WELCOME TO SECOND DAY OF THE BRAIN MEETING IT'S MY PLEASURE THIS MORNING TO INTRODUCE DR. FRANCIS COLLINS TO GIVE US WELCOMING REMARKS AND TO INTRODUCE OUR KEYNOTE SPEAKER. DR. COLLINS WAS APPOINTED IN 2009 TO BE THE 16th DIRECTOR AT THE NATIONAL INSTITUTES OF HEALTH AND RESELECTED BY PRESIDENT TRUMP TO CONTINUE IN THAT REGARD. BUT DR. COLLINS IS NOT JUST THE NIH DIRECTOR. HE IS FIRST AND FOREMOST A SCIENTIST AND BEST KNOWN FOR HIS WORK IN IDENTIFYING AND CLONING THE CYSTIC FIBROSIS GENE. AND THE FACT THAT HE IS A SCIENTIST THAT LED HIM TO TAKE ON THE JOB OF SHEPHERDING THE BRAIN INITIATIVE THROUGH THE INITIATION AND ESTABLISHMENT AND IT'S HIS EXPERIENCE WITH THE HUMAN GENOME PROJECT THAT LED HIM TO RECOGNIZE THAT IT WILL BE THE TOOLS THAT WILL HELP US UNDERSTAND THE BRAIN. SO DR. COLLINS LIKE MANY OF US WHO HAVE BEEN ATTENDING THE MEETING HAS BEEN AWED WITH THE PROGRESS WE'VE MADE BUT ALSO IMPATIENT FOR THE NEXT GENERATION OF DISCOVERIES. WITH NO FURTHER ADO I'LL INVITE DR. COLLINS TO TAKE THE PODIUM. [ APPLAUSE ] >> WELL GOOD MORNING TO ALL OF YOU. THANKS, JOSH. IT'S WONDERFUL TO HAVE A CHANCE TO BE HERE WITH YOU THIS MORNING TO SAY A FEW THINGS ABOUT BRAIN INITIATIVE. ONE OF THE MOST EXCITING THINGS HAPPENING AT NIH RIGHT NOW. SORRY THAT I WAS NOT ABLE TO BE HERE YESTERDAY. WE HAD AN ALL DAY REFRUIT AT THE HUMPHREY BUILDING WHERE THE MAIN HEADQUARTERS IS LOCATED. THE TOPIC WAS THE OPIOID CRISIS SO ALL OF THE THINGS THAT YOU'RE DOING TO FIGURE OUT CIRCUITS IN THE BRAIN THAT MIGHT ENABLE US TO COME UP WITH DIFFERENT STRATEGIES OR BETTER YET TO PREVENT IT TO TREAT CHRONIC PAIN -- DON'T HOLD BACK. IT'S CONSUMING THE ATTENTION OF A GREAT MANY PEOPLE ACROSS MANY DIFFERENT SECTORS RIGHT NOW AND WELL IT SHOULD. AND I REGRET THAT I WON'T BE ABLE TO SPEND MUCH TIME WITH YOU TODAY OR TOMORROW EITHER BECAUSE WE HAVE TOMORROW THE CONGRESSIONAL HOUSE APPROPRIATION SUB COMMITTEE HEARING FOR NIH BUDGET FOR FY19. BUT I THINK YOU WANT ME TO BE THERE AND ARTICULATE AND WELL PREPARED BECAUSE THIS IS THE MOMENT -- [ APPLAUSE ] >> WE ARE ON A ROLL ACTUALLY. IF YOU'VE BEEN PAYING ATTENTION THE CONGRESS OF THE UNITED STATES HAS SEEN IT AS A HIGH PRIORITY TO BRING NIH BACK INTO A STABLE UPWARD TRAJECTORY WITH A $3 BILLION INCREASE IN '18 AND WE HOPE FOR GOOD THINGS IN '19 WITH SOME OF OUR HEROES TALKING ABOUT HOW IMPORTANT IT IS TO MAIN THIS EVEN IN A DIFFICULT BUDGET ARENA. I'M THRILLED TO STILL BE THE NIH DIRECTOR. YOU HEARD THE INTRODUCTION. THAT MAY NOT HAD BEEN EXPECTED. ESPECIALLY WHEN THERE WAS A CHANGE IN PARTIES SO I THOUGHT WHEN WE CAME TO THIS POINT I MIGHT BE HERE BROWSING AND ENJOYING THE WHOLE THING IN A DIFFERENT WAY BUT I'M PRIVILEGED AND DELIGHTED TO CONTINUE TO TRY TO STEER THIS AMAZING SHIP AND PARTICULARLY TO TRY TO NURTURE SOME OF THE GROUNDBREAKING PROJECTS THAT INCLUDE THE BRAIN IN A VERY BIG WAY TO SHED ENTIRELY NEW LIGHT ON FUNDAMENTALS ABOUT HOW LIFEWORKS AND DISEASE HAPPENS. I WANT TO SAY A LITTLE BIT ABOUT WHERE WE'VE COME FROM AND WHERE WE'RE GOING. I'VE ENJOYED LEARNING ABOUT THIS WHOLE ENTERPRISE. EVERY