>> IT'S NICE TO SEE SO MANY PEOPLE HERE AND WELCOME TO AUDACIOUS GOALS. WE STARTED THIS YEARS AGO, IN ORDER TO GENERATE CAPACITY TO THE RETINA AND VISION. WE HAD A BUSY YEAR FOR THE AGI, WE WANT TO PUT IN A PLUG FOR THE SPONSOR FOR TODAY. THE AGI, THIS IS THE SECOND YEAR OF FUNDING, THE FIRST YEAR IN 2015 WENT TO FIVE COLLABORATIVE PROJECTS TO DEVELOP IMAGING TOOLS. THE CONCEPT OF AGI IS TO RESTORE FUNCTIONALITY OR IN FACT RESTORE CELLS IN THE RETINA, PARTICULARLY THE PHOTORECEPTOR CELLS AND THE RETINAL GANGLIAN CELLS THIS INTO HUMAN PATIENTS. IN ORDER TO DO THAT WE NEED A WAY OF LOOKING AT THOSE CELLS TO IMAGE THOSE CELLS. SIMILAR TO THE ADVANCES THAT HAVE BEEN MADE IN RETINAL DISEASES THROUGH OPTICAL COHERENCE TOMOGRAPHY, WE NEED STEPS THAT WILL GET US DOWN TO THE INDIVIDUAL CELL LEVEL AND THAT WAS THE FIRST FUNDING TRENCH OF AGI IN 2015. THE SECOND FUNDING WHICH OCCURRED IN AUGUST OF THIS YEAR 2016 IS TWO IDENTIFY BIOLOGIC FACTORS THAT EFFECT NEUROREGENERATION IN THE RETINA. JUST A COUPLE OF POINTS YOU MAY WANT TO KNOW ABOUT, THERE IS A VERY INTERESTING ARTICLE IN THE JOURNAL OF NEUROSCIENCE I THINK IT WAS OCTOBER 2016, AUTHORED BY MIKE CRYER AT YALE AND CAROL MASON ON COLUMBIAOT OUTPUT OF AN AGI WORKING GROUP, THE TOPIC WAS TO RECONNECT THE EYE TO THE BRAIN. NOT A SIMPLE CONCEPT BUT THESE TWO PEOPLE HAVE LAID OUT ALONG WITH THE BACKING OF THE WORKING GROUP, HAVE LAID OUT SOME REALLY INTERESTING CONCEPTS, IMPORTANT THINGS TO KNOW ABOUT IN THIS VENTURE. THIS LAST YEAR OR A YEAR AGO AT THE ARVO MEETING THERE WERE TWO AGI EVENTS, ONE ON REPLACING RETINAL GANG LIAISON AN CELLS AND THE SECTD WAS A TOWN HALL IN HOW ONE INTERFACES ALL THE WONDERFUL PRISTINE WONDERFUL BASIC DISCOVERY BIOLOGY WE HAVE ANCHORED IN THE RETINA AND HOW ONE ANCHORS THAT TO HUMAN DISEASE ULTIMATELY TO HAVE THE AGI EXTEND TO THE HUMAN DISEASE CONDITION BUT ONE NEEDS TO HAVE A SET OF CLINICIANS WHO ARE RECEPTIVE TO THE IDEAS AND CAN HELP GIED THE PROCESSES THAT WILL UNDERPIN THIS SO THAT WE KNOW WHAT THE TARGETS ARE THAT WE'RE TRYING TO GET INTO. BUT ENOUGH ABOUT WHERE WE HAVE BEEN, I'D NOW LIKE LIKE TO INTRODUCE ANDIE HUBERMAN, ANDY IS AN ASSOCIATE PROFESSOR AT STANFORD, HE'S WITH NEUROBIOLOGY AND OPHTHALMOLOGY AND ALSO WITH THE BIO-X PROGRAM WHICH IS AN INTRIGUING INTERDISCIPLINARY PROGRAM, ENTREPRENEURIAL PROGRAM AT STANFORD. HIS TRAINING, Ph.D. FROM THE U-CAL DAVIS AND THEN HE DID A POST DOC WITH BEN BARRIS AT STANFORD AND IS NOW BACK THERE AFTER A DETOUR THROUGH UC SAN DIEGO. LAST YEAR, HE HAD AN INCREEING--INTRIGUING PAPER ON NEURAL ACTIVITY PROMOTES LONG DISTANCE TARGET SPECIFIC REGENERATION OF ADULT RETINAL AXONS. SO HE IS WORKING IN THE TOPIC OF HOW TO CONNECT THE RETINAL GANG LIAISON AN CELLS INTO CENTRAL PROCESSING WHERE THE SIGNALS CAN SUBSERVE VISION. HE HAS HAD OR HAS AGI FUNDING. HE ALSO HAS A NUMBER OF SUBSTANTIAL AWARDS IN HIS BACKGROUND, THE Mc KNIGHT NEUROSCIENCE AWARD AND THE COGAN AWARD FOR CONTRIBUTIONS TO VISION SCIENCE. ANDY THANK YOU FOR TAKING TIME TO COME. WE COOLED OFF THE TEMPERATURES SPECIAL FOR YOU SO THAT YOU COULD BE GLAD TO GET BACK TO STANFORD. THE TITLE OF ANDY'S TALK IS VISUAL RESTORATION BRIDGES AND GAPS TO CURING BLINDNESS IN HUMAN. [ APPLAUSE ] >> FIRST OF ALL, I'M DELIGHTED TO BE HERE. I'VE BEEN HERE A COUPLE TIMES AND EACH TIME I'M STRUCK BY HOW LARGE THIS PLACE IS AND HOW MUCH INCREDIBLE SCIENCE IS HAPPENING AND MEETING WITH THE POST DOCS AND STUDENTS IS ALWAYS A HIGHLIGHT AND GRADUATE PROGRAMS ARE NOT TYPICALLY ASSOCIATED WITH THIS, BUT AFTER LUNCH WITH THEM, THE FUTURE IS VERY BRIGHT AND IT EXCITES ME. I'VE BEEN WORKING ON VISION SCIENCE FOR ONE FORM OR ANOTHER FOR 18 YEARS SO I FELL INTO THIS GANG LIAISON AN CELL THING PRETTY YOUNG. I'M A LIFER, I WILL STAY IN THIS GAME. I THINK THERE'S A NUMBER OF IMPORTANT PROBLEMS THAT BASIC SCIENTISTS AND CLINICIANS BOTH NEED TO WORK ON AND I'M VERY HOPEFUL AND GRATEFUL FOR THE AGI, NOT JUST BECAUSE WE RECEIVED FUNDING FOR THEM BUT WONDERFUL TO HAVE SOME POINT AND DIRECTION IN WHERE TO APPLY FOR FUNDS AND SOME OF THE REALLY CRITICAL--THE FACT THAT PEOPLE STOOD BACK AND THOUGHT VERY CAREFULLY ABOUT THE PUBLIC, AND PEOPLE HERE AT NIH AND NEI AND THOUGHT CAREFULLY ABOUT WHAT WERE THE KEY PROBLEMS TO ATTACK, SO MANY GREAT PROBLEMS TO ATTACK AND SO I THINK REAL PROGRESS IS BEING MADE AND I'M DELIGHTED TO BE PART OF THAT PROGRESS AND LOOK FORWARD TO MORE. TODAY, AND I WANT TO THANK THE DOCTOR FOR THE GENERATION INTRODUCTION AND TEEF BECKER FOR BRINGING ME OUT TO TALK ABOUT OUR WORK SO FAR. SO TODAY I WILL TALK ABOUT VISUAL RESTORATION. I'M NOT AN M. D. NOT A CLINICIAN BUT I DECIDED WHOLE HEARTLY A FEW YEARS OKAY AT SAN DIEGO WORKING IN THE LAB TO RELATE TO ISSUES RELATED TO GLAUCOMA AND VISUAL RESTORATION AND WE TALK ABOUT WHY THAT IS LATER PERHAPS BUT I ALSO HOPE TO TALK A BIT ABOUT SOME OF THE WORK WE ARE DOING ON LINKING EMOTIONS TO VISION AT THE END IF THERE'S TIME BECAUSE I KNOW THERE'S FOLKS FROM THE EYE INSTITUTE AND THAT'S BOTH BECAUSE I'M EXCITED ABOUT THAT WORK BUT BECAUSE I THINK IT DOVETAILS NICELY WITH THE TECHNOLOGIES FOR ADDRESSING BLINDNESS. SO MY LABORATORY WORKS ON THREE ASPECTS OF VISION, IT'S A PRETTY LARGE GROUP OF 15 PEOPLE, A THIRD WORK ON VISUAL CIRCUIT ARCHITECT AND YOU ARE ASSEMBLY. THE GENES AND MOLECULES THAT SET UP THE VISUAL SYSTEM SO THAT ONE CAN SEE. A GOOD PORTION OF MY LAB, MORE THAN A THIRD IS WORKING ON VISUAL REPAIR, IN PARTICULAR, WHY GANG LIAISON AN CELLS DEGENERATE IN GLAUCOMA, A PROCESS THAT'S POORLY UNDERSTOOD, AS WELL AS TRY TO DEVELOP THERAPEUTICS AND STRATEGIES FOR THERAPEUTICS TO RECONNECT THE EYE TO THE BRAIN WHEN THE GANG LIAISON AN CELLS ARE INJURED OR DAMAGED BY GLAUCOMA. AND THEN A NEW ASPECTS OF MY LAB THEN THAT I'M EXCITED ABOUT, I'VE ALWAYS WANTED TO WORK ON THIS PROBLEM, WHY IS IT, THAT CERTAIN SENSATIONS ARE MERGED TO CERTAIN EMOTIONS AND HOW DOES THAT OCCUR IN THE BRAIN. IT'S RAPID. IT'S IMPORTANT. IT'S A BIG PART OF OUR DAILY LIFE AND WHEN IT GOES, WHEN THOSE SYSTEMS GO AWRY, YOU GET THINGS LIKE PTSD, CHRONIC ANXIETY AND TRAUMA. SO IT'S IN THE VISUAL SYSTEM SENSE, IT'S AN INTERESTING PROBLEM, SO I HOPE TO GET TO A BIT OF THAT AT THE END. SO A LITTLE BIT OF BACKGROUND THAT SETS UP WHERE WE'RE GOING, AS MANY OF YOU KNOW, NEURONS IN THE ADULT MAMMALIAN CNS FAIL TO REGENERATE AND CELLS IN THE RETINA, GANG LIAISON AN CELLS IRCLUEDED ARE LONG PROJECTING EXCITEATORY NEURONS AND THEY DON'T REGENERATE AFTER INJURY, AT LEAST NOT IN MAMMALS, IN COLD BLOODED VERTEBRATES THEY REGENERATE AND THAT'S AN INTERESTING PROBLEM. BUT AS A RESULT OF THIS CNS FAILURE, AS IT'S OFTEN CALLED, NEURODEGENERATIVE DISEASES IN WHICH SPINAL OR BRAIN ATROPHY OR INJURY TO THE SPINE OR CONGENITAL OR DAMAGE INDUCED OR DISEASE INDUCED CONDITIONS THAT LEAD TO BLINDNESS, GENERALLY ARE IRREVERSIBLE AND THE COMMON TREATMENTS INVOLVENINGS LIKE WHEELCHAIRS, A LOT OF OUTSOURCING OF SUPPORT, SEEING EYE DOGS, CANES AND SO FORTH BUT THE TECHNOLOGY FOR REPAIRING THE SYSTEM AND GETTING BLIND PEOPLE TO SEE AGAIN IS REALLY THE GOAL AND CURRENTLY THERE ARE NO SUCH TECHNOLOGIES. WHY DOESN'T THE CNS REGENERATE? SO I'M GIVING A LOT OF INFORMATION IN A BRIEF SLIDE BUT THERE ARE EXTRINSIC FACTORS, THAT IS THERE ARE THINGS IN THE NEURONS, GLIAL SCARS REPELLANT CUES AND CHEMICAL CUES THAT ACT AS BARRIERS BOTH PHYSICAL AND NEUROCHEMICAL TO PREVENT NEURONS FROM REGENERATING IT'S RATHER CURSORY BUT