACROSS THE BRAIN.
NOW, THESE SUBTYPES ARE
CLASSIFIED AS I'VE MENTIONED BY
THE MOLECULAR COMPOSITION AS
WELL AS THEIR SIZE AND SHAPE
PARAMETERS ON THE BOTTOM.
BUT WHEN YOU DO THAT USING A
DATA DRIVEN APPROACH, YOU HAVE
TO ASK YOURSELF THE QUESTION,
DOES IT ACTUALLY TELL YOU
ANYTHING INFORMATIVE WHEN YOU
HAVE SO MANY SUBTYPES BEING
GENERATED?
AS SHOWN HERE IN THE MOUSE
HIPPOCAMPUS, THOSE PANELS THAT
YOU SEE ARE FOR INDIVIDUAL
SUBTYPES AND IT'S QUITE CLEAR
THAT SOME SUBTYPES ARE ENRICHED
IN PRINCIPAL AREAS OF THE
HIPPOCAMPUS, SOME ENRICHED
PARTICULARLY IN CA1, 2, CA3, AND
IN THE BOTTOM TWO PANELS YOU CAN
SEE THERE'S EVEN ARCHITECTURE
WITHIN SUB REGIONS OF THE BRAIN,
SUBTYPE 13 ON THE BOTTOM LEFT
GOES FROM STRONG WEAK ACROSS THE
CA1 STRATUM RA DIOTOM AND 24
DOES THE OPPOSITE OF THAT.
NOW, THESE MAPS THAT YOU SEE
ACROSS THE TOP SHOW YOU IN EACH
ONE OF THE PIXELS HERE THAT
THERE ARE THE COLOR CODE FOR 37
DIFFERENT SUBTYPES AND YOU'RE
SEEING THE MAIN ONE OR THE
DOMINANT SUBTYPE AND IT'S CLEAR
THERE IS THIS EXQUISITE RSZ SYN
APPEARTOME ARCHITECTURE.
LOWER, THERE'S AN INTERESTING
MAP.
WHAT WE'RE QUANTIFYING HERE IS
SYNAPSE TYPE DIVERSITY.
WE FIND 9 HIGHEST SYNAPSE TYPE
DIVERSITY IS WITHIN THOSE AREAS
OF HIGHER COGNITIVE FUNCTIONS,
CORTEX AND HIPPOCAMPUS AND MORE
BASAL STRUCTURES OF THE BRAIN
HAVE A MUCH LOWER SYNAPSE
DIVERSITY AND IF YOU LOOK INTO
THE DIFFERENT SUB REGIONS OF THE
NEOCORTEX FOR EXAMPLE AS SHOWN
HERE, THERE ARE THREE PAIRS OF
BOXES FOR SOMATOSENSORY,
VISUALIZATION AREAS AND EACH
AREA OF THE CORTEX HAS A VERY
DISTINCT LAYERING PAFT OF
DIFFERENT TYPES OF SYNAPSES,
TECTONIC MAP.
I'M NOT GOING TO SHOW YOU IN
THIS PLENGS HOW THESE
DIFFERENTIAL DISTRIBUTIONS
CORRELATE WITH PHYSIOLOGICAL
FUNCTIONS AND STRUCTURAL
CONNECTOMES, YOU CAN READ ON
THAT SUBJECT.
BUT SOME OF THE SURPRISE HAS
COME FROM SYAPTOME -- SYNAPTOME
MAPPING, HIGHLY DIVERSE, TWO
PROTEINS CAN GIVE 37 SUBTYPES,
TEN PROTEINS COULD GIVE YOU TONE
THE ELEVEN SUBTYPES WHICH IS
ACTUALLY ABOUT THE NUMBER OF
SYNAPSE PS IN THE MOUSE BRAIN
BUT IN FACT THERE'S MORE THAN
1,000 POSTSYNAPTIC PROTEINS AND
POTENTIALLY INFINITE NUMBER OF
SYNAPSE TYPES AND SUBTYPES,
CLEARLY THERE'S A LOT OF
CONSTRAINT FOR ALL SORTS OF
REASONS WHICH I WON'T DID INTO
HERE.
THE SECOND SURPRISE IS THAT THE
SYNAPTOME IS ORGANIZED DUE TO
SPATIAL OR SYNAPTOME
ARCHITECTURE AND INDIVIDUAL
DENDRITES YOU CAN SEE DIFFERENT
TYPES OF SYNAPSES ADJACENT TO
ONE ANOTHER AND DISTRIBUTED
ALONG THE LENGTH FLT DENDRITES
WITHIN THE NEURONAL DENDRITE
TREE, CELL TYPE DIFFERENCES
REGION IN THE WHOLE BRAIN
DIFFERENCES AS WELL.
BUT THEN WE WANTED TO TAKE THIS
TECHNOLOGY ONWARDS AND MAP
ACROSS THE WHOLE BRAIN OF THE
MOUSE FROM BIRTH UNTIL 18 MONTHS
TO GENERAL RAIPT THE FIRST BRAIN
WIDE QUANTIFICATION OF SYNAPSE
AND SYNAPSE TYPES ACROSS THE
LIFE SPAN OF ANY SPECIES.
AND IN EVERY BRAIN REGION AND
SUB REGION, THERE IS A
TRAJECTORY WITH AGE AND SYNAPSE
NUMBER, SYNAPSE PROTEIN
COMPOSITION, SIZE AND SHAPE AND
SYNAPSE DIVERSITY ARE
CONTINUOUSLY CHANGING.