TIME I THINK I KNOW WHAT IS GOING ON YOU COME UP WITH SOMETHING NEW BUT THAT'S FINE. KEEP STRESSING ME. I LIKE THAT. SO IT WAS FIVE YEARS AGO IN THE WHITE HOUSE WE HAD THIS ANNOUNCEMENT OF THE NEXT GREAT AMERICAN PROJECT. THE BRAIN INITIATIVE AND LEARNING THE MESSAGE OF THE BRAIN AND WITH A GREAT DEAL OF EFFORT ON A PART OF A WORKING GROUP PUT FORWARD THIS PLAN FOR BRAIN 2025 FOCUSED ON CIRCUITS AND NETWORKS AND A GREAT DEAL OF FOCUS IN THE EARLY FIVE YEARS ON DEVELOPING TECHNOLOGIES. THAT ALLOW US TO SEE WAS HAPPENING IN THAT CRITICAL SPACE IN BETWEEN WHOLE BRAIN IMAGES AND SINGLE NEURON IMAGES. WE NEED TO UNDERSTAND IN GREAT DETAIL INCLUDING WHAT IS HAPPENING IN REALTIME SO WHERE HAVE WE COME FROM WHEN YOU LOOK TO SEE THE NUMBER OF THINGS THAT HAVE HAPPENED, IT'S PRETTY BREATHTAKING. I'M NOT GOING TO GO THROUGH ALL OF THE THINGS ON THIS PARTICULAR IMAGE BUT I THINK YOU CAN SEE STARTING THERE IN APRIL 2013 WITH THE REPORT THE FIRST AWARDS IN SEPTEMBER 2014 AND THEN GOING FORWARD LET'S COME UP HERE TO DECEMBER 2016 THE THIRD BRAIN BI MEETING AND ALSO AN ANNOUNCEMENT THAT THERE WOULD BE STABLE FUNDING FOR THE PASSAGE OF THE CURE ACT. SOMETHING THAT DOESN'T USUALLY HAPPEN. USUALLY WE HAVE TO DO EVERYTHING YEAR BY YEAR SO TO SEE THAT PUT FORWARD AND I'M SHOW YOU THE DOLLAR FIGURES GIVES YOU THE CONFIDENCE THAT THIS IS A PROJECT THAT WE CAN INVEST IN AND THAT REFLECTS THE WAY INCH THE CONGRESS SEES THIS AS A FUNDAMENTALLY TRANSFORMING EXCITING PROJECT. SO HERE WE ARE NOW APRIL 2018 WITH THE 4th BRAIN BI MEETING IT TERMS OF THE A WOODS -- WORDS OVER THE YEARS THE COLORED PENS ARE INDICATED BY WHICH YEAR THE AWARD WAS MADE AND YOU CAN SEE THERE ARE A LOT OF PENS AND THEY ARE ALL OVER THE UNITED STATES AND THREE OF THEM IN AUSTRALIA. THIS IS A PROJECT THAT HAS BROUGHT INTO ITS MIXED A GREAT NUMBER OF INVESTIGATORS AND I'M DELIGHTED TO SEE THE WAY IN WHICH THIS HAS RECRUITED -- PARTICULARLY DELIGHTED TO SEE THE WAY IN WHICH ENGINEERS HAVE JOINED US. SO AS YOU CAN SEE SINCE THE BRAIN BEGAN FY2014 IT IS WHEN IT START WE NOW HAVE GIVEN OUT 345 AWARDS SUPPORTING INVESTIGATORS A TOTAL INVESTMENT ALREADY OVER A HALF BECOME DOLLARS AND IT'S GOING TO CONTINUE. HAVE WE GOTTEN RESULTS? YEAH. ALL KINDS OF EXCITING THINGS ARE EMERGING. THIS IS JUST A SELECTION OF SIX PAPERS PUBLISHED IN 2017 AND THIS IS A SMALL REPUTATION. OVER 330 PUBLICATIONS TO DATE AND WE'RE STILL VERY MUCH ON THE RISING CURVE OF THAT. I COULD NAME A COUPLE OF THINGS THAT HAVE COME OUT OF THIS. IT WILL BE A LITTLE ARBITRARY BUT SOME THAT HAVE ATTRACTED PARTICULAR ATTENTION -- THE BRAIN INITIATIVE CELL CENSUS NETWORK LAUNCHED LAST YEAR A $250 MILLION EFFORT TO CATALOG THE PARTS LIST OF THE BRAIN IN MOUSE, IN MONKEY AND HUMANS WITH THE TECHNOLOGY NOW BECOMING POSSIBLE TO DO THIS. OBVIOUSLY A SOURCE OF GREAT POTENTIAL HERE AND GREAT EXCITEMENT AND WE ALSO HAVE TECHNOLOGIES THAT ARE INCREASINGLY DEVELOPING TO TRY TO DO WHAT I WAS TALKING ABOUT. TO BRIDGE THE SCALES BETWEEN THE BRAIN AND THE NEURON. I'M GOING TO STEAL A SLIDE IN TERMS OF THE USE OF ONE PHOTON OR TWO