FOR THE SAKE OF TIME IT ILLUSTRATES THE PRINCIPLE. THERE ARE FACTORS AS THEY AGE, THEY GROW MUCH MORE SLOWLY THAN THEY DID DURING DEVELOPMENT, AXONS, THIS PRINCIPLE WAS DISCOVERED IN RETINAL CELLS BY JEFF GOLD BERG WHEN WAS A GRATDUATE STUDENT IN BEN VALID AND RELIABLEIS' LAB, THEY GROW VERY FAST BUT BY THE TIME THEY REACH ADULTHOOD, THEY DON'T REGENERATE OR GROW VERY MUCH AT ALL. THEY MIGHT EXTEND A COUPLE MICRONS AND THAT IN ITSELF IS SOMETHING THAT'S CHANGED INTRINSICALLY TO THOSE NEURONS AND THIS ISN'T RELATED TO REGENERATION BUT OF COURSE AXOT ME LEADS TO DEATH AND IF THERE'S NOTHING THERE, THERE'S NOTHING LEFT TO REGENERATE. IT'S SOMETHING THAT'S SURPRISING LITTLE ATTENTION HAS BEEN FOCUSED ON COMBINING NEUROPROTECTION WITH THE STRATEGIES IN THE SAME EXPERIMENT AND JUST TO EDITORIALIZE, I THINK THERE'S SOMETHING THAT WE NEED MORE EFFORT ON AND THERE ARE PEOPLE PURSUE THAG AND THAT'S GREAT. SO THE GOAL OF MY LABORATORY WITH RESPECT TO THE VISUAL SYSTEM AND THE AGI ARE TO DISCOVER STRATEGIES TO TRIGGER LONG-DISTANCE CNS AXON REGENERATION. RECONNECT CNS AXONS, THE OUTPUT NEURON TO THE HIGH WITH THEIR TARGETS. WE WANT TO PROBE THE SPEC ISOTOPITY OF REGENERATION, IT'S BEEN A LONG STANDING QUESTION, IF A RETINAL GANG LIAISON AN CELL, THESE SO CRITICALLY LINK IN THE EYE TO PERCEPTION AND MOTOR FUNCTIONS AS THEY OCCUR IN THE BRAINING, WHAT WE THINK OF AS SITE, IF REGENERATION OCCURS WILL THAT OCCUR SPECIFICALLY OR WILL THE WRONG--THE NEURONS CONNECT TO THE WRONG TARGETS, AND YOU CAN IMAGINE HOW DETRIMENTAL THAT CAN BE IF THE SO CALLED DIRECTIVE GANG LIAISON AN CELLS THAT RESPOND TO UPWARD MOTION, RECONNECT TO THE SIR CADIAN CLOCK IN THE BRAIN SO THAT SOMETHING PLUFS UP IN THE FIELD THE ANIMAL OR HUMAN BEING RATHER GETS RESET IN TERMINGS OF A SIR CADION CLOCK, THATIA A PROBLEM. SO MAYBE A REASON WHY THEY DON'T REGENERATE IS THAT IT'S BET TORE HAVE NO REGENERATION THAN INCORRECT REGENERATION AND THAT'S AN OPEN QUESTION AND I'LL TALK MORE ABOUT THAT TODAY. AND THEN THERE'S NOW GROWING EVIDENCE THAT THERE CAN BE FUNCTIONAL RESTORATION OF THE CIRCUITS BUT WE STILL AS A FIELD UNDERSTAND HOW MUCH REGENERATION YOU NEED. HOW MUCH DOES IT TAKE TO REGENERATE A FUNCTION LIKE SIGHT? AND HOW CAN WE INCREASE THOSE NUMBERS IF THEY NEED TO BE INCREASED AT ALL? SO THREEZ ARE CRITICAL ISSUES THAT CAME UP DURING THE INTERESTING AND STIMULATING DISCUSSIONS AND THERE'S SORT OF A CALL TO REMEMBERRAS IF YOU WILL AND WE RESPOND TO THE CALL TO ARMS. SO, THE INJURED EYE TO BRAIN PATHWAY IN MOUSE IS WHAT I WILL BE TALKING ABOUT TODAY. HERE ARE THE AXONS IN THE OPTIC NERVE ARE LABELED IN A DARK COLOR THIS, IS HOW I WILL SHOW THEM TODAY, THE GANG LIAISON AN CELLS OF COURSE IN LIVE IN THE BACK OF THE EYE AND THE NEURAL RETINA AND THESE ARE EXTREMELY IMPORTANT NEURONS WITHOUT WHICH THERE IS NO VISION BECAUSE THE EYE CAN BE FINE, THE BRAIN CAN BE FINE BUT IF THOSE WIRES AREN'T CONNECTED AND CONNECTED IN THE PROPER WAYS THERE IS NO VISION. SO, THE MODEL IS THE OPTIC NERVE CRUSH MODEL. THIS IS A COMMON MODEL IN WHICH FORCEPS IS LOWERED BEHIND THE EYE, THE RETINA IS CRUSHED AND THOSE THEY'RE LABELED WITH IA DIE AND YOU FIEBD UNDER NORMAL CONDITIONS THERE'S NO REGENERATION, NO AXONS EXTEND PASSED LEGION SITE SHOWN BY THE ASTERISK. THE QUESTION IS HOW DO WE GET THESE TO REGENERATE BACK IN THE BRAIN AND IF IT'S SUCCESSFUL HOW DO WE STUDY OR DO WE TARGET THE CORRECT LOCATIONS IN BRAIN. SO RETINAL GANG LIAISON AN CELLS DON'T REGENERATE NORMALLY AFTER OPTIC NERVE LESIONS. SO A VERY KEY DISCOVERY WAS MADE BY A LAB AT CHILDREN'S HOSPITAL HARVARD IN 2008 BY A POST DOC KEVIN PARK IN HIS OWN LAB IN MIAMI, IN WHICH THEY SHOWED THAT A GENE, MTORIS EXPRESSED AT HIGH LEVELS AND GANGLION CELLS IS SUPPRESSED BY THE P10 GENE, GOES DOWN ACROSS DEVELOPMENT AND WHAT THEY DID WAS KEVIN CREATED A SITUATION IN THE MOUSE IN WHICH HE DID A CONDITION OF THE INHIBITORS, INCREASING MTOR IN THE EYE, SORT OF MIMICKING THE LEVELS OF MTORDURING THESE CELLS, EFFECTIVELY TURNING ADULT RETINAL GANG LIAISON AN CELLS INTO THE BABY RETINAL GANG LIAISON AN CELLS WITH THE EXPRESSION LEVELS. HE DID THAT BEFORE HE CRUSHED THE OPTIC NERVE, THEN HE CRUSHED THE OPTIC NERVE, AND WHAT SHE WAS IMPRESSIVE AND I DON'T THINK ANYONE HAD SEEN THIS MUCH REGENERATION BEFORE EXCEPT IN CONDITIONS LIKE CNTF APPLICATION. IT WAS STRIKING WHAT WE SAW. AND WHAT WE'VE REPLICATED MANY TIMES IT WAS AN IMPORTANT POINT BECAUSE REGENERATION FIELD OF SPINAL CORD AND OPTIC NERVES, SORT OF A NUMBER OF FAILURES TO REPLICATE OVER THE YEARS AND IT'S A READILY REPRODUCIBLE EFFECT. THE LESION IS HERE AND MANY OF THOSE THAT STEP DOWN NEAR THE OPTIC BRAIN OVER HERE, SO THE PATHWAY TURNS OUT TO BE CRITICAL AND IT'S KIND OF CONTROVERSIAL BECAUSE YOU WORRY ABOUT CANCERS AND THINGS ASSOCIATE WIDE MTOR, BUT NONE THE LITTLE, THIS EMERGED THIS MTORMODEL HAS EMERGED AS THE CRITICAL OR MOST ROBUST MODEL FOR GENERATING LONG DISTANCE AXON REGENERATION WITH A SINGLE TREATMENT. HOWEVER, P10 DELETION ALONE INCREASING MTOR WAS INSUFFICIENT TO PROMOTE THE REGENERATION, THE AXONS MADE IT ABOUT AS FAR AS THE DISTAL NERVE AND THIS DID NOT GROW THROUGH THE KHIHIGH ASISM THROUGH BRAIN AND THEY WERE FUNCTIONAL OR VISUAL RECOVERY IN THESE ANIMALS. NOW WE SAT BACK AND THOUGHT ABOUT THIS AND WE DECIDED THAT AT SOME LEVEL, WE WANTED TO UNDERSTAND WHETHER OR NOT THE PRINCIPLES THAT WIRE UP GANG LIAISON AN CELLS DURING DEVELOPMENT WOULD APPLY TO TRYING TO REGENERATE THOSE GANGLION CELLS IN ADULTHOOD, WE KNOW DURING DISWEPMENT THAT THESE ARE ACHIEVED BY GROWING OUT INTO THE BRAIN AND OOH DENTIFYING WHICH TARGETS TO CONNECT TOO IN SPECIFIC WAYS AND THEY GROW MORE AT THAT AGE AND LATER VERY, VERY LONG DISTANCES AND THEY DO THIS IN VERY PRECISE WAYS. SO WE STARTED THINKING, MAYBE THESE PRINCIPLES THAT APPLY EARLY IN LIFE WOULD APPLY LATER IN LIFE ONCE GANG LIAISON AN CELLS ARE REGENERATING, SO WHAT ALBERT LYNN, THEN A GRADUATE STUDENT OF MINE DID WHO IS NOW AT GENENTECH, AND HE PUT GANG LIAISON AN CELLS INTO A DISH, GAVE THEM GROWTH FACTORS AND HE STIMULATED THE CELLS ELECTRICICALLY AND THE OTHER--AND OTHER GANG LIAISON AN CELLS HE DIDN'T STIMULATE. THIS SCHEMEATIZES WHAT IS SHOWN IN THE PAPER, AND THIS SHOWS THAT THE ELECTRICALLY STIMULATED CELLS WERE GROWN THAT RESEARCH MUCH LONGER AND LARGER AND THESE WERE NOT ELECTRICALLY ACCOUNTAIVE AND THEY USE PATTERNS OF ACTIVITY TO MIMIC THE SORTS OF ENDOGENOUS PATTERNS AND THAT OTHER VS STUDIED AND DISCOVERED AND STUDIED OVER THE YEARS AND THAT PROVE IMPORTANT FOR VARIOUS ASPECTS OF GANG LIAISON AN CELL WIRING AND DEVELOPMENT SO ALBERT DECIDED TO ASK THE STRAIGHT FORWARD QUESTION, DOES NEURAL ACTIVITY CONTRIBUTE TO REGENERATION OF RGC AXONS INVIVO AND THAT'S WHY MAYBE THEY DON'T GROW VERY FAR, IF WE WERE TO INCREASE THE LEVEL OF ACTIVITY, THEY WOULD GROW FURTHER. SO SINCE THESE DATA ARE PUBLISHED, I WILL MOVE THROUGH THEM QUICKLY AND I WILL HIT THE IMPORTANT POINTS BUT I WOULD BE HAPPY TO STOP AT ANY POINT AND ANSWER QUESTIONS. AT FIRST HE DIDN'T TAMPER WITH THE MTOR PATHWAY AT ALL. HE CRUSHED THE NERVE AND THEN HE GAVE THE MICE HIGH FREQUENCY AND STIMIEWLINGSZ, THOU THIS IS A LOT OF STIMULATION, THIS WAS ABOUT 10 HOURS A DAY. SOMETHING A PATIENT WOULD NOT WANT TO DO AND WE WILL REVISIT THAT IN A BIT 6789 OTHER GROUPS PUBLISHED THE FACT THAT, AND--YOU SEE SIMILAR LEVELS OF REGENERATION WASN'T VERY FAR AND NOT VERY MANY ACONS BUT APPLYING THE