BRAIN SYNAPTOME AND SNAP TONE
ARCHITECTURE IS CONTINUOUSLY
CHANGING ACROSS THE LIFE SPAN
BUT AT DIFFERENT RATES.
I'LL SHOW YOU A COUPLE FEATURES
OF THIS.
THESE ARE MAPS OF MOUSE BRAIN.
THE.
AT THE TOP PANEL HERE THEY'RE AT
ONE WEEK INTERVALS UP THROUGH
THREE WEEKS ACROSS TO THE FAR
RIGHT THERE AND THE BOTTOM SET
OF PANELS THEN BECOME ONE, TWO
AND THREE MONTHS OF AGE BHAVMENT
WE'RE LOOKING AT.
WHAT WE'RE LOOKING AT HERE IS
THE DIVERSITY, BLUE BEING LOW,
RED BEING HIGH.
QUITE OBVIOUSLY IN THE FIRST
COUPLE WEEKS SYNAPSE DIVERSITY
IS REALLY VERY LOW BUT AS YOU
CAN SEE IT BEGINS TO INCREASE
QUITE DRAMATICALLY AND THE
IMPORTANT THING HERE IS NOT ONLY
THAT IT IS INCREASING SYNAPSE
DIVERSITY AS THESE NEW TYPES OF
SYNAPSES POPULATE IN THE BRAIN
BUT IT IS THAT THEY GIVE EACH
AREA OF THE BRAIN ITS DISTINCT
SYNAPTOME COMPOSITION AND
SYNAPTOME ARCHITECTURE.
AND JUST ON THE OTHER END OF THE
LIFE SPAN, WE'LL HAVE A LOOK AT
AGING HERE, I'LL JUST SHOW YOU A
COUPLE THINGS.
IT'S WELL-KNOWN FROM STUDIES
FOIBLTION ODD PARTICULAR KEY
AREAS OF THE MAMMALIAN BRAIN
THAT THERE'S SYNAPSE LOSS BUT
THIS SHOWS YOU FOR THESE TWO
MARKERS HERE, THE REDUCTION IN
SYNAPSE NUMBER THAT OCCURS FOR
THOSE IN THESE DIFFERENT BRAIN
AREAS THAT WE'VE EXAMINED.
OVERALL THERE'S A GENERAL LOSS.
BUT WHEN YOU NOW LOOK AT THE
SYNAPSE SUBTYPES AND I'M JUST
SHOWING YOU TWO HEAT MAPS HERE
FOR FOUR SUBTYPES, LET ME ORIENT
YOU ON THESE HEAT MAPS.
THE VERTICAL AXIS IS ABOUT 100
OR SO DIFFERENT BRAIN REGIONS
AND HORIZONTALLY WE'RE LOOKING
AT THEIR LIFE SPAN TRAJECTORY
FROM THE FIRST DEAF LIFE ON THE
LEFT DISPOF 08 MONTHS ON THE
RIGHT.
TWO HEAT MAPS ON THE LEFT IN
GENERAL THE YELLOW WHICH IS THE
HIGHEST DENSITY FOR THESE
PARTICULAR SUBTYPES IS GENERALLY
PRETTY HIGH IN ABOUT THE SECOND
AND THIRD COLUMNS FROM THAT AND
THEN THEY'RE GENERALLY BECOME
REDUCED AS YOU GET OLDER.
THESE ARE EAJ SENSITIVE, THESE
ARE SUBTYPES THAT PEAK EARLY IN
LIFE AND GENERALLY BECOME LOST
A'S THE ANIMAL GETS OLDER.
BUT ON THE OTHER HAND THESE
SECOND THE LAST TWO THIRD AND
FOURTH HEAT MAPS YOU CAN SEE ARE
THE OPPOSITE OF THAT.
THESE ARE EAJ RESILIENT
SYNAPSES, THESE ARE SUBTYPES OF
SYNAPSES THAT DEFY THE AGING
PROCESS AND REMAIN ROBUSTING AND
ARE NOT LOST AS YOU GET OLDER.
SO CLEARLY A VERY INTERESTING
QUESTION IS TO ASK WHAT'S
DIFFERENT ABOUT THOSE SYNAPSES?
I'LL COME BACK TO THAT LATER ON
AND SHOW YOU SOMETHING QUITE
INTERESTING ON THAT SUBJECT.
BUT I WOULD JUST LIKE TO
SPECULATE FOR A MOMENT, FROM
HUMAN STUDIES IN PARTICULAR,
THERE'S BEEN A VERY CLEAR
DICHOTOMY AND COGNITIVE FUNCTION
WITH AGING WHERE THERE IS A
REDUCTION AS SHOWN IN THESE
LEFT-HAND GRAPHS WITH AGING
HUMANS OF WHAT'S KNOWN AS FLUID
COGNITION WHICH ESSENTIALLY IS
PROCESSING SPEED, WORKING MEARM,
ATTENTION, EXECUTIVE FUNCTION,
AND LEARNING EFFICIENCY.
BUT THAT IS IN CONTRAST TO
ANOTHER TYPE OF COGNITION KNOWN
AS CRYSTALLIZED COGNITION WHICH
IS STABLE WITH AGING, THAT'S
ACQUIRED KNOWLEDGE AND REALLY
CORE CAPACITIES SUCH AS LANGUAGE
AND IT'S AN INTERESTING THING TO
SPECULATE AND PERHAPS SOMETHING
WE COULD ULTIMATELY TEST OF
THESE DIFFERENT SYNAPSES MIGHT
BE ASSIGNED TO THESE DIFFERENT
TYPES OF FUNCTIONS.