PHOTON IMAGING AT VARIOUS LEVELS. YOU CAN SEE THE CREDIT DOWN IN THE LOWER LEFT. YOU'RE LOOKING AT A MOUSE WHICH IS ACTUALLY RUNNING AND YOU'RE LOOKING TO SEE WHAT IS HAPPENING USING CALCIUM IMAGING AND YOU CAN LOOK ACROSS THE COURSE OF THE CORTEX AND THEN GO TO A MUCH HIGHER RESOLUTION AND LOO AT INDIVIDUAL CELLS. ALL OF THIS IN A MOUSE THAT IS UNDERGOING AN ACTIVITY IN REALTIME. PRETTY POWERFUL TO BE ABLE TO GET TO THIS LEVEL OF SENSITIVITY AND SPECIFICITY. AND WE ALSO WANT TO SEE HOW CAN WE BRING THESE OBSERVATIONS TO PATIENTS RECOGNIZING THAT THE BRAIN INITIATIVE -- WE WANT TO GRAB ON TO THOSE THERAPEUTIC PROMISE IN THIS CASE AND ALSO SOMETHING THAT WAS DISCUSSED YESTERDAY AFTERNOON BY OTHER COLLEAGUES AT THE UNIVERSITY OF FLORIDA COMING ONE WAYS TO USE WHAT WE'RE LEARNING ABOUT CIRCUITS TO MAKE DEEP BRAIN STIMULATION AND OTHER MODULATIONS EVEN MORE EFFECTIVE AND IN THIS INSTANCE I'LL PLAY A VIDEO FOR YOU -- SEVERELY AFFECTING THE LEFT ARM AND THIS IS A SITUATION WHERE THE STIMULATOR IS ACTUALLY TURNED ON BY THE INITIATION OF MOTOR ACTIVITY EVEN BEFORE THE INDIVIDUAL CLEARLY HAS STARTED THAT. THAT WAS THE STIMULATOR OFF AND NOW IT'S ON AND LOOK AT THE DIFFERENCE IN TERMS OF INCREDIBLELY SMOOTH. SO BRAIN IS JUST GETTING STARTED. I MENTIONED THE STABILITY OF THE SUPPORT WHICH WE DON'T ALWAYS GET TO ENJOY FOR PROJECTS WHERE WE SOMETIMES WORRY FROM YEAR TO YEAR WHAT IS GOING TO HAPPEN BUT IN THIS CASE WE HAVE BECAUSE OF THE 21st CENTURY CURSE BILL A TRAJECTORY THAT TAKES US OUT TO 2026. MAYBE THIS DIAGRAM PUZZLES YOU A BIT. THOSE ARE REAL DOLLARS. THE BASE FUNDING WHICH IS REGULAR APPROACH RACIALS FROM THE CONGRESS AND ON TOP OF THAT WHICH STARTS UP IN 2017 AND A LITTLE BIT MORE IN 2018. WE DON'T KNOW WHAT THE BASE FUNDING WILL BE. BUT ON TOP OF THAT YOU CAN SEE THE ADDITIONAL 21st CENTURY CURSE MONEY IS SOMETHING THAT HAS BEEN AGREED TO AND PASSED AND SIGNED INTO LAW. YOU MIGHT WONDER WHY IT'S SO HERKY JERKY AND MAYBE THIS IS NOT THE IDEAL WAY TO FUND SOMETHING IS THAT BIG CHUNK OF MONEY IN 2023. THEY FELT THEY COULD NOT MAKE THESE ALLOCATIONS WITHOUT OFFSETS. I DON'T REALLY REMEMBER WHAT IT WAS ABOUT 2023 WHERE THEY FELT THEY COULD GRAB SOME FUNDS AND PUT IT INTO BRAIN BUT THEY DID SO THINK ABOUT 2023 IS GOING TO BE A GREAT YEAR IF YOU'VE GOT SOME GOOD IDEAS GET THEM CUED UP FOR THAT. ANY WAY, AGAIN WE'RE VERY FORTUNATE TO HAVE THIS KIND OF STABILITY. CALL THAT A LITTLE LISTENS STABILITY BUT FOR NIH THIS IS GREAT. I THINK WE HAVE THE FORTUNATE CIRCUMSTANCE OF A TERRIFIC INITIAL SCIENTIFIC VISION THAT THE WORKING GROUP OF MY ADVISORY COMMITTEE TO THE DIRECTOR PUT FORWARD BACK THERE IN JUNE 2014 AND THAT WAS NOT SOMETHING THEY MADE UP ON SATURDAY AFTERNOON. THEY DID A LOT OF WORK HELD OVER THE SUMMER OF 2013. THEY WORKED QUITE INTENSIVELY SETTING SOME DRAFTS TOGETHER. COLLECTIVELY PUTTING THIS INTO A DOCUMENT WHICH IS UP ON THERE ON THE WEB AND DID DIVIDE THEM INTO THE FIRST FIVE YEARS AND THE SECOND FIVE YEARS. IT HAS SEEMED US TO AND TO WALTER AND JOSH WHO HAVE BEEN CAPABLE LEADERS AND A BIG SHOUT OUT TO THEM FOR