ACTIVITY REGIME DID SEEM TO INCREASE THE AMOUNT OF ACTIVITY IN THE GANG LIAISON AN CELLS AND PROMOTE GROWTH IN THE LESION SITE A LITTLE BIT. SO HERE'S QUANTIFICATION WITH THAT, IT WAS SIGNIFICANT AND NONE OF THEM MADE IT TO THE KHIHIGHASM WHICH MADE IT FROM THE CRUSH SITE. SO NOTHING TO GET TOO EXCITED ABOUT BUT IT WAS INTRIGUING. SO WHAT'S THE MECHANISM FOR THIS, SO WE WONDERED WHETHER VISUAL STIMULATION COULD PRODUCE ACTIVITY TO STIMULATE LONG DISTANCE REGENERATION AND HE FURTHER TEST THE ROLE OF ACTIVITY AND REGENERATION OF GANG LIAISON AN CELLS BY USING THE DREAD SYSTEM, CHEMICAL SYSTEM WE CAN TALK ABOUT IN DETAIL BUT FOR SAKE OF TIME. YOU INTRODUCE A SYNTHETIC RECEPTOR, THE KEY NUMBER IS FOUR OR THE HM3D, THE OTHER ANIMAL HAS HMFOUR D, PUT IN THE CELLS WITH THE INJECTION AND WITH THE APPLICATION OF THE DRUG CNO INTO ONE SET OF MICE OR THE LITTLER SET OF MICE, YOU COULD DECREASE OR INCREASE THE AX MOUNT OF GANG LIAISON AN CELL SPIKING. THE NICE THING ABOUT THIS ALLOWED US TO TEST HOW MUCH WAS INCREASED OR DECREASED FROM THESE CELLS AND PATCH RECORDINGS, HE'S TARGETING ONE OF THESE CELLS THAT EXPRESSES ONE OF THESE SYNTHETIC GENES AND HE SHOW SYSTEM HERE'S NOW THE HM40 SO IT'S ALL THE SPIKING SPONTANEOUSLY OR RESPONSE TO VISUAL STIMULI, SAME EFFECT SHOWS THAT IF YOU WASH, THE CELL IS SILENT SO IT'S A GENETIC TTX EXPERIENCE. AND THIS AND YOU CAN DO AND SUPPRESS ACTIVITY THROUGH THE DURATION OF THIS PERIOD AFTER THE OPTIC NERVE LESION, THE OPPOSITE EXPERIMENT--OH, EXCUSE ME. WHAT HAPPENS TO REGENERATION? THIS WAS INTERESTING IT TURNS AND THERE'S ACTUALLY THAN YOU WOULD SEE ORD NARRLY, YOU DON'T EVEN SEE THESE GO UP TO THE LESION SITE. MOAF OF THESE DIE WHEN YOU BLOCK ACTIVITY IN THIS WAY. SO THAT IS PRESERVING THE LIFE OF THE GANG LIAISON AN CELLS AFTER INJURY. SO HERE, PATCH CLAMP RECORDINGS FROM THREEZ 3D INFECTED CELLS SO THAT THE CELLS MERELY SPIKING ALONG, WASHING CNO, AND NOW BECAUSE THERE'S SECOND MESSENGER SYSTEM, LEADS TO OPENING OF POTASSIUM CHANNEL AND WHAT HAPPENS IS THE CNO, CAUSES A DRAMATIC INCREASE ABOUT A DOUBLING IN THE SPIKE OUTPUT OF THE GANG LIAISON AN CELL, BOTH SPONTANEOUS AND EVOKED. SO WHAT HAPPENS WHEN YOU REGENERATE IN ACTIVITY IN THIS MANNER, AND WHAT ALBERT SAW, I'LL JUST SO YOU SHE'S ARE THE LOW MAGNIFICATION VIEWS SO YOU COO SEE, HERE'S THE CHIASM, HERE'S THE AREA PROXIMAL AND YOU CAN SEE MANY ARE REGENERATING THROUGH, DEFINITELY MORE THAN WE SAW IN TERMS OF USING VISUAL STIMULATION, THESE GENETIC APPROACHES ARE INTERESTING, IT'S ONE VIRAL INJECTION AND THEN REPEATED INJECTIONS IN THE CNO OR GUT OR DRINKING WATER. A FEW AXONS MADE IT TO THE MIDDLE OF THE OPTIC NERVE, VERY FEW MADE IT IF ANY, AND THAT'S DEBRIS, THOSE ARE DEGENERATED AXONS YOU SEE THERE. SO THE MORE ACTIVITY, THE MORE LONG DISTANCE REGENERATION. IT'S STILL A SMALL NUMBER OF AXONS BUT NOW WE WERE HOOKED ON THIS AND WE THOUGHT, WOW, WHAT WOULD HAPPEN IF WE STARTED COMBINE THIS WITH OTHER TREATMENTS THAT TICKLE THE GANG LIAISON AN CELLS IN TERMS OF MOLECULAR GROWTH PROGRAMS. ANOTHER POINT OR TWO ABOUTLET PROMECHULAR GROWTH PROGRAMS, THIS IS QUANTIFICATION, THIS IS THE MOST REGENERATION WE SAW, SO, THE PATHWAY THAT WE WANT TO FOCUS ON WAS THIS MTORPATHWAY, YOU CAN INCREASE MTOR BY BLOCKING INHIBLETTOR, P10 IN PREVIOUS SLIDES, AND KEVIN PARK AND OTHERS HAVE DONE, SO WE DECIDE TO TAKE A VIRAL STRATEGY BECAUSE IN TERMS OF TRANSGENIC HUMANS THAT WILL BE A TOUGH ONE. ALTHOUGH CRISTP AND INTERESTING WITH ITS SPECIES SO WE DECIDED TO TRY TO INJECT AN ADNO VIRUS INTO THIS AND WE SHOWED THAT THE IN ADULT RETINA, THIS IS RHEBONE LEADS TO A ROBUST AMOUNT OF CELLS AND THE AMOUNT OF MTORTHAIZ THEY EXPRESS. HERE'S DOUBLING OF THE AMOUNT OF CELLS AND THE AMOUNT THAT THEY EXPRESS. OKAY, SO NOW, WHAT HAPPENS DOES THIS DELETION AND I WILL SHOW YOU WHEN YOU INCREASE RHEB ONE AND YOU SEE SOMETHING SIMILAR TO WHAT HAPPEN WHEN IS YOU DECREASE P10 AND YOU SEE A LOT OF REGENERATION AND WE KNOW THIS IS OCCURRING THROUGH ACTIVATION OF THE MTORPATHWAY BECAUSE THE EFFECT AND BLOCKED BOO THE APPLICATION WHICH BLOOKS THE MTOR PATHWAY. SO THIS CAN INCREASE REGENERATION, NOT MUCH, INCREASING TACKIVITY CAN INCREASE REGENERATION BUT NOT MUCH, BUT NONE OF THEM MAKE IT ALL THE WAY OUT TO THE CHAIASM, SO ALFRED COMBINED THE DATA FROM THE TWO. SO CAN THEY COMBINED PROMOTE GREATER REGENERATION. HE LESIONED THE OPERATING GLOBALLYIC NERVE, INFECTED THAT EYE AND THE GANG LIAISON AN CELL WITH RHEBONE AND HE FOUND THAT THE COMBINED TREATMENT DID LEAD TO AXONS GROWING DOWN THE OPTIC NERVE BUT NONE OF THEM MADE IT TO THE CHIASM, AND WE FIGURED OUT ABOUT SIX MONTHS TO A YEAR FIGURING OUT WHAT WAS GOING ON BECAUSE EVERY ONCE IN A WHILE HE WOULD SEE AN ANIMAL THAT WOULD REGENERATE IN THE BRAIN, GIVING WAY TO THE NEXT PART OF THE TALK AND WE COULD NOT FIGURE OUT WHAT WAS GOING ON. AND I SAID WALK ME THROUGH THE ENTIRE EXPERIMENT AND WE WERE DOING VISUAL TESTING ON THE ANIMALS EVEN THOUGH THEY WE CAN'T DO VISUAL TESTING AFTER YOU KILL THE ANIMAL OBVIOUSLY SO YOU HAVE TO DO BEFORE YOU SACRIFICE THE ANIMAL TO LOOK AT NERVE REGENERATION AND SOME OF THOSE TESTS, ALBERT WAS FORCED TO CLOSE THE OPPOSITE EYE, FOR REASONS RELATED TO VISUAL TESTING I'LL TALK ABOUT A BIT LATER AND IN THOSE ANIMALS WE SAW LONG DISTANCE REGENERATION. I THOUGHT WELL THIS IS REALLY WEIRD AND SOME OF YOU PROBABLY ALREADY REALIZE WHY IT'S WEIRD AND I THINK WE HAVE A HANDLE ON THIS NOW. SO ALBERT SET UP THESE CONDITIONS ALL OF THEM BLIND CONDITIONS SO HE DOESN'T KNOW WHAT ANIMALS ARE GETTING INJENTHED WITH THE RHEB -ONE VIRUS AND CONTROL VIRUSES SO HE SAW THAT IF HE LESIONS THESE NERVES, THE ONES THAT HAVE THE RHEB ONE TREATMENT, BUT THE OPPOSITE EYE WAS CLOSED TO BIAS STIMULATION THROUGH THE OPEN DAMAGED AND TREATED EYE, WHAT HE SAW WAS THE AXONS MAKE IT INTO THE CHIASM, AND ACTUALLY LATER IN THE BRAIN AS WELL. SO THE DISTAL ORPTIC NERVE AND THROUGH THE CHIASM. AND WE WERE SLIGHTLY DISTURBED BECAUSE TO OUR KNOWLEDGE THERE ARE NOT MANY CONNECTIONS IF ANY BETWEEN THE TWO EYES, AND THAT'S BEEN REPORTED AND THERE'S RETINAL-RETINAL CONNECTIONS IN BIRDS. THERE MAY BE SOME BUT THEY'RE NOT VERY MANY IN ANY CASE BUT THERE'S A SHORT WINDOW IN WHICH AXONS FROM THE TWO EYES ARE POSITIONED TO INTERACT WITHIN THEIR TARGETS AND FACTORS THAT MIGHT IMPACT THIS, AND THIS IS SOMETHING FOR DISCUSSION AT THE END. SO ALBERT WAS EXCITED BY THIS AND QUANTIFIED IT ACROSS A NUMBER OF ANIMALS AND THAT'S SHOWN HERE IN PURPLE WHICH JUST TO COLOR CODE TO MAKE IT SIMPLER WHEN HE CLOSES ONE EYE, DOES THE VISUAL STIMULATION, AND 10-12 HOURS A DAY, EXCUSE ME AND SEES A LOT OF REGENERATION ALL THE WAY TO CHIASM BUT NOT IN OTHER CONDITIONS. SO WHEN I CLOSE, OTHER EYE OPEN HAS TO BE THIS EYE, YOU CAN'T CLOSE THIS EYE SO HE DID ALL THE DERIVATIONS, THERE WERE 27 SUPPLEMENTAL FIGURES IN IN PAPER FOR ALL THE DERIVATIONS, AND PLUS, INCREASE IN ACTIVITY IN GANG LIAISON AN CELLS. NOW IN FIELD OF REGENERATION IS FILLED WITH RESULTS THAT PEOPLE AREN'T MOTIVATED OR FUNDED OR JUST DON'T WANT TO REPORT DOING AND NOT SEEING THE SAME THING. I LIKE TO SLEEP WELL AT NIGHT. I TRUSTED ALBERT AND I STILL TRUST HIM BUT YOU WANT TO SEE THIS STUFF VERIFIED INDEPENDENTLY AND WE ALSO KNOW THAT THE HM3D APPLICATION LEADS TO A ROBUST NEURAL ACTIVITY THAN DOES THE VISUAL SIMULATION AND I SHOWED YOU THAT EARLIER AND WE KNOW THAT PRACTICES PRECORDINGS AND SPIKE OUTPUTS FROM INVIVO ARE MUCH LESS