I'M GOING TO CHANGE SUBJECTS
HERE BECAUSE WHAT I'VE SHOWN YOU
IS THE MOLECULAR COMPOSITION,
STALT I CAN COMPOSITION, BUT
PROTEINS HAVE A LIFETIME.
THEY TURN OVER.
SO PROTEINS WITH LONG LIFETIMES
HAVE A VERY RAPID TURNOVER RATE.
SORRY, JUST BENEATH THAT, LONG
LIFETIME, SO SLOW PROTEIN
TURNOVER RATE AND THAT'S AN
IMPORTANT THING BECAUSE THAT
MEANS THERE'S A SLOW REMOVAL OF
MODIFIED OR DAMAGED PROTEINS AND
SLOW PROTEOSTASIS MEANS SLOW
HOMOEOSTASIS AND SLOW
ADAPTATION.
SO THE QUESTION IS, DO SYNAPSES
HAVE DIFFERENT PROTEIN
LIFETIMES?
NOW, THE LIFETIME OF ENDOGENOUS
SYNAPTIC PROTEINS HAS NOT BEEN
STUDIED AT SINGLE SYNAPSE
RESOLUTION AND CERTAINLY NOT AT
A BRAIN WIDE SCIEL WE DEVELOPED
A METHAMPHETAMINE ODD TO DO THAT
WHICH IS REPORTED IN THIS PAPER
ON NILE ARC IEFDZ LED BY Ph.D.
STUDENT AND WHAT WE'VE USED HERE
IS THE PALO TAG ATTACHED TO THE
PSD95 PROTEIN AND ON THE LEFT
YOU CAN SEE A SCHEMATIC
THREE-DIMENSIONAL STRUCTURE OF
THE HALO PHAGO WHICH IS
ESSENTIALLY CONTAINS -- HALO TAG
WHICH ESSENTIALLY CONTAINS A
BINDING POCKET INTO CL A SMALL
MOLECULE FORMS A COVALENT BOND
AND THUS PERMANENTLY BINDING TO
THE HALO TAG PROTEIN DOMAIN.
THE BEAUTY OF THIS IS THAT YOU
CAN COUPLE TO THAT HALO TAG
LIGAND ANYTHING YOU LIKE SUCH
THAT IF YOU WERE TO INJECT THIS
INTO THE MOUSE AND LABEL PS95
WITH HALO TAG ON IT, YOU'LL GET
IRREVERSIBLE LAG EXPLENGT SEE
THE DYNAMICS AS IT'S LOST
THROUGHOUT TIME.
HERE IS A SCREMATIC PRESENTATION
WHAT YOU MIGHT LOOK FOR.
TWO ROSE.
TOP ONE, TYPE OF SYNAPSE WHERE
PROTEINS HAVE BEEN A SHORT
LIFETIME AND ONE AT THE BOTTOM
IS A LONG PROTEIN LIFETIME
SYNAPSE.
THEY CONTAIN PSD95 IN THE
POSTSYNAPTIC REGION OF THE
SYNAPSE.
WHERE THE RED BAR SAYS HALO
INJECTION, ATTACHED TO HALO, WE
INJECT THE MICE AND CAN LABEL
ALL THE PROTEIN.
THE TOP SYNAPSE TURNS OVER THE
PROTEIN VERY QUICKLY, SYNAPSE
PROTEIN HAS A SHORT LIFETIME SO
IT DISAPPEARS RAPIDLY UNLIKE
THAT AT THE BOTTOM WHERE IT HAS
A SLOW PROTEIN TURNOVER AND A
LONG PROTEIN LIFETIME WHERE IT
IS RETAINED SO THEREFORE SIMPLY
BY INJECTING THE MICE WITH THIS
LIE LIGAND AND USUALLY -- AND
USING PIPELINE MAPPING, WE CAN
DO SANG HE WILL RESOURCE MAP OF
PROTEIN TURNOVER IN THE MOUSE
BRAIN.
I'LL SHOW YOU A COUPLE CLOSEUPS
THAT ILLUSTRATE FOR YOU THAT THE
REASONS ARE TURNING OVER IN
SYNAPSES.
ON THE TOP LEFT IF YOU NEED A
CLOSE-UP OF THE MOTOR CORTEX FOR
ANIMALS THAT HAVE BEEN INJECTED
WITH ONE DYE ON DAY ZERO AND
THREE DAYS LATER WITH ANOTHER
DYE AND ONE BEING RED AND THE
OTHER GREEN AND HENCE YOU CAN
SEE THE MERGED SIGNAL AND THAT
MERGED SIGNAL TELLS YOU THAT THE
PROTEINS ARE TURNING OVER IN
THOSE SYNAPSES.
IN COLLABORATION WITH ANTHONY
HOLTMART'S GROUP IN GENEVA HE
IMAGED WITH LIVE MICE IN LAYER
TWO OF THE CORTEX THE TURNOVER
THE SYNAPTIC PROTEINS USING THE
HALO TAG OVER A PERIOD OF A
MONTH OR MORE AND HE COULD
INDIVIDUALLY SEE THE DENDRITIC
SPINES THERE AS SHOWN IN THE TOP
AND THE ILLUSTRATED AT THE
BOTTOM SET OF THREE PANELS
LABELED WITH HALO TAG AND THE
INDIVIDUAL SYNAPSES ON THESE
DENDRITES HAVE DIFFERENT
LIFETIME AS SHOWN ON THAT GRAPH
ON THE RIGHT, SOME SYNAPSES LOSE
THE PROTEIN OR HAVE A VERY SHORT
LIFETIME AND OTHERS HAVE A LONG
LIFETIME.