THE WAY IN WHICH THEY'VE KEPT THIS MOMENTUM GOING AND PUTTING IN THEIR OWN PERSONAL TIME DESPITE THE FACT THEY ARE SUPPOSED TO BE RUNNING INSTITUTES, THIS HAS MADE IT POSSIBLE TO STEP BACK AND SAY FOR THE SECOND FIVE YEARS SCIENCE HAS MOVED FORWARD AND JUST SAY WELL THAT'S IT. IT'S TIME TO LOOK AT BRAIN 2.0 AND THAT IS WHAT WE'RE NOW AIMING TO DO WITH ANOTHER WORKING GROUP LOOKING AT WHERE WE'VE BEEN AND WHERE WE MIGHT BE ABLE TO GO AND SEE WHAT WE COULD DO TO DEVELOP THAT NEW VERSION OF THIS VISIONARY AND NEXT SET OF STEPS. HERE IS WHAT THIS IS GOING TO LOOK LIKE. AS YOU CAN SEE FROM CHECK MARKS DONE A COUPLE OF THINGS. A CALL FOR NOMINATIONS AND HERE WE ARE RIGHT NOW APRIL 18th WE WILL TOMORROW MORNING HAVE THE FIRST PUBLIC TOWN HALL AND THAT WILL BE THE KICK OFF FOR THIS WORKING GROUP THAT IS LOOKING AT BRAIN 2.0. IN THE COURSE OF THIS SUMMER THERE WILL BE AN RFI A REQUEST FOR INFORMATION WHEN WE ASK FOR WHAT WE MIGHT WANT TO INCORPORATE. THERE WILL BE WORKSHOPS HELD AROUND THE COUNTRY IN THE COMING MONTHS AND THE SUMMER AND THE FALL. BY A YEAR FROM NOW THERE WILL BE A DRAFT OF THIS PROPOSED NEXT FIVE YEARS PLAN AVAILABLE FOR PUBLIC COMMENT. NO DOUBT WE'LL GET PUBLIC COMMENT. PEOPLE WILL FEEL STRONGLY ABOUT THIS AND THEN THE FINAL VERSION OF THAT STRATEGIC PLAN READY FOR SUMMER OF 2019 JUST IN TIME TO PUT FORWARD THE NEXT SET OF FUNDING OPPORTUNITY ANNOUNCEMENTS FOR FY20 WHICH IS THE PLACE FOR WHICH THE NEXT FIVE YEARS WILL OFFICIALLY BEGIN. ALL OF THAT WILL BE HAPPENING THAT WILL PROBABLY ENGAGE A FAIR NUMBER OF YOU IN THESE DISCUSSIONS AND WORKSHOPS AND WE HOPE YOU WILL BE AS CREATIVE AS POSSIBLE IN ADVISING US. WHO'S GOING TO BE DOING THE WORK? HAPPY TO TELL YOU THAT WE'VE GOTTEN AGREEMENT OF ALL OF THE PEOPLE OF WHOSE NAMES ARE ON THE SLIDE AND THEY WILL BE WORKING INTENSIVELY AS YOU CAN TELL OVER THE COURSE OF THE NEXT YEAR WITH ABLE CHAIRS CATHERINE DUE LOCK AND JOHN MONSEL AND WE'LL SUPPORT THEM THROUGH NIH'S STAFFING AS BEST WE CAN TO MAKE THIS THE KIND OF THING THAT IS FUN BECAUSE YOU'RE FOCUSED ON THE SCIENCE AND NOT TOO MUCH ON THE BUREAUCRACY AND THE MEETINGS AND WE'LL TRY TO PROVIDE ALL OF THAT STAFF SUPPORT. AGAIN TOMORROW 8:30 THE OPPORTUNITY TO START THIS PROCESS WITH THIS WORKING GROUP TO BEGIN TO SHAPE WHAT THIS BRAIN 2.0 MIGHT LOOK LIKE AND I'M GOING TO BE VERY EXCITED TO SEE HOW THAT GOES FORWARD. SO THOSE ARE THE THINGS I WANTED TO PUT IN FRONT OF YOU IN A FEW BRIEF CONTINUES AS YOU ENTER THE -- SECOND DAY. THIS WORKING GROUP WILL BE PAYING ATTENTION TO ALL OF THE GREAT SCIENCE THAT IS BEING PRESENTED. A LOT OF POSTERS WHERE THE DETAILED STUFF IS BEING PRESENTED AND I HOPE EVERYONE TAKES TIME TO LOOK AT THIS THIS MORNING AND OTHER PARTS OF THIS MEETING. I WILL NOW TAKE ON THE NEXT STEP OF MY RESPONSIBILITY WHICH IS TO INTRODUCE YOUR KEYNOTE SPEAKER AND FAMILIAR TO MANY OF YOU TOM SUDHOF BORN IN GERMANY. HE WENT TO THE UNITED STATES TO DO A POST DOC AND THAT WAS NOT IN NEUROSCIENCE BUT WITH BROWN AND GOLDSTEIN WORKING ON THE RECENT FOR BUT WHEN HE GOT TO HIS OWN LABORATORY HE PIVOTED. HE IS