THAN THESE GENETIC TOOLS SO I'M DELIGHTED TO SAY, THAT ALEX'S LAB, DOWN THE ROAD, FROM THE SCHOOL OF MEDICINE, ALEX HAS BEEN DOING BEAUTIFUL WORK, THE MOLECULES THERE AND IS EAGER TO MOVE INTO THE REGENERATION FIELD AND APPLY HIS KNOWLEDGE OF AXON GUIDANCE, LIGANDS AND MOLECULES HE DISCOVERED AND CHARACTERIZED TO THAT SYSTEM. I WILL TALK MORE ABOUT THIS BUT ALEX'S GROUP IS NOW INDEPENDENTLY VERIFIED THIS RESULT THAT THE COMBINATION OF HM3D WITH THE INCREASE IN THE RHEB ONE, LEADS TO REGENERATION, THE AXONS DOWN THE OPTIC NERVE AND THIS IS POST CHIASM, THIS IS WHERE THE IT'S HEADING INTO THE BREAK AND THE OPTIC TRACKS SO THAT MAKES ME HAPPY AND RELIEVED. YOU KNOW REPPLICATION IS A TRICKY THING BECAUSE THEN PEOPLE THINK ABOUT DRVESES IN MOUSE STRAINS AND HOUSING CONDITIONS AND THEN THERE'S VISUAL INVOLVED AND LIGHT-DARK CYCLES SO THERE'S CERTAINLY MORE WORK TO BE DONE AND I THINK REPLICATION, ESPECIALENTIALLY IN THE FIELD OF REGENERATION WILL BE VERY IMPORTANT AND AGAIN, I'M DELIGHTED THAT AGI HAS HIGHLIGHTED THIS AS ONE OF THE CRITICAL BULLET POINTS. REPLICATION OF RESULTS THAT EVALUATE PROBUSTNESS. I THINK EVERYONE IN THE FIELD IS VERY WELL INTENTIONED BUT YOU KNOW ANY NUMBER OF THINGS CAN HAPPEN IN TERM OF MOUSE STRAIN SAYS AND-OF TRAINS AND THIS IS GREAT. SO WHERE DO THEY GO? DO THEY GO TO THE RIGHT PLACE? WRONG PLACE? I WILL GIVE YOU THE MESSAGE QUICKLY. THIS SAY SCHEMATIC THIS, IS THE EYE, THE VISUAL TARGETS IN THE BRAIN AND A NORMAL MOUSE, SOME OF THEM ARE DEVOIT OF BLACK LABEL HERE IS THEY SET A MEDIAL TO THE PANEL OF THE SCREEN BUT NONETHELESS THERE ARE 40 DIFFERENT TARGETS AND SO YOU LOVE TO KNOW, DO THE CELLS GO TO VISUAL TARGETS OR WANDER INTO THE AUDITORY TARGETS SO WE KNOW THAT VISUAL NEURONS IN PARTICULAR, GANG RYAN CELLS ARE PERFECTLY HAPPY TO GROW INTO THE THALAMUS IF THEY'RE ALLOWED WHERE THEY DRIVE THE AUDITORY PERCEPTIONS. WE DON'T WANT THAT, RIGHT? SO THE GOOD NEWS IS THEY STAY IN THE VISUAL SYSTEM, SO HERE'S A SAGEITAL VIEW OF MICE THAT RESEECHED, SCHEMATIC OF THESE MICE THAT STIMULATE LONG DISTANCE REGENERATION AND ALBERT AWE AN AXON OR TWO IN THE SUPER CHIASMATIC NUCLEUS, HE DIDN'T DISCOVER IT FIRST, HE SPENT A LOT OF TIME THERE, CONGRATULATIONS HE'S HERE. HE HAS DONE WONDERFUL WORK. VLGN, ROLE IN EYE MOVEMENTS AND THE FAMOUS DLG AND THE MOST FAMOUS SUBCORTICAL TARGET AND IT RELAYS INFORMATION UP TO VISUAL CORTEX. THEY'RE THE PROFILES WERE LOOKING KIND OF DEGENERATED AND THESE ARE NOT FABULOUS TERMINALS BUT WE SAW TERMINALS THERE. I WILL REVISIT THAT POINT ABOUT DEGENERATION OF REGENERATED AXONS IN A BIT. THE NUCLEUS, WHICH IS INVOLVED IN PUPIL REFLEXES WE SAW AXONS THERE. AND THEN WITH BRAIN STEM--BRAIN STEM TARGETS THEY'RE INVOLVED AND YOU MOVE OUR HEAD OR EYES THE VISUAL WORLD SLIPS ON THE RETINA BUT UNLIKE A PROORLY STABILIZED IMAGEOT CELL PHONE, WHICH IS BLURRY YOU GENERATE A EYE MOVEMENT AND MOTHER NATURE SOLVED THAT PROBLEM FOR US AND IN THE MOST DISTAL TARGET THE SUPERIOR, WHICH IS INVOLVED IN DREBLGHTED EYE MOVEMENTS IN THE LOCATIONS IN THE VISUAL FIELD AND YOU HAVE MANY PEOPLE HERE WHO HAVE DONE BEAUTIFUL WORK ON THAT, INCLUDING BOB WURTZ, OF COURSE. THERE'S REGENERATION, I WANT TO SPEND A BIT OF TIME WITH THIS SLIDE SO THERE'S IMPORTANT THINGS, SO THERE'S NO MISCOMMUNICATION. FIRST OF ALL, IT'S FEW AXONS PROBABLY LESS THAN A HUNDRED. THERE ARE 10,000 GANG LIAISON AN CELLS IN THE MODEL CITIZEN US AND HERE WE EVALUATED EVERY TARGET 10,000 PER EYE AND COULD BE MANY MORE BUT ABOUT 10,000, PROBABLY ABOUT A HUNDRED GANG LIAISON AN CELLS. THEY SPREAD THESE OUT BROADLY BUT WITH THE PLASTICITY AND THE POST SYNAPTIC STRUCTURE RESCUE FUNCTION. THE QUANTIFICATION OF THIS, I HIGHLIGHTED SO YOU DON'T HAVE TO READ THIS WHOLE TABLE, BUT ZERO OUT OF 16 OR 13 MICE, NOT REGENERATE INTO THE TARGET SO THE MOST ROOF THEERAL OF THE CHIASM, ARE HERE, AND THIS IS THE SUPERIOR CO LICKULOUS ARE HERE. FIVE OUT OF 10, SIX OOF THE OF 10, SO FORTH, WHAT IS THIS? THESE ARE ANIMALS THAT HAD ONE EYE CLOSED, VISUAL STIMULATION AND THE RHEB-ONE, MTOR, MTOR,--ACTIVATION, AND SOMETHING WHERE THE EYE WAS REMOVED, THE MOST STRONGLY AND REMOVES THE PHYSICAL SUBSTRATE OF POSSIBILITY OF OTHER INTACT AXONS IN THE BRAIN BLOCKING REGENERATION SO THAT WAS THE RATIONAL FOR DOING THIS. YOU NEED THESE THREE THINGS IN ORDER TO GET LONG DISTANCE REGENERATION, THE GREAT NEWS IS THE AXONS STAYED IN THE VISUAL SYSTEM, NONE OF THEM WANDERED INTO THE AUDITORY SYSTEM BUT IT TURNS OUT THAT'S THE CASE SO SOMETHING IS INHIBITORS IN THOSE MODALITIES OR SOMETHING IS KEEPING THISEM IN THE CORRIDOR OF THE VISUAL SYSTEM, EITHER STRUCTURAL OR CHEMICAL. A COUPLE QUICK CONTROLS. YOU KNOW THIS IS IN THE MANUSCRIPT BUT WE DID DO CONTROLS TO MAKE SURE WE WERE NOT LOOKING AT SPARED AXONS, HOW DO YOU KNOW IF YOU ACTUALLY LESION THE AXONS WELL ENOUGH. IT'S VERY TOUGH. NOT LIKE OUR STUDIES OF GLAUCOMA AND DAVID CULKINS AND SAPPING TONS YOU CAN SEE THE MODEL OF GLAUCOMA, YOU CAN MEASURE PRESSURES EVERY DAY AND YOU CAN KNOW, THIS MOUSE HAD AN INCREASE OF 30-GRAMS OF MERCURY AND SO YOU CAN KIND OF GAUGE THE SIZE OF THE EFFECT, THE MAGNITUDE OF THE EFFECT AND THE MAGNITUDE OF THE MEASURE CHANGE AND YOU CAPTAIN DO THAT IN REAGAINERATION AND YOU ARE GUESSING WHAT HAPPENED AFTER THE EXPERIMENT IS OVER. SO ALBERT DIDN'T AN EXPERIMENT WHERE HE PRELABELLED ALL THE CELLS AFTER IT DIED AND THEN AFTER THAT HE RELABELED THEM IN THE SAME EYE WITH THE GREEN DYE AND NONE OF THE GREEN STUFF IS ALSO MAGENTA AND NONE OF THEACONS WERE SPARED AXONS--EXCUSE ME--NONE OF THEM WERE SPARED AND YOU SEE THE DIE BACK OF THE PRELABELLED ONES. SO THIS CONFIRMED IT THERE WASN'T SPARING OF AXONS IN THESE ANIMALS. HE ALSO DID AN EXPERIMENT WHERE HE SAID, WELL IF IT'S SPARING OF AXONS THEN THEY SHOULD BE VISIBLE IN THE BRAIN AFTER THE PROTOCOL IS INITIATED BUT IF THERE'S A KIND OF SUCCESS OF ACCUMULATION OF GROWING NEURONS, KIND OF A FLEET OF NEURONS IN THE BRAIN AND IF YOU SACRIFICE THEM AFTER ONE WEEK, TWO WEEK OR THREE WEEKS THEN HE OUGHT TO SEE LESS MORE AND MORE REGENERATION AS TIME WENT ON AND THAT'S WHAT HE SAW. SO AS TIME WENT ON HE SAW MORE AND MORE REGENERATION INTO THE BRAIN. SO COUPLE WEEKS AFTER VERSES ONE WEEK AFTER, SO IT'S REALLY A THREE WEEK SYSTEM WHERE THE AXONS HIT THE BRAIN AND THAT'S WHEN THEY GROW REALLY FAST AND THATY AN ENTREESING THING WE EVAPORATE FOLLOWED UP BUT I HOPE SOMEONE DOES, THESE GROW FASTER IN THE BRAIN THAN THEY DO IN THE OPTIC NERVE. NOW THIS IS A QUESTION ABOUT FES FISCHERITY. SO ONE THING WE WERE REALLY INTERESTED IN IS WHETHER OR NOT THESE GANG LIAISON AN CELLS OF WHICH THERE ARE ABOUT 30 DIFFERENT TYPES AND I THINK THE NUMBER IS--I'VE BEEN INVOLVED IN THIS EXPOSURE TO RADIATION TO TYPE GANG LIAISON AN CELLS. I THINK WE ALL AGREE NOW BASED IN LARGE PART DUE TO THE BEAUTIFUL YORK OF TIME OILER AND LITTLERS IN--OTHERS IN THIS EFFORT, EACH ENCODING A DISTINCT QUALITY OF INFORMATION AND SIGNALING IS THAT TO THE BRAIN AND EACH ONE OF THOSE CELL TYPES PROJECTING TO A UNIQUE COMBINATION OF ABOUT FOUR OR FIVE OF THE POSSIBLE 40 DIFFERENT TARGETS IN THE BRAIN. SO IT'S A PRETTY COMPLEX PUZZLE. SO, WE'VE DEVELOPED THESE DIFFERENT MOUSE LINES OR CHARACTERIZE DIFFERENT MOUSE LINES AND EACH IS A SET OF CELLS THAT CONNECTS TO THE TARGETS IN THE BRAIN, NORMALLY IS LABEL WIDE A FLUORESCENT PROTEIN SO I TRIED TO MAKE THIS AS SIMPLE AS POSSIBLE, SO ALL THESE RESULTS ARE PUBLISHED AND THE MICE