SO HERE ARE SOME EXAMPLES OF I
AM NOODGES THE BRAIN, EACH
COLUMN IS A PARTICULAR DAY AFTER
INJECTION, IF YOU LOOK AT THE
TOP AT THE WHOLE BRAIN YOU'LL
SEE VERY CHARACTERISTIC LABELING
OF 95 AT DAY ZERO.
BY DAY 14 YOU CAN SEE THAT
ESSENTIAL ALMOST ALL ALL THE --
IS GONE AND I MUST STOP AND
PAUSE FOR A SECOND BECAUSE I
THINK ITS TRULY ASTONISHING WHEN
YOU HAVE THIS AMAZE BEING SYN
APPEAR TOMORROW ARCHITECTURE.
BRAIN -- SYNAPTOME ARCHITECTURE
OF THE BRAIN, THAT IT'S BEING
REBUILT EVERY COUPLE WEEKS AND
THE BRAIN HAS THIS REMARKABLE
ABILITY TO DO THAT.
SOME OF THE SIGNAL REMAINS IT
SOME OF THESE AREAS AND SOME OF
THESE SYNAPSES HAVE A VERY LONG
LIFETIME.
HERE WE'VE MAPPED THE AVERAGE
HALF-LIFE SYNAPTIC PROTEINS IN
THESE DIFFERENT AREAS OF THE
BRAIN AND THEY DIFFER BY TENFOLD
IN THESE DIFFERENT REEXPWRONS
QUITE NOTICEABLY YOU'LL --
REEXPWRONS QUITE NOTICEABLY
YOU'LL SEE THE SUPER SUPERFICIAL
LAYERS OF THE CORTEX HAVE A VERY
SLOW TURNOVER OF PROTEIN AND
VERY LONG PROTEIN LIFETIME.
IF YOU LOOK WITHINS NEURONONTZ
CA LEE REGION YOU CAN SEE ON
THIS GRAPH ON THE Y AXIS LOOKING
AT THE ESSENTIAL PROTEIN
LIFETIME TO DISTAL DENDRITES TO
THE RIGHT YOU CAN SEE THERE'S AN
OVERALL GRADIENT THERE SUCH AS
MORE DISTAL SYNAPSES HAVE A
LONGER PROTEIN LIFETIME THAN
THOSE THAT ARE MORE PROXIMAL.
AND ONE OF THE QUESTIONS WE WANT
TO ASK IS WHETHER OR NOT THESE
COMPOSITIONAL SUBTYPES AND
THIEPS WE DEFINED USING THESE
GFP AND ORANGE LABELS ARE IN ANY
WAY RELATED TO THE PROTEIN
LIFETIMES.
COULD IT BE THAT SOME OF THESE
SUBTYPES HAVE PARTICULAR
COMPOSITIONS THAT HAVE DIFFERENT
LIFETIMES?
WE DID THIS BY CROSSING TO MAKE
A TRIPLE LABELED MOUSE WITH 95
SPAITION GFP AND ORANGE WHICH
GAVE UPS THE SUBTYPE
CLASSIFICATION AND ALSO HAD THE
PSA95 HALO TAG SHES PROTEIN
LIFETIME.
AS SHOWN IN THESE GRAPHS BELOW
HERE, THE GRAPH BELOW, WE'VE
RANKED THE SUBTYPES ACHORD TO GO
THEIR PROTEIN LIFETIME --
ACCORDING TO THEIR PROTEIN
LIFETIME.
IT'S CLEAR THERE ARE SOME
SUBTYPES THOSE SHOWN ON THE
LEFT-HAND SIDE WHICH ARE LONG
PROTEIN SYNAPSE SUBTYPES AND FAR
RIGHT THERE ARE THOSE WITH SHORT
PROTEIN LIFETIMES.
SO IN OTHER WORDS, THESE
SUBTYPES OF SYNAPSES WITH
DIFFERENT LIFETIMES ARE REALLY
FUNDAMENTAL BUILDING BLOCKS OF
THE SYNAPTOME AND THEY'RE
POPULATING DIFFERENT AREAS OF
THE BRAIN.
LET'S GET BACK TO THIS PICTURE
THAT I SHOWED YOU EARLIER ABOUT
THESE VERY INTERESTING DIFFERENT
SUBTYPES OF SYNAPSE THAT IS ARE
ON ONE HAND AGE SENSITIVE AND
AGE RESISTANT AND IT TURNS OUT
IT WOULD BE VERY NICE THAT THE
PROTEINS THAT HAVE THE SHORT
PROTEIN LIFETIME ARE THE AGE
SENSITIVE ONES AND IN OTHER
WORDS RAPIDLY TURN OAFERG,
RAPIDLY REMODELING AND THOSE ARE
RESISTANT TO OLD AGE HAVE THE
ONES WITH LONG PROTEIN LIFETIME.
WE THINK THERE'S SOMETHING VERY
SPECIAL ABOUT THAT, THAT IS TO
SAY APPEARS THE ANIMAL IS AGING,
SLOW SYNAPSE RS PREZIL YENT TO
WHATEVER LOSSES MIGHT OCCUR.