ONE OF THE MOST DISTINGUISHED SCIENTIST. COGNITIVE STUFF IS IMPORTANT BUT LET'S FOCUS ON THE DETAIL. AS YOU KNOW HE RECEIVED THE 2013 NOBEL PRIZE IN PHYSIOLOGY AND MEDICINE FOR DISCOVERIES OF MACHINERY REGULATEING VEST CALL TRAFFIC. HOW SYNAPSE OCCUR IN THE BRAIN -- THAT FORM THE BASES FOR ALL INFORMATION PROCESSING BY THE BRAIN. HE IS HERE TO TALK ABOUT A TITLE OF TOWARDS MOLECULAR -- PLEASE JOIN ME IN WELCOMING TOM SUDHOF. [ APPLAUSE ] >> GOOD MORNING, EVERYBODY. THANK YOU VERY MUCH FOR THE TONIGHT TO TALK TO YOU AT THIS EARLY HOUR. I APPRECIATE YOU COMING ALTHOUGH I AM SURE IT WAS MUCH MORE INTERESTING ACTUALLY MUCH MORE ELEVATING TO HEAR IN MANY WAYS THE GOOD NEWS ABOUT NIH AND ABOUT THE WONDERFUL WORK THAT DR. COLLINS HAS BEEN DOING ON BEHALF OF ALL OF US. I CANNOT -- EMPHASIZE OVER AND OVER AGAIN HOW IMPORTANT IT IS TO HAVE A SUPPORTIVE FUNDING ENVIRONMENT AND YOU ALL KNOW THIS OF COURSE BUT I THINK IT HITS HOME IN PARTICULARLY IN STRONGLY IN THESE TIMES WHEN THERE SEEMS TO BE SO MUCH VOLATILITY IN GOVERNMENT AND IT'S REASSURING TO HEAR HOW THE FUNDING CAN AT LEAST HAVE SOME STABILITY IN THE FUTURE THAT WE CAN ALL COUNT ON FOR OUR WORK SO I WOULD REALLY LIKE TO THANK DR. COLLINS AND EVERYBODY ELSE FOR THEIR TIRELESS WORK ON OUR BEHALF. [ APPLAUSE ] SO NOW LET ME TALK ABOUT SCIENCE ABOUT THE DETAILS AS DR. COLLINS SO GRACIOUSLY INTRODUCED ME BUT BEFORE I DO THAT A DISCLOSURE. NOTHING I'M GOING TO SAY OR TALK ABOUT TODAY HAS ANYTHING TO DO WITH MY CONSULTING WORK. WHAT I'M GOING TO TALK ABOUT TODAY IS A PROJECT THAT IS VERY MUCH EVOLVING, EMERGING HOPEFULLY EMERGING CERTAINLY SOMETHING THAT IS UNFINISHED. AND BY WAY OF BACKGROUND AS YOU ALL KNOW UNDERSTANDING THE -- IS A HUGE CHALLENGE. FOR ME A BENCHMARK FOR THAT CHALLENGE IS THE FACT THAT THE BRAIN HAS A LOT OF NEURONS WHICH ARE CONNECTED IN TO VAST NETWORKS BY SYNAPSIS AND IF YOU COMPARE THE SIZE AND THE NUMBER INVOLVED HERE WITH THAT OF THE HUMAN GENOME IT IS DAUNTING BECAUSE WE ALL KNOW HOW A LITTLE WE UNDERSTAND THE HUMAN GENOME EVEN 15 YEARS AFTER ITS PUBLICATION AND IT MAKES IT CLEAR HOW MUCH MORE DIFFICULT IT WILL BE TRYING TO UNDERSTAND HOW THE BRAIN WORKS. THAT IS WHY WE'RE ALL HERE. WE ALL THINK THAT UNDERSTANDING THE BRAIN IS PROBABLY THE MOST INTERESTING AND MOST IMPORTANT CHALLENGE IN BIOMEDICAL RESEARCH. GIVEN ALL OF THE DISEASES THAT AFFECT THE BRAIN THAT WE CANNOT TREAT AND GIVEN THE FACT THAT WE ALL BELIEVE THAT THE BRAIN IN THE END THAT WHERE WE HAPPEN AND LIVE AS HUMANS -- HOW TO APPROACH THIS DIFFERS DRAMATICALLY. THIS IS A CLASSICAL VIEW OF THE -- ORGANIZATION OF THE BRAIN FIRST PUBLISHED BY -- WHICH SORT OF TRIES TO MAP DIFFERENT TYPES OF RESOLUTION YOU CAN HAVE IN THIS ANALYSIS AND I'VE ALWAYS BEEN BAFFLED BY THIS VIEW BECAUSE MY VIEW WOULD BE DIFFERENT. MY VIEW WOULD BE THIS VIEW HERE. AND YOU CAN SEE THAT THE ACTUAL SCALING OF THE SYSTEM IS COMPLETELY DIFFERENT BETWEEN THE SYSTEMS AND THE MOLECULAR VIEW BUT THERE IS A COMMON GROUND THAT UNITES THEM BUT THE IDEA THAT NEURONS ARE CONNECTED INTO CIRCUITS AND THIS IDEA I SUPPOSE IS THE