ARE FREELYY AVAILABLE THANKS TO THE GEN-SAT PROJECT. THESE IS THE CELLS HERE AND HERE BUT NOT TO THE OTHER TARGETS, BLUE CELLS PROJECT HERE, HERE, AND HERE, BUT NOT OTHERS AND SO FORTH. SO ALBERT DID THESE EXPERIMENTS AND ASKED WHETHER OR NOT THE INDIVIDUAL GANG LIAISON AN CELLS IF THEY'RE PART OF THE REGENERATING COHORT WHETHER OR NOT THEY WIRE UP TO THE CORRECT TARGETS AND THE RESULTS OF THOSE--HERE'S ANOTHER MOUSE LINE. WE HAVEN'T HIT SATURATION, WE DON'T HAVE MARKERS FOR ALL 30 BUT COLLECTIVELY AS A FIELD WE MARKERS FOR 20 WHICH IS PRETTY EXCITING. SO AT LEAST FOR THE ALPHA-LIKE GANG LIAISON AN CELLS, THESE ARE SIMILAR TO THE PARASOL GANG LIAISON AN CELLS IN PRIMATE, THESE AXONS REGENERATE BACK TO THE KREBLGHT TARGET SO THIS IS PANELS ALL GANG LIAISON AN CELLS COLLECTIVELY WITH THE CTB DYE, AND PSEUDOCOLLORRED HERE IN BLACK AND THEN THEY'RE MERGED AND IT SHOWS THAT THESE GREEN AXONS AND WE SAW THAT THE GREEN AXONS ARE PRESENT IN THE NORMAL TARGETS OF THE CELLS SO FOR INSTANCE THESE CELLS HERE AND THE DLGN AND OPSC, AND THE FUNCTIONS OF THESE ARE DIFFERENT AND THE NAMES, FULL NAMES ARE NOT IMPORTANT FOR NOW, BUT WHAT ALBERT SAW IS THAT THE GREEN AXONS WERE PRESENT WITHIN THOSE SPECIFIC TARGETS AND THAT MEANS THAT THESE AXONS ARE GROWING BACK DOWN THE NERVE THROUGH THE CHIASM, IN THE BRAIN AND FINDING AND RECONNECTING TO THE CORRECT TARGETS. HORMONE'S ONE AXON, IT'S EMBEDDED IN AN OCEAN OF AXONS. WHY ARE SOME GREEN AND NOT MAGENTA, AND THAT'S BECAUSE THEY'RE FROM THE OPPOSITE EYE AND ALBERT INCLUDED DAILY BASIS THEA AND I INVITE YOU TO LOOK AT THOSE IF YOU LIKE, IN WHICH HE THEN INCLUDED OPPOSITE EYE AND ALL OF THOSE ARE LABELED IN THE BRAIN AND THOSE HAD TO BE AXONS THAT WERE REGENERATED AND LABELED AFTER THE REGENERATION, SO GANG LIAISON AN CELLS KNOW WHERE TO GO AND WHERE NOT TO GO AND I WILL MENTION THAT THERE'S THREE OR FOUR SUPPLEMENTAL FIGURES THAT ILLUSTRATE NONE OF THE TARGETS, I WILL NOT GO THROUGH THEM FOR SAKE OF TIME. BUT NONE NONE THE OF THE 40 TARGETS THAT DON'T NORMAL RECEIVE THOSE F ALPHWACELLS GET INPUT IN THE REGENRATIVE CONDITION AND WHEN HE LOOKED AT THESE PHOTOSENSITIVE AKA MEL OPERATING GLOBALLYS, THEY REGENERATE AND THE ONE THATION REGENERATE GO TO THE RIGHT PLACE AND NEVER TO THE WRONG PLACE, I THOUGHT, WELL THIS IS FANTASTIC, GANG LIAISON AN CELLS KNOW THEIR WAY HOME, AND THEY'RE REACTIVATING DEVELOPMENTAL PROGRAM AND I'LL REVISIT THAT IN A FEW MINUTES WITH SOME CHEMICAL OR MOLECULAR DATA AND WE NOW HAVE A NICE SUITE OF VISUAL BEHAVIORS MANY OF WHICH TAP INTO THE TARGETS IN KNOWN WAYS LIKE THE OPTICAL IMAGES O KINETIC REFLEX AND THE PUPILARY REFLEX TO BLUE LIGHT AND CLIFF WHICH IT RELIES ON CORTEX THROUGH THE GENICULET, AND IT THE MOUSE STEPS OFF THE PLATFORM FROM DEEP TO SHALLOW, BUT I MICE DON'T KNOW THAT. AND THE LOOMING RESPONSE IN RELATION TO THE FEAR WORK IN WHICH AN OVERHEAD DARK EXPANDING STISK IS SHOWN, AND THE MOUSE FREEZES OR FLEES TO SAFETY. SO HE EVALUATE VISUAL DEHAIEVERS IN THESE MICE TO SEE IF THE AXONS REGENERATE BACK INTO THE BRAIN. VERY FEW AXONS. SO HE DID THIS, AND I WILL SUMMARIZE HERE, PUT SIMPLY, THE OPTICAL IMAGES O KINETIC REFLEX, THIS IS A NONLESION ANIMALS HOOEY DID IT IN THIS A WAY IN WHICH YOU A NUMBER OF TRIALS THE MICE TRACK THE VISUAL STIMULUS, THERE ARE A NUMBER OF WAYS TO DO THAT BUT THAT'S WHY IT'S 30% OF TRIALS. OCCASIONALLY THE MICE DON'T PAY ATTENTION, THEY DON'T LIKE DOING PSYCHOPHYSICS EITHER, UNLESS THEY'RE HAD-FIXED AND THEN THEY WILL FORCED TOO AND WILL DO IT MORE OFTEN. THE LESION WAS EXCITING, IT WAS ATTRACTED. THERE WAS NOTHING IN AN LESION OF AN UNTREATED ANIMAL. TWHA WAS FSN'T SURPRISING. IN THE PUPIL REFLEX PALGT WAY, WE SAW AXONS GRG TO THE TARGETS OF THE REFLEX BUT WE DIDN'T SEE ANY RECOVER OF FUNCTION AND THE AXONS LOOK HEALTH TOW US AND THIS POINTS TO THE FACT THAT STRUCTURE AND FUNCTION CAN SOMEHOW--THEY DEFINITELY RELATE BUT IT CAN BE MISLEADING WHEN YOU LOOK AT STRUCTURE. THE VISUAL CLIP SHOWED NO RECOVER EXPE THAT WAS DEPRESSING BECAUSE WE SAW HIM BACK INTO THE RGC, AND THEY WOULD LIKE THE SIR CADIAN CLOCK TO THE LIGHT AND DARK CYCLE, THAT'S IMPORTANT PHYSIOLOGICAL MANIFESTATION, AND WHAT WE WANT IS THE MANIFESTATION OF SITE BUT THEY WERE BLEVY, AND THEY WERE NOT HEALTHY OR ABLE TO MEADIAT SIN APTIC TRANSMISSION OR POST SIN APTIC CELLS. SO VISION ISN'T COMPLETELY RESTORED BUT THERE'S PAICIAL RECOVERY OF FUNCTION IN THESE TWO BEHAVIORS, THE OPTICAL IMAGES O KINETIC REFLEX AND THE LOOMING RESPONSE. A LOT MORE WORK NEEDS TO BE DONE THERE. SO TO SUMMARIZE THE PORTION OF THE TALK, I TOLD THAT YOU NEURONAL ACTIVATION CAN BE USED AS A STIMULUS BUT ALONE IN COMBINATION WITH OTHER INFO ABOUT THE BRAIN, IT CAN HELP GET THOSE TO GET GANG LIAISON AN CELLS TO GET THOSE BACK IN THE BRAIN ESPECIALLY WITH YOU BIAS TOWARD THE INJURED EYE? THIS MIGHT NOT BE THAT SURPRISING RIGHT? MY GREAT GRAND PARENTS, [INDISCERNIBLE], OBVIOUSLY, MOST FAMOUS FOR THEIR WORK ON DISCOVERING THE SPHRUCTURE FUNCTIONAL RERELATIONSHIPS IN THE VISUAL SYSTEM AND OCULAR DOMINANCE PLASTICITY SAID THIS MUST BE THE CASE, I HAVEN'T SHOWN THEM THE DATA BUT I MAY SEE THEM AT THE PEW MEETING, BUT DAVID PASSED AWAY RECENTLY, SADLY, THEY WOULD SAY, THERE'S COMPETITION BUT THE TWO EYES NEED TO INTERACT CHEMICALLY OR SYNAPTICALLY BUT WE'RE EXCITED IT CAN BE USED BECAUSE IT'S FAIRLY NONINVISITINGIVE AND IT'S AFTER THE LESION, SO YES WE DIDN'T OR NO, WE DIDN'T SEE AXONS GROW ALL THE WAY INTO THE BRAIN, WHEN WE JUST DID VISUAL STIMULATION AND WE THEED TO APPLY OTHER STRATEGIESAs WELL AND YOU CAN'T IMAGINE A NUMBER OF CONDITIONS INCLUDING GLAUCOMA AND CONDITIONS IN WHICH THE GANG LIAISON AN CELLS MAY NOT HAVE DEGENERATED BUT WERE MAYBE SICK OR MAYBE HEALTHY PEOPLE, TO INCREASE GANG LIAISON AN CELLS AND HEALTHY AND CONNECT SAID BECAUSE YOU SAW THE ROBUST EFFECT OF BLOCKING ACTIVITY. WE CAN'T FIND THEM IN THE EYE. SO SOME THING IS GOING ON THERE AND WE REALLY WOULD LIKE LITTLER GROUPS TO PICK UP ON THIS AND RUN WITH IT AS L. WE SAW TARGET SPECIFIC REENNERIVATION WHICH SUGGESTS THERE MAY BE UPREGULATION OF GUIDANCE CUES AFTER INJURY AND PARTIAL RESTORATION OF FUNCTION. ARE WHAT ARE MOLECAR MECHANISMS OF REGENERATION, WHAT ARE THE ROLE OF GUIDANCE MOLECULINGS. WE DIDN'T LOOK AT EVERY GANG LIAISON AN CELL TYPE. HOW ACCURATE IS THE WIRING, HOW DO WE THOSE TO GO BACK TO TARGETS. MAYBE SOME TRIED AND FAILED AND DIED WE DON'T KNOW. WHY DO THE REINNERIVATES AXONS RESTORE SOME BUT NOT OTHERS. THERE WILL BE AN IMPORTANT ROLE OF SYNAPSE FORMATION OR REFORMATION, I SHOULD SAY, VERY LITTLE IS KNOWN ABOUT HOW SYNAPSE FORM IN THE ADULT BRAIN AND SYNAPSE REFORMATION AND THAT'S ANOTHER ONE OF THE CALLED ARMS FROM THE AGI AND THEN WE HAVE TO GET FROM MOUSE TO HUMAN. WHETHER OR NOT THE PRIMATE SHOULD BE THE MARMA SET OR MACAQUE MONKEY, IT'S UP FOR DEBATE. WE WILL TALK ABOUT THAT, THERE'S JUST ONE SLIDE, ALEX WHO I MENTIONED EARLIER, AND I HAVE TEAMED UP AND LOOKING AT OR NOT THE SAME MOLECULES THAT ARE RESPONSIBLE FOR CONNECTING THE SPECIFIC GANG LIAISON ANS ARE ALSO TARGETS DURING DEVELOPMENT AND HE AND I HAD BACK-TO-BACK PAPERS AND THAT WAS IN 2015 SHOWING IN OUR CASE A ROLE FOR THE CONTACTINS, CELL ADHESION MOLECULE AND IN HIS CASE THE PLEXIN SEMIFORIN MOLECULES AND CONNECTING TARGETS IN THE BRAIN STEM. THIS IS VERY PRELIMINARY BUT IT'S REPEATED THIS A NUMBER OF TIMES SO HERE'S A SHAM CRUSH, CTB LABELING, ALL AXONS AND THIS IS A