SO WE'VE ALSO DONE A STUDY
ACROSS THE LIFE SPAN AT THE
PROTEIN LIFETIME AND SEE HERE
THREE BRAIN MAPS AT THREE WEEKS,
THREE MONTHS AND 128 MONTHS.
THERE'S A VERY -- AND 18 MONTHS.
THERE'S A VERY CLEAR TREAND
CROSS THE LIFE SPAN WHERE THE
PROTEIN LIFETIME INCREASES
AGGRESSIVELY GENERALLY IN ALL
AREAS OF THE BRAIN APPEARS THE
ANIMAL GETS OLDER.
SO IN ESSENCE WHAT THIS MEANS
AND THIS IS CONSISTENT WITH
OTHER STUDIES THAT HAVE THIS
SEASON THAT PROTO PROTEOSTASIS
SLOWS DOWN AS ANIMALS AGE.
SO I WOULD JUST LIKE TO FINISH
HERE BY REALLY EMPHASIZING THE
IMPORTANCE OF WHAT ARE NOW THREE
PARAMETERS, QUANTITY OF THE
PROTEIN, THE MORPHOLOGY THAT IS
REALLY THE NANO ARCHITECTURE OF
THE PROTEINS AND THE PROTEIN
LIFETIME AND EACH OF THESE VARY
BETWEEN SYNAPSES AND THESE THREE
FUNDAMENTAL COMPONENTS REALLY
ARE KEY TO THE SYNAPSE DIVERSITY
OF THE NERVOUS SYSTEM AND THESE
DIFFERENT DIVERSE SYNAPSES ARE
THEN DISTRIBUTED AND ORGANIZED
IN THE BRAIN TO GIVE THIS
SYNAPTOME ARCHITECTURE AND
UNDOUBTEDLY UNDERLIE KEY
FUNCTIONS WHETHER THEY BE
SYNAPTIC TRANSFORMATION MISSION
AND SYNAPTIC PLASTICITY AND EVEN
MEMORY FUNCTIONS.
WITH THAT I'LL FINISH AND THANK
YOU FOR YOUR TIME.
BUT I WANT TO TELL YOU WHO HAS
REALLY DONE THIS MAIN WORK, I'VE
MENTIONED THAT ADIDA GUVALADE
HAS LED THIS AND -- PLAYED A KEY
ROLE IN THE DESIGN AND MICE AND
RICKY IS SPECIAL CONSIDERATION
GIVEN THAT GIVEN THAT HE'S
DEVELOPED THIS SPECIAL IMAGING
PIPELINE FOR THE SYNAPTOME
MAPPING.
THANK YOU VERY MUCH.
>> I WOULD LIKE TO THANK ALL THE
SPEAKERS FOR THEIR WONDERFUL
TALKS.
AND NOW WE HAVE SOME TIME FOR
QUESTIONS FOR FOLKS LISTENING
IN.
PLEASE USE THE Q & A TAB ON THE
RIGHT SIDE PANEL OF YOUR BROWSER
TO ADD QUESTIONS THAT I WILL
SUBSEQUENTLY FOAPS THE
SPEAKERS -- POSE TO THE
SPEAKERS.
LET'S SEE.
FIRST OF ALL, TWO QUESTIONS FOR
SETH.
FIRST, DO YOU EXPECT THESE
CHANGES IN SYNAPSE SUBTYPES OVER
AGE TO BE DRIVEN BY EPIGENETIC
CHANGED OR MORE LOCAL CHANGES IN
THE SYNAPTIC MOLECULAR NETWORK?
>> THAT'S A GOOD QUESTION ABOUT
EPIGENETIC CHANGED.
WE CERTAINLY KNOW THAT THERE ARE
TRANSCRIPTIONAL CHANGES
INVOLVED.
UP UNTIL THE FIRST THREE MONTHS
OF AGE.
BUT THESE LIFETIME CHANGES THAT
OCCUR N OUR SCIENCE PAPER WE
SHOWED THAT FROM THREE MONTHS OF
AGE OUT TO ABOUT 18 MONTHS IT'S
A SLOW CS SET OF CHANGES
AFFECTING SOME OF THE SUBTYPES.
I THINK THERE'S GOING TO BE
POST-TRANSCRIPTIONAL MECHANISM
IN ADDITION TO TRANSCRIPTIONAL
REGULATION, FOR EXAMPLE, THEAZ
EFFECTS ON TRANSLATION PROTEIN
LIFETIME ARE GOING TO BE -- I
THINK ALL OF THESE FACTORS WILL
CONTRIBUTE IN SOME WAY TO THE
SYNAPTOME ARCHITECTURE AND AN
IMPORTANT POINT I WANT TO
EMPHASIZE HERE IS THAT WHEN ONE
UNDERSTANDS THESE VERY
ELEMENTARY MOLECULAR PRINCIPLES
SUCH AS REGULATION OF PROMOTOME
FOR EXAMPLE IT CAN EXPLAIN WHY
MUTATIONS LEAD TO CHANGES IN THE
SYNAPTIC ARCHITECTURE WHICH IN
TURN PRODUCE BEHAVIORAL
DISPLAING WO CAN I QUICKLY
FOLLOW UP.
>> CAN I QUICKLY FOLLOW UP WITH
A SLIGHTLY DIFFERENT QUESTION,
WHICH IS DO YOU SEE MORE
DIVERSITY IN KIND OF THE
PRESYNAPTIC OR POSTSYNAPTIC KIND
OF COMPARTMENTS IN TRMS OF BOTH
DIVERSITY AND LIFETIME?