COMMON GROUND THAT WE CAN ALL TALK -- TO EACH OTHER WHEN WE DISCUSS HOW THE BRAIN MIGHT WORK. WE ALL THINK I BELIEVE THAT THE WAY THE BRAIN POSSESS INFORMATION IS BY TRANSFERRING THAT INFORMATION IN CIRCUIT. CHANGING THE INFORMATION IN CIRCUITS THAT IN TURN ARE FORMED BY NEURONS WHICH ARE CONNECTED BY SYNAPSIS. SO LET'S LOOK AT THIS MORE CLOSELY. YOU CAN SEE HERE A SCHEMATIC VIEW OF A MICRO CIRCUIT IF YOU LIKE -- ON BOTH THE DOWNSTREAM ANOINT NEURON THAT IS -- ABOVE THERE IS A MIKE GRAPH OF A SIN NAP -- SYNAPSE. THE WAY HOW POSITION IS SUPPOSED TO BE POSSESSED IN THESE TYPES OF CIRCUITS IS AS THE ACTUAL POTENTIAL TRAVELS DOWN. THE FIRST AX OM ON THE LEFT THEY THEN FORM A BALL TO THE INHIBITORY NEURON SIGNALING. EVEN A SIMPLE CIRCUIT SUCH AS THAT CANNOT BE EASILY MODELED BECAUSE WHAT IS MOST IMPORTANT HERE IS NOT WHICH NEURON CONNECTS TO WHAT OTHER NEURON BY SYNAPSIS BUT WHAT THE PROPERTIES OF THESE SYNAPSIS ARE. -- THEY ALL OPERATE BY THE SAME PRINCIPLE IN USING NEUROTRANSMITTERS TO TRANSFER INFORMATION BUT THEY HAVE OTHER WAYS THAT -- EXTREMELY DIFFERENT PROPERTIES. LET ME ILLUSTRATE. IF YOU IMAGINE THAT THIS FIRST SYNAPSIS IS FACILITATING AND THIS IS DEPRESSING AND YOU HAVE STRAINS -- THEN ONLY HIGH FREQUENCY SIGNALS, SUBMITTED. SO YOU CANNOT CALCULATE WITHOUT UNDERSTANDING AND ALSO OTHER PROPERTIES WITHOUT UNDERSTAND THIS YOU CANNOT REALLY CALCULATE HOW A CIRCUIT MIGHT WORK. THIS IS THE PROBLEM THAT HAS INTERESTED ME IN RECENT YEARS WHICH IS HOW ARE SYNAPSIS FORMED. HOW IS IT THAT NEURONS ARE CONNECTED TO EACH OTHER AND HOW IS IT THAT THESE SYNAPSIS THEN ASSUME SPECIFIC PROPERTIES THAT ARE OFTEN -- IN A DEPENDENT MANNER. THIS PROBLEM WHICH I REFER TO IN A VERY PROVIDED SENSE INVOLVES NOT ONLY THE ACTUAL INITIAL FORMATION OF THE CONTACT BUT ALSO THE ASSEMBLY OF THE -- COMPONENT -- OF THOSE SYNAPSIS AS WELL AS THE CONFERRING OF SPECIFIC PROPERTIES ON TO THESE SYNAPSIS. WHAT SEE HERE IN THIS SCHEMATIC DIAGRAM ILLUSTRATED IS THE IDEA THAT THE GENERATION OF NEURONS AND MOST OF THE AX ONS ARE HAPPENING PRIMARILY DURING DEVELOPMENT. IT'S BALANCED BY THE ELIMINATION AND CAUSES A CONTINUOUS RESTRUCTURING OF NETWORKS OF CIRCUITS. IN THESE SYNAPSE FORMATION PROCESSES I AM NOT ONLY TALKING ABOUT THE GENERATION OF THE ACTUAL SYNAPSIS BUT ALSO RESTRUCTURING CHANGE IN THE PROPERTIES WHICH IS A SUBJECT THAT I'LL DISCUSS A LITTLE BIT MORE TODAY. SO THIS IS REALLY WHAT I AM INTERESTED IN TRYING TO UNDERSTAND AND THE IDEA HERE IS THAT THERE MUST BE A MOLECULE OF LOGIC THAT UNDER LICE ALL OF THESE PROCESSES. THERE MUST BE A WAY OF HOW THESE PROCESS ARE ENCODED IN THE GENOME. SYNOPSES ARE INTEREST CELLULAR JUNCTIONS. SIGH CITY. AND WE NEED TO LOOK AT THIS AS A PROBLEM OF CELL BIOLOGY. THESE MOLECULES THAT ARE INVOLVED ARE BASICALLY CLASSICAL RECEPTORS THAT ARE ACTIVATED AND STIMULATED AS -- THAT THEN ORGANIZE THE JUNCTIONS DEPENDING ON THE NEED THAT IS PREVALENT AT ANY GIVEN TIME. MANY MONITORS THAT ARE LIKELY TO ACCESS SYNOPSES HAVE BEEN QUIET. WHAT YOU SEE HERE IS AN OVERVIEW OF SOME OF THE MOLECULES ILLUSTRATING THE PRECITY NOTCHIC