LEVEL, THE PROTEIN THAT'S VERY IMPORTANT FOR CONNECTING SPECIFIC GANG LIAISON AN DISELS TO THE TARGETS DURING DEVELOPMENT, IS KIND OF AT A LOW LEVEL, AND THIS IS STILL EXPRESSING IN AXONS BUT AFTER CRUSH, THEY GO UP LIKE GANG BUSTERS AT THE SITE, WE DON'T KNOW, BUT IT DOES SEEM THAT CRUSHING CELLS INJURYING THEM IS UPREGULATION AND THIS IS JUST HIGHER MAGNIFICATION, UPREGULATION OF THESE MOLECULES THAT WE KNOW DURING DEVELOPMENT WERE IMPORTANT FOR CONNECTING THOSE TO THE TARGET NEGLIGENCE THE FIRST PLACE, AND WE'RE VERY INCREEINGED BY THIS WHERE YOU WANT TO TEST THE FUNCTIONAL ROLES BY MISS EXPRESSION, TOPIC EXPRESSION, ONE CAN IMAGINE IN PATIENTS, POSSIBLY THROUGH A SERIES OF VERY SELECTIVE NEUROSURGERIES FAITHING BEADS OR APPLICATION OF GUIDANCE MOLECULES, PERHAPS COAXING BACK LARGE NUMBERS TO THEIR TARGETS IT WILL BE CHALLENGING BUT IF THAT'S THE BEST THING THAT IS AVAILABLE, AND YOU KNOW YOU CAN DO THAT PERHAPS MAYBE IT JUST TAKES ONE LONG DISTANCE ACCUSABLE CUE TO ATTRACT THESE AXONS WE NEED TO EXPLOAZ THESE IDEAS IN ANIMAL MODELS FIRST. AND NOW, THE QUESTION ABOUT HOW DO YOU FROM MOUSE TO HUMAN. SO THIS IS AN AREA THAT MY LAB STUMBLED INTO. I THINK YOU FIND YOURSELF BACK IN SILICONE VALLEY AND NEXT THING YOU KNOW YOU'RE WORKING ON VIRTUAL REALITY. AND THAT'S BECAUSE THESE TECHNOLOGIES ARE ADVANCING VERY FAST AND BECAUSE I PUT ON THE RESPONSE TO ONE OF MY COLLEAGUES SAYING, YOU HAVE TO TRY THIS AND DISCOVERED IT'S AN ALL-INCLUSIVE EVENTS AND KIDS WILL DIGS APPEAR INTO THESE GOGGLES AS TIME GOES ON BUT ADULTS - IT IS REMARKABLE, HOW IMMERSIVE IT IS. AND WE THINK ABOUT HOW WOULD WE DELIVER VISUAL STIMULATION IN A WAY THAT'S NOT MONOTONOUS AND MAYBE ROBUST AND MAYBE IN A MOVE OR EXPERIENCE THAT THEY WOULD WANT TO PARTICIPATE IN. SO WE THOUGHT ABOUT TESTING THE ROLE OF VISUAL STIMULATION USING VIRTUAL REALITY, COUPLE REASONS, ONE IT'S IMMERSIVE AND ENGAGING AND HARD TO DO IN AN AUDFORMAT LIKE THIS BUT PLANE OF YOU TRY IT AND I ENCOURAGE TO YOU TRY IT IF YOU GET THE CHANCE, THE OTHER THING THAT CAN YOU DO WITH VIRTUAL REALITY IS YOU CAN TRACK EYE MOVEMENTS AND MOVE THE VISUAL STIMULUS AROUND AND DOING THIS WITH THE VISUAL STIMULUS IN ORDER TO DIRECT THE STIMULUS TO THE RETINA IN WHICH YOU WANT TO CREATE ELEVATED LEVELS OF ACTIVITY AROUND LESIONS OR WITHIN LESIONS, SPECIFICALLY, AND THEN THE VISUAL RESTORATION FIELD HAS FOCUSED PREVIOUSLYOT QUESTION OF WHETHER OR NOT ENHANCING NEURAL ACTIVITY IN THE BRAIN OR EYE OR BOTH CAN IMPROVE VISION AND THERE ARE REPORTS HERE AND THERE ONE OF THE MAJOR CRITICISMS OF THAT EFFORT HAS BEEN THAT PATIENTS CHEAT WITHOUT KNOWING IT, THEY MOVE THEIR EYES AROUND A LOT. THEIR EYES ARE A CAPTIVE AUDIENCE AND THEY CAN CLOSE THEIR EYES BUT WE CAN TRACK IF THEY DO THAT. SO WE'RE NOW ENTERING, WE HAVE WRITTEN THE IRBEXPZ WE'RE HAVING THOSE IRBs REVIEWED OF DOING TESTING BASE LINE VISION IN A NUMBER OF DIFFERENT WAYS THIS IS A COLLABORATION WITH JEFF GOLDBERG, IN OPTICAL IMAGES THAT WILL MOLOGY. WE WILL TALK ABOUT IMMERSIVE. GENERALLY YOU ENTERTAIN PEOPLE AND KEEP THEM INCREEINGED NOT 10 HOURS A DAY BAH DRY IT IN A WAY REGENERATES THIS OR ENGAGES PLASTICITY AT THE LEVEL OF CORTEX WE DON'T CARE, WE JUST WANT TO TO SEE BETTER ANDEN EVALUATE IMPROVEMENT BECAUSE WE% DON'T HAVE RESULTS YET, IN A COUPLE OF YEARS I PROMISE WE WILL HAVE RESULTS ONE WAY OR THE OTHER. HOPEFULLY WITHIN 12 MONTHS, 24 MONTHS FOR SURE. AND I'M EXCITED THAT [INDISCERNIBLE] AGI MOVED TO STAND AFFORD AND--STANFORD AND USED DIFFERENT OPTICS RANGING FROM DYSTROPHYS AND LOOKING AT LETTER CODED VARIATIONS ON THE DIFFERENT STRUCTURAL FEATURES HE CAN IMAGE ALT HIGH RESOLUTION IN THE BACK OF THE HUMAN EYE AND SO GETTING WAY BEYOND SNELLING CHARTS AND THEN THE EYE EXAMINE AND THINGS LIKE HUMPHREYS EXAMS AND PUSHING WHETHER OR NOT CERTAIN FORMS OF VISUAL STIMULATION DIRECTED AT PARTICULAR LOCATIONS MAY BE IN A PATIENT THAT HAS, YOU KNOW SPECIFIC TYPES OF LESIONS, OR FEATURES WITHIN THE EYE, DIRECTING VISUAL SIMULATION RIGHT THERE, AND VR IS A TERRIFIC WAI TO DO THAT, VERY HARD TO DO IN OTHER FORMATS. OKAY, SO I THINK IN THE LAST COUPLE OF MINUTES, I WANT TIME FOR QUESTIONS, I WILL GIVE A TEASER OF WHERE VR CAN ALSO BE USED SO RATHER THAN GO INTO THE ANIMAL DATA I WILL QUICKLY SHOW YOU TWO THINGS, AND I'M MINDFUL OF THE TIME. OKAY? SO, I WANT TO KNOW WHY THIS IS CARRY AND IF I HAD SMOAD YOU SOMETHING ELSE, IT MIGHT NOT BE AND IT HAPPENS EXTREMELY FAST AND I WANT TO KNOW WHERE IN THE BRAIN AND HOW IN THE BRAIN IT HAPPENS AND AT RISK OF GETTING INTO A WHOLE OTHER CONCEPT TRAIN, I THINK IT'S INCREDIBLY INTERESTING QUESTION AND VR HAS A ROLE TO PLAY AND VISION HAS A ROLE TO PLAY. A LOT OF PEOPLE HAVE STUDIED FEAR BUT NOT MANY PEOPLE HAVE ADDRESSED WHAT ABOUT VISUAL STIMULI SPECIFICALLY IS SCARY. AND SO, THIS IS A SIMPLE EXAMPLE OF HOW BRIGHTNESS, YOU WOULD WALK DOWN THIS I DON'T THINK YOU WANT TO WALK DOWN THIS, YOU WOULD BE LESS LIKELY, I WOULD PICK THIS FRANKLY. THIS USUALLY GENERATES THE AWWWW-REFLEX, I DON'T WANT TO TOUCH THIS DOG, SO I WILL ILLUSTRATE THE ESSENCE OF THE PROBLEM HERE, AND ALSO, THINGS LIKE UNJUSTICE ACCESSALATION, THIS IS ENOUGH TO GET PEOPLE'S HEART RATE GOING, RIGHT? WHAT IT IS IS INTERESTING, IT'S--THIS IS A BEANIE BABY THAT'S COMING, I DIDN'T WANT TO SHOW YOU THAT, I THOUGHT WE WOULD END ON A LIGHTER NOTE, BUT THE UNDULATION AND MOVEMENT ARE ALSO VERY INTERESTING FEATURES, RIGHT? AND SPEED AND VECTOR ARE OFTEN WHAT'S SCARY. UNDULATION LEADS TO UNPREDICT DIBL ABOUT WHAT DIRECTION SOMETHING IS GOING TO GO AND IT CAN MOVE FAST AND QUICKLY IN ONE DIRECTION, THAT'S SCARE EXPE THERE IS GOES UNFORTUNATELY THE BEANIE BABY DOESN'T MAKE IT, THE OKAY, YEAH, SO, JUST VERY BRIEFLY AND I WILL TRULY SKIP OVER THE SLIDES BECAUSE I'M HAPPY TO TALK ABOUT THIS OFFLINE, BUT THIS IS--LET ME JUST SHOW THE BEHAVIOR THAT WAS REALLY BEAUTIFULLY SET UP FOR--THERE'S BEEN A LOT OF DISCUSSION, THIS DOVETAILS ON REGENERATION IN A WAY, VISUALIZATION IS HARD TO PROBE BUT SOME ARE ROBUST, IT WAS DISCOVERED BY A GRADUATE OF MARCUS M, ISTER 'S LAB, AND NOW SHE'S IN MY LAB, SHE'S STUDYING VISUAL FEAR IN HUMANS THIS, IS THE FIRST TIME AND EVERY TIME VISUAL LOOM IS PRESENTED. IT GETS SCARED AND LOOK AT, THIS IS AN ACTIVE PROCESS, IT ALMOST LIKES LIKE IT FROZE IN THE MOVIE OR THE MOUSE MIGHT DO THIS. RUN, HIDE. NOT SO HIDDEN I'M GOING TO GET% OUT OF HERE, OKAY? ONE OF THE TWO EVERY SINGLE TIME SO I DON'T WANT TO GET INTO THE DETAILS OF ALL THIS, WHAT I WILL TELL YOU IS TWO TEASERS AND THEN WE WILL MOVE TO QUESTIONS WE HAVE A LOT OF MECHANISM MECHANISM--MECHANISTIC DATA BUT FORGIVE ME FOR SKIPPING A LOT OF THIS BUT LINDSAY HAS IDENTIFIEDA I NUCLEUS CONNECTED TO THE PATHWAY THAT WHEN STIMULATED CONVERTS FEAR INTO WHAT IS ARGUABLY AGGRESSION, TAIL RATTLING IN MICE IS WHEN YOU PUT THEM TOGETHER AND FIGHT, IT'S LIKE CHEST THUMB NOTHING PRIMATES OR GUYS IN BARS THAT HAVE A FEW TOO MANY AND GO AT AND NOT A GOOD DECISION, RIGHT? SO HERE SHE'S STIMULATING THE THAM ALIC AREA, WITH CHEMICAL OR CAN DO THIS WITH OPTICAL IMAGES O GENETICS IT FREEZES FOR A MOMENT AND THEN IT WALKS AROUND AND TAIL RATTLING IF IT'S NOT AGGRESSION, MAKE THIS IS MOUSE MORE SALIENT TO THE STIMUE LUGZULOUS, THIS MOUSE IS COMFORTABLE STROLLING AROUND AND RATTLING ITS TAIL AGAINST THE WALL AND IT WILL CONTINUE TO DO THIS OVER AND OVER AGAIN. AS LONG AS SHE'S STIMULATING THIS NUCLEUS, I'M INCREEINGED BY THIS, HOW AND