>> NO, WE FOCUSED MOST OF OUR
ATTENTION ON THE POSTSYNAPTIC
SIDE.
WE CERTAINLY HAVE DONE SOME
LIMITED STUDIES ON PRESYNAPTIC
TERMINALS AND THERE'S CERTAINLY
A LOT OF DIVERSITY THERE TOO.
I THINK THESE ARE GENERAL
PRINCIPLES THAT APPLY BOTH TO
PRE AND TO POST, BUT THERE'S
OBVIOUSLY A LOT OF VERY KEY
SPECIFIC QUESTIONS TO LOOK AT IN
BOTH CIRCUMSTANCES.
>> EXCELLENT.
IS THERE A PLAN TO COMBINE ALL
CONNECT TOMORROW SYNAPTOME
EFFORTS IN THE END OR WILL
GENALIA STAY SEPARATE SOME
PLATFORMS LIKE FLY WIRE?
PERHAPS CLAIRE CAN START WITH
THAT, ALTHOUGH UNFORTUNATELY
MICHAEL IS NOT WITH US FOR THE Q
& A PORTION.
>> IT'S MY DREAM THAT SCIENCE
WILL BECOME MORE AND MORE OPEN
SO WE'VE TRIED TO SET AN EXAMPLE
THERE, AND WE HOPE THAT OTHER
PEOPLE WILL BE INTERESTED IN
COLLABORATING AS MUCH AS
POSSIBLE.
SO I'VE REALLY ENJOYED HOW MUCH
THE PROJECT BETWEEN GENELIA AND
FLY WIRE HAS BEEN TRACKED AND I
HOPE TM CONTINUE TO GROAVMENT
ADDING IN SYNAPTOME I'LL LET
SETH SPEAK TO THAT MAYBE.
>> I WOULD JUST LIKE TO SAY ONE
THING ABOUT IT, AND I THINK IT'S
VERY OBVIOUS TO EVERYBODY, WHEN
ONE DESCRIBES THE CONNECT OHM
AND SOME WHO HAVE DONE ALREADY
HERE TODAY AND MANY OTHERS HAVE
DONE TOO, IT'S USUALLY IN TERMS
OF THE PHYSICAL LOCATION OF THE
SYNAPSES WITH RESPECT TO THE
DENDRITES AND AXONAL PROJECTIONS
AND OBVIOUSLY THE MOLECULAR
COMPOSITION MAP OF ALL OF THOSE
SYNAPSE AND IS CLEARLY THESE TWO
THINGS NEED TO BE INTEGRATED AND
THERE'S A WHOLE LOT OF TECHNICAL
CHALLENGES ASSOCIATED WITH THAT
AND SOME OF WHICH WE'RE WORKING
ON BUT OBVIOUSLY THESE ALL HAVE
TO BE PUT TOGETHER AT SOME POINT
BECAUSE IF YOU DON'T HAVE THE
MOLECULES IN YOUR CONNECT OHM --
IN YOUR CONNECTOME, YOU DON'T
HAVE THE FUNCTION.
>> I GUESS I WOULD ADD, I WOULD
ECHO CLAIRE AND KIND OF ADD THAT
IN GENERAL, THESE CONNECTOMICS
DATA SETS ARE BEING PUT OUT TO
THE PUBLIC AT SOME POINT SUCH
THAT PEOPLE CAN, YOU KNOW, THAT
THE DATASETS THEMSELVES PROVIDE
AN OPPORTUNITY TO FUEL FURTHER
KIND OF SCIENCE AND DISCOVERY,
BUT THERE IS A CHALLENGE OF KIND
OF INTEGRATING DIFFERENT
MODALITIES, FOR EXAMPLE, IN
DIFFERENT SAMPLES.
BUT I THINK THIS IS HOPEFULLY
FOOD FOR THE FUTURE.
HI, CLAIRE, IS THERE ANY PLAN TO
PROOFREAD THE AUTO-DETECTED
SYNAPSE INSIDE FLY WIRE?
>> THAT WOULD BE GREAT.
RIGHT NOW, NOT IMMEDIATELY.
BUT WE ARE GOING TO HAVE SUCH A
NUMBER OF HUMAN RESOURCES THAT
AFTER THE NEURONS HAVE BEEN
PROOFREAD, IT MIGHT MAKE SENSE
TO TURN THE WORLD'S ATTENTION TO
THE SYNAPSES.
ZOOG OBJECTIVE.
>> OKAY.
THE SYNAPSE PREDICTION, THE KIND
OF PRECISION RECALL OF THE
AUTO-DETECTED SYNAPSES IS FAIRLY
GOOD, NOT PERFECT, OBVIOUSLY,
BUT --
>> IT IS, BUT THE SHEER NUMBERS
MAKE IT ALMOST IMPOSSIBLE FOR
HUMAN INTERVENTION, BUT THERE
COULD BE HUMAN INTERVENTION AT
SOME KIND OF SCALE.
>> SO A QUESTION FOR BEN.
IT'S FASCINATING THAT THE MOTOR
BEHAVIOR STAYS THE SAME DURING
C. ELEGANS MATURATION WHILE
MOTOR NEURONS ARE REMODELED.
COULD YOU MAKE ANY PREDICTIONS
ABOUT SYSTEM WHERE THE BEHAVIOR
ACTUALLY CHANGES BUT STILL USES
THE SAME MOTOR NEURONS?