SIDE ON THE LEFT AND THE POST CITY NOTCHIC SLIDE ON THE LEFT. -- HOWEVER, FOR MOST OF THESE WE DON'T ACTUALLY REALLY KNOW WHAT EXACTLY THEY DO. IN FACT, THERE IS PROBABLY MORE TO BE DISCOVERED AND THE GOAL IS TO TRY TO UNDERSTAND WHAT SIGNALING PROCESSES EXACTLY MEDIATE WHAT PARTS OF THE SYNAPSIS -- EVENT. WE DON'T KNOW WHICH OF THESE ARE ONLY BYSTANDERS. I WANT TO EXPLAIN TO YOU THAT ONE PARTICULAR GROUP OF MOLECULES ARE CENTRAL ORGANIZERS OF SYNOPSES IN MANY DIFFERENT WAYS. THIS IS ALREADY MAYBE INDICATED IN THE SLIDE BY THE FACT THAT THEY BIND TO SO MANY AS ILLUSTRATED HERE BUT I'LL SHOW YOU THESE -- TO INSTRUCT THE POST SYNAPTIC SLIDE. SO MY TALK WILL FIRST INTRODUCE TO YOU -- AS A FAMILY AND GIVE YOU SOME STORIES ABOUT WHAT THEY MIGHT BE DOING IN DIFFERENT CONTEXT AND I WILL THEN FOCUS ON REALLY ONE PARTICULAR PROJECT WHICH REFERS TO THE REGULATION OF NEW AXON FUNCTION -- ONE PARTICULAR SITE. THE OVER-ALL GOAL I HAVE WITH MY TALK TODAY IS TO ILLUSTRATE HOW SYNAPTIC INFORMATION TRANSFER CAN BE CONTROLLED BY TRANCE SYNAPTIC SIGNALING. SO THIS IS REALLY WHAT I'M TRYING TO COMMUNICATE TO YOU IN ONE PARTICULAR EXAMPLE BUT BEFORE I GO ON WITH THIS LET ME INTRODUCE YOU. SO WAY SEE HERE IN THIS SLIDE IS AN OVERVIEW OF THE STRUCTURE OF NEW AXONS. THERE ARE THREE GENES. EACH COATS AN ALBA AND BETA -- THAT ARE SPECIFIED BY SPECIFIC PROMOTORS IN THE SAME GENE. THEY HAVE A COMPLEX -- AND THREE -- REPEATS. THEY HAVE ONLY A SINGLE AND NO EGIF REPEAT. ONE OF THE GENES HAS A GAMMA -- BUT ONLY THE SO-CALLED LOOP DOMAIN AS THE SINGLE -- DOMAIN. THESE THREE GENES ARE DIFFERENTIALLY PRESSED IN THE BRAIN AND IN PART LOCALIZED TO SYNOPSES. OKAY. OKAY. NO MORE INTERFERENCE. SO, THEY ARE AT LEAST IN PART LOCALIZED TO SYNOPSES. USING A MOUSE IN WHICH WE KNOCKED IN AN HA -- AND YOU CAN SEE THAT IT IS CLOSELY LOCALIZED WITH THE PRESYNAPTIC MARKER AND A JASE APARTMENT TO -- INSTEAD AND IN THE RECENT COLLABORATION WE SURPRISED ARINGLY OBSERVED THAT AX COMES ARE SPECIFICALLY LOCALIZED. EITHER -- OR IN SECTIONS ON THE BOTTOM AND YOU CAN SEE THAT IN EACH CASE THERE IS A LITTLE BLOB OF NEW AXOM ONE IN THE SYNAPTIC JUNCTION THAT IS RESTRICTED TO ONE PARTICULAR SUB DOMAIN. THESE SUB DOMAINS HAVE SOME INTERESTING PROPERTIES. THEY SEEM TO BE AROUND FIVE MOLECULES IN EACH CLUSTER. THIS CLUSTERS ARE OBSERVED EQUALLY. THE SIZE OF THESE CLUSTERS ARE OCCUPY ABOUT 10%, TO 20% OF THE TOTAL SIZE OF THE SYNAPTIC JUNCTION AS YOU CAN SEE HERE AND THEY CHANGE AS A FUNCTION OF -- BOTH IN -- THEY'VE MOVED TO A MORE CENTRAL POSITION SHOWN TO A MORE PERIPHERAL POSITION BUT STILL WITHIN THE SYNAPTIC -- SITUATION. THEY FORM A SUB DOMAIN. FOR EXAMPLE IN THAT THERE SEEMS TO BE ONLY AT THE LEVEL OF RESOLUTION ONLY A SINGLE SUB DOMAIN. THIS SUGGEST THAT EVEN THOUGH THEY LOOK HOMOGENEOUS ARE ACTUALLY MORE HETEROGENEOUS THAN WE ENVISIONED. THEY ARE SOMETIMES MUTATED IN REAL PSYCHIATRIC DISORDERS. HUNDREDS OF MUTATIONS OBSERVED IN PARTICULARLY AXOM 1 -- WHICH INCLUDE AMONG OTHERS SCHIZOPHRENIA, AUTISM, TERRET SYNDROME. IN FACT AS FAR AS I KNOW AXOM 1 IS STILL THE MOST