WHY IS IT THAT THE CERTAIN SENSATIONS WITH A SINGLE VISUAL PATHWAY CAN CONVERT ONE SENSATION INTO ANOTHER KIND OF OF EMOTIONAL RESPONSE, BEHAVIORIAL RESPONSE. SO I DON'T WANT TO SPEND TOO MUCH TIME ON THIS BUT WHAT I WILL TELL SURVEYS THAT WE'RE BACK NOW V. R. BECAUSE WE'RE USING THIS ANN EMERCKIVE WAY TO PROBE VISUAL FEAR IN HUMANS AND ACTUALLY THIS IS STEVE BECKER, HE'S OWLLY ON--UP ON A PLATFORM, MANY METERS ABOVE THE GROUND, HE'S ON THE FLAT OF MY LAB. THIS IS MICHAEL MULLER, IF YOU EVER SEEN THIS CELEBRITY PHOTOGRAPH OR MBBA OR OLYMPIC PHOTOGRAPH, HE TOOK IT AND MULLER HAS A THING FOR COLLECTING MOVIES AND HE LIKES SHARKS SO HE AND HIS BUDDIES LIKE TO SWIM OUTSIDE THE CAGE IN GUADALUPE, HE LIKES TO DRESS LIKE A TOWN - SO THEY COME CLOSE TO HIM AND THEY WENT AND GOT US IMMERSIVE V. R. FOOTAGE AND HERE STEVEN'S NOT LOOKING AT THE SHARK THING I'LL USE HIS BODY LANGUAGE, I'LL TURN OFF THE SOUND AND IS--YOU KNOW THIS IS HIS RESPONSE, HE THINKS HE'S ABOUT TO FALL OFF A BRIDGE. LET ME SEE IF I CAN TURN UP THE SOUND, MAY I DO THIS? I'LL ASK PAUL? CAN I--HE AGREED TO THIS. SO I DON'T WANT THIS TO GO TOO LEWD. >> LET ME IF I CAN. >> OH MY GOD--SO THAT'S ESSENTIALLY, THE OH MY GOD RESPONSE. NOT AN UNUSUAL ONE TO FEAR. SO, WHAT'S INTERESTING ABOUT THAT TO ME IS THAT IT'S SO IMMERSIVE, HE KNOWS HES IN THE LAB AT SOME LEVEL BUT THE BRAIN THINKS HE'S UP ON THE PLATFORM AND THERE'S NOTHING UNUSUAL ABOUT THAT RESPONSE. EVERYONE THINKS THEY'RE WITH THE SHARKS WHEN YOU PUT THIS ON AND JUST TO ILLUSTRATE, YOU CAN IMAGINE DOING A VISUAL PARADIGM, MAYBE NOT SCARING PEOPLE BUT WALKING THROUGH THE SYDNEY OPERA HOUSE IN VIRTUAL REALITY, I TOOK OFF THE GOGGLES ON THAT ONE AND I ACTUALLY WANT TO GO TO THE REAL OPERAEROUS AND I DON'T WANT TO SPOIL THE EXPERIENCE, SO IT'S SO IMMERSIVE AND A PORTABLE CLINIC AND WE'RE THINKING PROBE AND STIMULATE REGENERATION IN THE VISUAL SYSTEM AND LAST BUT NOT LEAST, THIS IS A NEW COLLABORATION WHERE IT'S GREAT TO RECORD BEHAVIORIAL RESPONSES AND SOPHISTICATEDMATIC RESPONSES AND GALVANIC SKIN RESPONSE AND ECG AND THINGS LIKE THAT FROM SO CALLED NORMALS OR HEALTHY INDIVIDUALS LIKE STEPHEN OR BUT IT'S ALSO GREAT OPPORTUNITY NOW THAT EDDIE CHANG, COLLEAGUE UP UP AT UCSF, THEY HAVE PATIENTS HAVE ELECTRODES EMBED INDEED THIS SIGNALS AND THESE ARE SIGNALS WHILE A PATIENT VIEWS FROM THE AMYGDALA FROM A GREAT WHITE SHARK AS IT'S PASSING THROUGH HIS VISUAL FIELD SO WE CAN GET NEURAL EVENTS TO SOPHISTICATEDMATIC RESPONSES TO BEHAVIORIAL RESPONSES AND THEN SUBJECTIVE REPORTS AND THAT'S WHERE WE'RE HEAD INDEED A MAJOR WAY IN THE NEXT COUPLE OF YEARS YEARS AND THESE EFFORTS REGENERATION AND VISUAL FEAR ULTIMATE GOAL MAYLY AS A WAY TO PROBE THINGS LIKE ANXIETY AND PTSD ARE WHERE I WILL PUT MOST OF MY LAB'S EFFORTS, WE WILL CONTINUE WITH THE DEVELOPMENT WORK OF COURSE. SO I WILL FINISH THERE. I THINK WE'RE ON THE HOUR. SO I WANT TO THANK THE PEOPLE WHO KRIEWBILITY TO THIS WORK, LINDSAY SALAY, FABULOUS GRADUATE STUDENT. STAY TUNED TO THE MECHANISTIC FEATURES OF CONVERTING SCARED MICE INTO BRAVE MICE. ALBERT LIM, AND CURRENTLY AT GENENTECH, AND THEN REGENERATION IF THE BRAIN, CORTICO PLASTICITY, MELIS YILMAZ, IN THE LAB, WORKING ON THE THESE AND I'M VERY GRATEFUL TO THESE FOLKS AND THE FOUNDATIONS FOR FUNDING AND NONE OF THE WORK WE COULD HAVE DONE WOULD HAVE HAPPENED WITHOUT THE SUPPORT OF THE NEI AND NIH. THANK I SO MUCH AND I APPRECIATE YOU--THANK YOU SO MUCH AND I APPRECIATE YOUR ATTENTION. [ APPLAUSE ] I WILL BE HAPPY TO ANSWER QUESTIONS BUT I REALIZE SOME MAY HAVE TO LEAVE GIVEN THAT WE'RE ON THE HOUR. YES? >> SO PATIENTS WHO ARE [INDISCERNIBLE] THE MTOR MIGHT BE [INDISCERNIBLE] [INDISCERNIBLE] SO THE QUESTION IS WHETHER OR NOT IN GLAUCOMA PATIENTS WHO ARE LOSING GANG LIAISON AN CELLS WHETHER THE MTOR THERAPY WOULD HELP SLOW THAT LOSS OR REGENERATE GANG LIAISON AN CELLS. I'M NOT A CLINICIAN BUT I WILL SAY THIS, THE ANIMAL DATA SAY YES, I THINK IT'S IMPORTANT TO REPLICATE THESE RESULTS FIRST DISCOVERED BY KEVIN PARK AND OTHER SAYS HAVE REPLICATED, OF COURSE, IN A PRIMATE MODEL. I THINK IT WILL BE VERY IMPORTANT. OF COURSE THERE'S EFFICACY AND SAFETY ISSUES RELATED TO INCREASING MTOR BECAUSE THE CANCER BIOLOGIST MIGHT LEAP OUT OF THEIR SEATS AND COMUP TO THE PODIUM IF I SAY LIKE LET'S PUT IN RHEB VIRUS BECAUSE THE CELLS GROW VERY LARGE, WE DO KNOW THEY MAINTAIN THEIR RECEPTIVE FIELDS SO PROVIDED YOU CAN BYPASS OTHER DELETERIOUS POTENTIAL EFFECTS OF INCREASING MTOR, YES. I BELIEVE THAT THAT'S A PATHWAY THAT'S WORTH LOOKING INTO AND SERIOUSLY IN THE CLINICAL REALM. WHETHER OR NOT IT'S THROUGH PEP TIDES OR WHETHER OR NOT IT'S GOING TO BE THROUGH A GENETIC STIMULUS, MAYBE YOU WANT MTORON FOR A FEW HOURS EACH DAY AND THE GENETIC TEALS THAT ARE AVAILABLE NOW ALLOW YOU TO DO THESE KIND OF SHORT DURATION PULSES OF GENE ACTIVATION, BUT WE--I THINK THE PRIMATE MODELS WILL GIVEN THE SIMILARITIES OF THE EYE AND ARCHITECTURE AND LONG DISTANCES OF COURSE THEY HAVE TO GO THROUGH THE STANDARD TRIALS BUT I WOULD PUT MY MONEY ON IT IN TERMS OF HAVING A STRONG EFFECT AND WHETHER OR NOT CAN YOU AVOID SIDE EFFECTS IS THE KEY ISSUE. IN THE CRASH MODEL YOU LOSE THE AXONS, SO IN MOST HUMAN DISEASES AND MOST HUMAN DISEASES THAT EFFECT THE OPTIC NERVE, THAT CLASS IS MORE GRADUAL. SO IS THERE A POSSIBILITY THAT THE REMAINING AXON SCAN HELP IN THIS REGENERATION PROCESS? >> GREAT QUESTION. SO, YOU KNOW THERE WAS A LOT OF DEBATE ABOUT WHAT MODELS TO USE IN GLAUCOMA, DBA-TWO-J, BEAD MODEL, PRESSURE. IT'S INTERESTING. THE FIELD FROM THE PAPERS I REVIEW AND SEE AND SUBMIT IS KIND OF SAID, YOU KNOW THEY ALL HAVE THEIR VALUE. SO THE I THINK THAT'S A GOOD PROGRESSION, WE NEED MORE PEOPLE IN THE FIELDS AND NEED TO USE MORE MODELS, PEOPLE ARE SHARING REAGENTS MORE AND PEOPLE ARE SENDING PEOPLE BETWEEN LABS, WE CERTAINLY TAUGHT PEOPLE THE BEAD MODEL AND WE LEARNED IT FROM OTHERS AND THERE'S MORE SHARING WHAT HASN'T HAPPENED YET, CULTURALLY IF YOU WILL. AND I WOULD LIKE TO SEE MORE OF IS INDIVIDUAL LABS USING MULTIPLE MODELS, SO WE USE THE BEAD MUDDLE AND CRUSH MODEL BUT WE HAVEN'T USEDDED BEAD MODEL IN REGENERATION BUT WE'RE DOING THAT NOW. THE CRUSH MODEL HAS THE DRAWBACKS BUT TO ANSWER YOUR QUESTION MORE DIRECTLY. THE PHYSICAL PRESENCE OF OTHER AXONS WE KNOW DURING DEVELOPMENT ACTS AS A GUIDE FOR SUBSEQUENT AXONS TO USE THIS, THE PIONEERING EFFECT OR AXONS AND EVERYTHING WE SEE FROM THE ROLL OF ACTIVITY TO THE ROLL OF MTORTO--I'M DRAWING A BLANK CSPG REGULATION THAT WE SEE DURING DEVELOPMENT AS A POSITIVE IMPACT OF GROWTH OF GANG LIAISON AN CELLS HAs A POSITIVE IMPACT OF REGENERATION OF GANG LIAISON AN CELLS SO PHYSICAL SUBSTRATES FOR GROWTH MATTER DURING DEVELOPMENT BUT I WILL BE WILLING TO BET THAT THE PHYSICAL SUBSTRATE FOR GROWTH WILL HELP IN THE ADULT AND I JUST HOPE SOMEONE DOES THE EXPERIMENT. YOU KNOW WE WILL DO, HOW DO I SAY THIS. I WILL SAY THE WAY IT IS WE WILL DO AS MANY EXPERIMENTS AS I CAN MAY MY PEOPLE TO DO WELL BUT THERE'S A THRESHOLD TO THAT. >> THANK YOU. >> THE FUNDING AND THE WELL PART SO WE NEED MORE GROUPS WORKING ON THESE THINGS. THANKS FOR THE QUESTION. >> HI, YOU MENTIONED GLIAL SCAR REAL QUICKLY, HOW IMPORTANT DO YOU THINK THE GLIAL SCAR IS IN THE OPTIC NERVE CRUSH MODEL IN STOPPING AXONAL REGENERATION. >> THE WORD WORD ANSWER IS VERY. IT'S INTERESTING THAT INTRINSIC STIMULATION ALONE, YOU CAN GET AXONS REGENERATING THROUGH LESION SITES BUT NOT INTO