FOR EXAMPLE, FLY LARVA AND
ADULTS SHARE MANY OF THE SAME
MOTOR NEURONS FOR FEEDING BUT
COMPLETELY DIFFERENT MOTOR
NEURONS.
DO YOU EXPECT MODELING
MECHANISMS YOU FOUND COULD APPLY
IN SYSTEMS LIKE THAT?
>> YEAH.
SO I DON'T KNOW MUCH ABOUT THE
TIMING OF DROSOPHILA, BUT
DROSOPHILA CHEATS AND THEY GO
INTO THE PEOPLE STAGE.
COINCIDE WITH THAT.
IN WHICH CASE IT MIGHT BE KIND
OF LESS CONSTRAINED IN
MAINTAINING FUNCTION.
>> OKAY.
FOR STEPHANIE.
BEYOND THE MECHANOSENSORY
NEURONS YOU FOUND, HOW
SEGREGATED DO YOU THINK THE
INFORMATION WILL REMAIN OR LIKE
WHEN DOES IT GET INTEGRATED TO
PROPAGATE DOWN TO KIND OF
DOWNSTREAM CIRCUITS, GROOMING
CIRCUITS AND KIND OF ULTIMATELY
MOTOR NEURONS THAT ARE EXECUTING
THESE GROOMING BEHAVIORS?
>> YEAH, WE HAVE ACTUALLY
RECONSTRUCTED THE ENTIRE NETWORK
IN THE BRAIN SO IT BASICALLY
GOES THROUGH ABOUT TWO LAYERS OF
NEURONS IN THE BRAIN AND THEN ON
THE VENTRAL NERVOUS SYSTEM,
WHICH WE ALSO HAVE STARTED IN
WAYS WHICH I HAVEN'T TALKED
ABOUT TODAY, AND A LOT OF THE
DESCENDING NEURONS ARE DIRECTLY
CONNECTED TO MOTOR OUTPUTS.
SO YEAH.
>> EXCELLENT.
WELL, I THINK I HAVEN'T ASKED
THIS QUESTION.
THIS IS FOR SETH.
WHAT ARE SOME OF THE MAJOR
BARRIERS RELATING SYNAPTOME
TOAMS AS DESCRIBED TO YOU TO
MOLECULARLY AND ANATOMICALLY
DESCRIBED CIRCUITS AT SOME
CELLULAR SCALE IN SOME OTHER
TALKS?
>> THE QUESTION THAT WE ARE
WORKING ON AT THE MOMENT WHICH
MAY ADDRESS THAT IS TO DEVELOP
CELL TYPE SPECIFIC WAYS OF DOING
SYNAPTOME LABELING AND FOR
EXAMPLE USING PRELOX P METHODS
WHERE YOU CAN CONDITIONALLY
TAPPING THE MOLECULES.
THAT WORKS QUITE NICELY.
WHAT WE NEED TO DO AND AGAIN I
THINK THERE'S A LOT OF
TECHNOLOGY IN THE CONNECTOMICS
WORLD THAT -- IN THE
CONNECTOMICS WORLD THAT HASN'T
BEEN EXPLORED, RS THE LONG
RANGE, I WOULD BE INTERESTED TO
TALK WITH PEOPLE ABOUT THAT
SHOULD THEY HAVE AN INTEREST IN
DOING THAT.
>> AND THANKS FOR THE GREAT
TALKS.
I HAVE A QUESTION FOR THE GROUP.
WHAT DO YOU SEE AS A
RATE-LIMITING STEPS IN TERMS OF
SCALING PROOFREAD TO GO LARGER
VOLUMES, FOR EXAMPLE, MAMMALIAN
BRAINS OR MULTIPLE SMALL BRAINS
OF THE SAME SPECIES SAMPLED
UNDER DIFFERENT CONDITIONS OR
GENETIC BACKGROUNDS?
MAYBE CLAIRE CAN START.
>> CAN I TAKE THAT ONE?
>> SURE, YES.
>> I THINK THAT IT WOULD BE
GREAT TO HAVE MORE AUTOMATED
ERROR DETECTION USING HEURISTICS
LIKE BEING ABLE TO MAKE
PREDICTIONS ABOUT AXONS VERSUS
DEN DRIETD AND SAY, WELL, THIS
IS A PIECE OF AXON STUCK TO A
DEN TRANSMIT I CAN TREE SO HUMAN
PLEASE GO LOOK THERE, OR, YOU
KNOW, RS -- SUPPOSED TO BE FROM
THICK TO THIN, SO WHY IS THIS
BRANCH THICK?
SO OBVIOUSLY THE BETTER THE
ALGORITHMS BECOME, THEN THE LESS
HUMAN INVOLVEMENT IS NEEDED BUT
AS YOU SCALE UP, ACTUAL ACTUALLY
THE MORE HUMAN INVOLVEMENT IS
NEEDED BUT WE NEED MORE TOOLS TO
MAKE HUMANS' JOBS EASIER.
>> YEAH, I WOULD ECHO THAT.
THERE'S BEEN SUBSTANTIAL
PROGRESS IN TERMS OF LIKE
JAINTING THESE TYPES OF DATA --
GENERATING THESE TYPES OF DATA
AND THEN SUBSEQUENTLY KIND OF
DOING THE INITIAL KIND OF
SEGMENTATION AND THINGS LIKE
SYNAPSE PREDICTION BUT KIND OF
THE BOTTLENECK AT THIS POINT AND
CONTINUES TO BE KIND OF THE
PROOFREADING AND KIND OF THE
DETEKS OF THOSE ERRORS AND AS
THE AUTOMATIC ALTHOUGH GOA
RHYTHMS GET BETTER AND BETTER
FINDING THOSE ALGORITHMS GETS
HARDER AND HARD AND HER THAT'S
WHERE THE TIME IS GOING TO BE
SPENT BY HUMAN EXPERTS ALTHOUGH
THIS ALSO DEPENDS THE QUESTIONS
THAT PEOPLE ARE TRYING TO
ADDRESS USING THESE CONNECT
OHMS, LIKE HOW TOLERANT THEY TO
KIND OF ERRORS AND THINGS LIKE
SEGMENTATION OR SYNAPSE
PREDICTION.
OKAY.
MAYBE ONE FINAL -- OH, MAYBE TWO
FINAL QUESTIONS.
THIS IS TO DR. GRANT.
WHAT DO YOU THINK, OR DO YOU
THINK LOCATION INSTRUCTS
DIFFERENT SYNAPSE SUBTYPES?
HOW DO A NEURON -- HOW DOES A
NEURON KNOW, FOR EXAMPLE, THAT
IT WANTS SYNAPSE TYPE A ON FOR
EXAMPLE A DISTAL DENDRITE OR
TYPE B ON A PROXIMAL DENDRITE,
FOR EXAMPLE?
>> WELL, I DON'T WISH TO BE
FUSSY ABOUT THIS, BUT NEURONS
DON'T WANT ANYTHING.
>> SURE.
>>
>> BUT IF YOU WERE TO ASK THE
QUESTION HOW DO THE MOLECULES
GET THERE AND HOW THEY POPULATE
THOSE AND MAKE DIFFERENT TYPES
OF SYNAPSES, FOR EXAMPLE, WE SEE
THERE ARE CLEAR GRADIENTS ALONG
THE LECT OF THE DENDRITE --
LENGTH OF THE DENDRITE, PROTEIN
PROTEIN LIFETIME AND PROTEIN
PROTEIN COMPOSITION AND I EXPECT
THAT WOULD BE DID NAITD BY VERY
NINTD AL CELL BIOLOGICAL
PRINCIPAL PRINCIPLES OF TRACK
AND GO SO ON AND BY DEFAULT
GENERATE DEN CITE DIVERSITY.
ALSO WE SEE PROGRAM TELL CELL
THAT DIFFERENT INPUTS ALONG THAT
DIFFERENT, YOU CAN SEE THERE ARE
QUITE DIFFERENT TYPES OF
SYNAPSES THERE AND WITHIN THE
THALAMUS ON AN INDIVIDUAL
DENDRITE YOU CAN SHOW THAT THERE
ARE ADJACENT SYNAPSES WHICH HAVE
DIFFERENT INPUTS ONE FROM THE
CORTEX AND ANOTHER FROM THE
BRAINSTEM, FOR EXAMPLE, AND THEY
HAVE REALLY QUITE DIFFERENT
SYNAPSE PROTEIN COMPOSITIONS AT
THE POSTSYNAPTIC SIDE.
RSZ CONSTRUCTIVE EFFECTS IN
PRESYNAPTIC PROTEINS HAVE BEEN
DEMONSTRATED.
IT WILL BE THESE SORTS OF
MECHANISMS THAT WILL GIVE YOU
THE ULTIMATE SYNAPTOME
ARCHITECTURE.
>> AND ONE FINAL QUESTION, IT
LOOKS LIKE IN THE Q & A, MAY I
ASK IF THERE ARE PLANS TO
IMPROVE SEGMENTATION/SYNAPSE
PREDICTION OF CELLS FOR EXAMPLE
WITH DARK CYTOSOL POSSIBLE
EXAMPLES INCLUDE GLIA IN THE EM
DATA?
CLAIRE, ARE YOU --
>> YES, WE ARE DOING THAT,
ACTUALLY.
SO WE'VE GENERATED NEW GROUND
TRUTH THAT INCLUDES CELLS WITH
DARKER CYTOSOL AND WE HAVE
RESEGMENTED AND REALIGNED THE
SAME FLY BRAIN DATASET THAT FLY
WIRE ALREADY USES, AND THERE
WILL BE A BIG EFFORT TO FIGURE
OUT HOW TO INTEGRATE THAT WITH
ALL OF THE HUMAN EFFORT THAT HAS
GONE INTO THE CURRENT DATA SET.
SO WE'LL HAVE TO TALK ABOUT HOW
TO PHASE IN THE NEXT
SEGMENTATION, BUT WE HOPE THAT'S
COMING, AND I DO NOT YET HAVE AN
ANSWER FOR YOU ABOUT HOW MUCH
BETTER OR DARK CELLS MIGHT BE
SEGMENTED, BUT WE HOPE THAT THEY
WILL BE.
BUT WE OURSELVES ARE NOT
DIRECTLY INVOLVED IN ANY SYNAPSE
PREDICTION EFFORT AT THE MOMENT
FOR FLY WIRE.
>> WELL, I THINK THAT BRINGS US
TO THE END OF THE Q & A SESSION
AND THE SYMPOSIUM SESSION MORE
BROADLY SPEAKING.
THANK YOU AGAIN FOR EVERYONE
THAT HAS ATTENDED, AND THANKS
EVERYONE FOR THEIR QUESTIONS AND
THE WONDERFUL SERIES OF TALKS.