THE BRAIN. BUT THEY DO REGENERATE BUT NO ONE HAS DONE THE COMBINED NEUTRALIZATION OF A GLIAL SECRETED FACTOR AND MTOR, TO MY KNOWLEDGE, FORGIVE ME IF I'M MISSING A PUBLISHED REPORT. PLEASE SEND IT TO ME IF I MISS TODAY, IN CONJUNCTION WITH THE ANGRY E-MAIL I USUALLY GET BUT I WANT TO KNOW THE LITERATURE. IF IT'S OUT THERE, I APOLOGIZE, SO I DOABILITY THINK IT'S BEING DONE EMPLOY BUT THAT'S AN IMPORTANT PONENT, BUT THEY REALLY HAVE--THEY GOT PEOPLE TOGETHER AND SAID WHAT ARE THE CRITICAL ISSUES AND WE DON'T KNOW THE FUNCTIONAL ROLE OF GLIAL SCAR WHEN COMBINED WITH INTRINSIC GROWTH PROMOTERS BUT WE HAVE TO FIGURE THAT OUT. >> DO PEOPLE KNOW WITH WEATHER THERE'S GLIAL SCAR FORM NOTHING THE CRUSH MOUSE MODEL. >> YTS IT'S, YES IT'S ALSO SETTING UP A PHYSICAL BARRIER. NOT WORK WE'VE DONE BUT OTHER GROUPS INCLUDING GROUPS HERE. >> HI,. >> HI,. >> IM CURIOUS IN YOU LOOK AT THE HEALTH OF THE TARGETING SYSTEMYS IN SO IF YOU LESION AND GET RID OF THERE--RETINAL GANG LIAISON AN CELLS AND THEN THE DOWN STREAM EFFECT OF THE HEALTH OF THE TARGET FOR AS A POTENTIAL REASON FOR WHY IT'S NOT REFORMING. >> GREAT QUESTION. AND THE SHORT ANSWER IS WE HAVEN'T, VERY FEW PEOPLE HAVE. THERE'S A REAL DIRTH ROLE OF WORK LOOKING AT THESE. WHEN YOU DENERVATE A TARGET DEVELOPMENTALLY. GOING BACK TO DEVELOPMENT. WE GET ANY CLUES IT'S THE FOLLOWING, IF YOU DENERVE A TARGET DEVELOPMENTALY, THE SYNAPTIC TRANSMISSION IS ONE OF THEM AND THE NEURON THERE IS SHRINK THEIR OUTPUTS DEGENERATE TO SOME DEGREE. THIS IS THE LATE LARRY CATS DID BEAUTIFUL WO, ON THIS. HE SHOWED IF YOU DEPRIVE THE NUCLEUS OF INPUT BUT YOU SUPPLY IT WITH GROWTH FACTOR WITH THE PROJECTION THROUGH THE CORTEX, PUTTING BEADS IN AND THAT THINGS OF THAT SORT, YOU CAN PRESERVE THE GENERICULATEAND THERE IT WAS ALSO SHOWN THAT IF YOU CLOSE ONE EYE, THERE'S A SHRINKING OF THE DEPRIVED EYE LIERS, BUT IN THE NONDEPRIVED EYE LAYERS SO WE NEED MORE WORK BUT ALMOST CERTAINLY YES. >> HI, SO, I'D BE CURIOUS TO HEAR YOU SPECULATE A BIT MORE ABOUT THE MECHANISM BY WHICH CONTRA LATERAL DEPRIVATION IS SIGNALING SO FOR EXAMPLE, IF YOU WERE TO TAKE SAY, A CHUNK OF RETINA AND OPTIC NERVE THAT ARE STILL ATTACHED, AND PUT THEM INTO A HOST, DO YOU THINK THAT THERE WOULD BE--YOU KNOW, IN THE PERO NEUROECTODERMAL UMKC OR VENTRICLE OR CIRCULATING CUES ESSENTIALLY. ARE YOU SEEING SOMETHING LIKE THAT? OR'RE YOU THINKING IT'S INFORMATION COMING ALONG THE PATHWAY? ET CETERA? >> I HAVEN'T THOUGHT ABOUT WHAT YOU'RE DESCRIBING BUT TRANSPLANTATION CAN REVEAL A LOT. JEFF GOLDBERG DID A EXPERIMENT THAT GETS AT THIS A BIT. HE TOOK REALLY YOUNG CELLS THAT ORD NARRLY WOULD GROW IN A DISH OR INVIVO. AND IN HIS SCIENCE PAPER BUT IT WAS SUPPLEMENTAL DATA EAR HIDDEN IN THE KIND OF END OF PAPER, IT WAS A CRITICAL EXPERIMENT AND HE PUT IT INTO THE ADULT CORPUS CO LOSS UMKC. THE MILLUE IS TERRIBLE FOR REGENERATION AS FAR AS WE KNOW. AND THEY EXTENDED AT A FAST GROWTHERATE. SO IT'S CREATING A PERIPHAL LIKE THING EXCEPT IN REVERSE, YOU TAIB A BABY NEURON AND PUT IT IN A BAD ENVIRONMENT AND IT GROWS. SO THAT SUGGESTS THAT IT'S SOMETHING REALLY INTRINSIC TO THE NEURONS THAT'S ACTIVATED BUT IN THIS ARRANGEMENT OF BIASED VISUAL STIMULATION, MTORAND SO FORTH, IT'S HARD TO IMAGINE MANAGE THAT'S BASED ON LOCAL ENVIRONMENT, THE DATA KIND OF POINT TO SOMETHING DIFFUSIBLE BUT NOT PURELY DEFUSE ABLE BECAUSE WE DID EXPERIMENTS IN WHICH ALBERT REVERSED THE DIRECTION OF TREATMENT SO HE'S CLOSING NOW THE TREATED EYE AND HE'S INJECTING THE OTHER EYE AND HE DIDN'T SEE THE SAME REESKT EFFECTS SO WHAT I THINK WE NEED TO DO IS SOME NICE RNA SEEK, GROWTH STIMULATE GANG LIAISON AN CELLS AND FORTUNATELY, WE HAVE SUPPORT TO DO THAT. THERE'S A GRANT THAT INVOLVES HOLLY KLEIN, JEFF GOLDBERG AND LARRY BENOWITS AND LOOKING AT THESE TRANSSCRIMENT OHMS OHMS AND PROTEIN COMPLEX I DON'T MEANS AND WE WON'T NECESSARILY GET THE CUE THAT WAAND MIGHT GET THE RECEPTOR OR SOME HINTS INTO WHAT THE QR. IF ANY OF HAVE YOU IDEAS ABOUT THIS OR WANT TO PURUE THE LINE OF EXPERIMENTS THIS WILL NOT BE A SINGLE LAB OR QUADLAB EFFORT. ONE BRIEF POINT IN THE SPINAL CORD, THERE'S INCREEINGING LITERATURE THAT SUPPORT THIS IS IDEA OF BIASED USE. ONE OF THE BEST TREATMENTS FOR GETTING PLASTICITY IS TO TIE UP THE GOOD LIMB AND FORCE USE OF THE IMPACTED LIMB TO DRIVE--THE OLD WHERE SOME OF YOU MAY RECALL, I REALLY LOVE THIS STUFF, TIM SHALLERT, WHO WAS THE UT AUSTIN AND POPULARIZED THE EFFECT OF TYING UP THE GOOD LIMB AND THEN THAT WAS PICKED UP ON BY A NUMBER OF DIFFERENT LABORATORIES INCLUDING MARTIN SCHWAB'S LAB. I WILL SKIP BACK AND NORTH FOR THE LAST TWO QUESTIONS. >> HAS HAS [INDISCERNIBLE] BEEN TRIED IN FISH NEURONS WHERE THEY CAN REGENERATE THE EFFECT OF MTOR, P10, ET CETERA? >> NOT TO MY KNOWLEDGE AND YOU KNOW OF COURSE IN FISH THEARKS ADDING NEW NEURONS ALL THE TIME. >> LIKE A KNOCK OUT OF MTOR. I DON'T WANT TO PUT HER UP TO AN EXPERIMENT THAT SHE'S NOT DOING FOR WHATEVER REASON YOU BUT I THINK HOLLY IS DOING THESE EXPERIMENTS BECAUSE OF COURSE SHE HAS SUCH BEAUTIFUL DECADES OF BEAUTIFUL WORK IN THE FISH AND XENAFIS, AND THERE YOU WOULD EXPECT THAT MTORWOULD BE HIGH IN THESE BABY NEURONS AND THAT MIGHT BE THE REASON WHY THEY CONTINUALLY REGENERATE. THAT QUESTION IS STILL SO FUNDAMENTAL WE DON'T KNOW WHY THESE COLD BLOODED VERTEBRATES REGENERATE BUT IT'S SCREAMING MTOR. SO THAT'S EXACTLY THE RIGHT QUESTION. YEAH. >> THANK YOU. >> WE HAVEN'T DONE IT. I'M TRYING TO KEEP THE NUMBER OF SPECIES TO--ALTHOUGH IT WOULD BE EASIER TO WORK ON FIB THAN HUMANS BECAUSE IT TURP OTHER THAN OUT THAT'S COMPLICATED [LAUGHTER] >> I WAS CURIOUS IN YOU HAVE ANY SENSE OF IF YOU CATERED OR IF YOU TAYLOR THE STIMULUS TO WHAT MAYBE SOME TYPE OF GANG LIAISON AN CELL FIRST, WOULD YOU RECRUIT MORE OF THOSE TYPES OF GANG LIAISON AN CELLS, LET'S SAY IF YOU HAVE MORE LOOMING, WOULD YOU GET MORE LUMENS, OR CELLS TO GO BACK TO THE BRAIN OR IN A-- >> GREAT QUESTION. THE EXPECTATION IS YES, BECAUSE THOSE Ls ARE GOING TO SPIKE MORE IN RESPONSE TO A PARTICULAR STIMULUS SO WE'RE DOING THOSE. YOU KNOW IT'S VERY PRELIMINARY BUT WE THINK THAT THIS IDEA THAT ONLY SOME GANG LIAISON AN CELLS CAN REGENERATE BECAUSE OF SOMETHING THAT'S SPECIAL ABOUT THEM MOLECULARLY, IT'S TRUE, BEAUTIFUL WORK FROM JOSH AND OTHERS TO SHOW THAT THE ALPHA CELLS REGENERATE PARTICULARLY WELL AND THAT HAS TO DO WITH THE EXPRESSION OF IGF-ONE. BUT IN OUR TREATMENT, WE SAW RECOLLECT CELL TYPES THAT REGENERATE AS WELL. AND WHEN WE START RECRUITING THEM THROUGH VISUAL ACTIVATION, THIS IS ANOTHER REASON WHY WE REALLY WANT TO TRY THIS IN HUMANS AND TAYLORS THE TYPES OF VISUAL DEFICITS DEFICITS AND THOSE DEFICITS THAT THE PEOPLE HAVE, WE THINK THAT'S GOING TO BE THE CASE, AND AGAIN, VR IS A PERFECTLY GOOD WAY. WE KNOW THESE PEOPLE ARE DEFICIENT IN MOTION PERCEPTION AND WE THINK THAT IT'S MOTION PERCEPTION DUE TO LOSS OR INJURY OF GANG LIAISON AN CELLS IN THE VISUAL FIELD. VR IS THE PERFECT PLATFORM TO DELIVER TO THOSE LOCATIONS OR AROUND THE LESION, PERHAPS YOU WANT THOSE OR BI POLEAR CELLS CAN RECRUIT THE MOTION SENSITIVE CELLS. OR MOTION SENSITIVE CELLS AND ALL THAT CAN BE DONE IN VR IN A STRAIGHT FORWARD WAY, DIDN'T REQUIRE DEVELOPMENT OF A DRUG TARGET OR TOOL, SO WE'RE EXCITED TO PURSUE THAT. >> OKAY, THANK YOU ANDY YOU STIMULATED A LOT OF CONVERSATION HERE, THANK YOU FOR COMING. GOOD LUCK WITH MTOR, AND ALL OF THE MOLECULES THAT WE WILL BE USING IN PEOPLE SOMEDAY, THANK YOU.