COMMON GENE THAT IS MUTATED IN SCHIZOPHRENIA. IT TURNS OUT THAT THEY ARE ALL COPY NUMBER VARIATIONS. AXOM 1 IS SUCH A HUGE GENE THAT EVEN BIGGER CMVs COVER THE GENE. MOSTLY ASSOCIATED WITH MULTIPLE DISORDERS. NO SPECIFICITY. IT CAN BE ASSOCIATED WITH SIBLINGS. SUGGESTING THAT THERE IS NO DIRECT CAUSE EFFECT RELATIONSHIP BUT AN IN DIRECT PREDICTS POSITION CONFERRED. WE WONDER WHY THIS MIGHT BE. WE WERE PARTICULARLY DRIVEN BY THE IDEA THAT THERE MUST BE SOME TYPE OF GENETIC BACKGROUND EFFECT GIVEN THE FACT THAT THERE ARE SO MANY DIFFERENT TYPE OF CLINICAL MANIFESTS SO WE GENERATED NUANCE FROM HUMAN CELLS THAT CONTAIN A CONDITIONAL DELETION OF ONE -- TO TEST WHETHER UNDER A PRECISELY CONTROLLED GENETIC BACKGROUND THAT THERE WOULD BE A BIOLOGICAL EFFECT OF MUTATION IN AXOM 1. WHEN WE CULTURED THIS AS SHOWN HERE THEY ARE ABSOLUTELY IDENTICAL EXCEPT FOR -- WE FOUND THAT THEY FORM NORMAL SYNOPSES AND THEY LOOK PERFECTLY NORMAL BUT THEY EXPRESSED NOT UNEXPECTEDLY LESS -- AS SHOWN AT THE BOTTOM. WE ANALYZED THE SYNAPTIC FUNCTION AND FOUND THAT INDEED THERE WAS A DECREASE IN THE RESPONSIBLE SUSTAIN YOUR FREQUENCY THAT WAS PRESENT WAS THE MANY -- WAS UNCHANGED. THIS ALSO CAUSED THE DECREASE IN THE EVOKED AMPLITUDE OF EPSCs SUGGESTING THAT THERE IS A LOSS OF SYNAPTIC STRENGTH IN THESE NEURONS FINDING THAT WE'RE ALSO INVESTIGATING FURTHER USING CELLS FROM HUMAN PATIENTS AND IT LOOKS LIKE AND WE CAN CONFIRM IN THESE IPS CELLS. SO THESE EXPERIMENTS DEMONSTRATE THAT THIS IS DOING SOMETHING. THEY DON'T SOLVE THE PROBLEM OF TELLING US WHAT EXACTLY THE -- MUTATION IS DOING TO CAUSE IN THE END DISEASE IN A PATIENT BUT THEY ARE THE FIRST STEP TOWARD VALIDATING A GENETIC POPULATION IN A FUNCTIONAL MANNER. LET ME MOVE ON AND TELL YOU ABOUT A FEATURE THAT FORMS A CENTRAL THEME OF WHAT I'M GOING TO TALK ABOUT TODAY IS WHICH THE ENORMOUS SURPRISE -- SPLICING. LET ME HIGHLIGHT THESE SITES. TWO OF THEM ARE SHARED -- AND THEY ARE HIGHLY DIFFERENTIALLY EXPRESSED. THEY INDEED CREATE THOUSANDS OF -- AS CONFIRMED BY BIOSEQUENCING. THEY ARE HIGHLY -- BECAUSE OF THIS EXTREME DIVERSITY. -- THAT THEY ARE DIFFERENTIALLY DISTRIBUTED IN THE BRAIN AND WHEN WE USE SINGLE CELL TO TEST WHETHER THERE MIGHT BE A SPLICE CODE WE FOUND THAT THERE WAS A STRONG CORRELATION BETWEEN PARTICULAR COMBINATIONS OF SUPPLIES SITES -- AND MULTIPLE SPLICE VARIANTS AT THE SAME TIME AND THIS HAS GIVEN A NEW PATTERN ALTERNATIVE TO SPLICING BY AND LARGE COULD BE ASSOCIATED WITH A PARTICULAR TYPE OF NEURONS. HOWEVER, THERE WERE SOME SURPRISING FINDINGS AND LET ME ILLUSTRATE THIS TO YOU THAT FOCUSES ON SPLICE SIDE NUMBER FOUR. SO THIS PARTICULAR SPLICE SLIDE AS SHOWN HERE ON THE TOP INCLUDES OR EXCLUDES ONE -- BECK MEASURE THIS BY RPCR AND CALCULATOR INDEX THAT TELLS US HOW MUCH IS IN. AS SHOWN HERE FOR TWO DIFFERENT TYPES OF NEURONS. ONE TYPE OF NEURON PREFRONTAL CORTEX THE OTHER ONE A -- NEURONS THAT PROJECT. AND THE QUESTION HERE WAS WERE THE NEURONS THAT PROJECT THE SAME NEURONS HAVE SIMILAR -- COMBINATIONS OR NOT. IN THESE EXPERIMENTS WHAT I'M GOING TO SHOW YOU HERE IS ONLY SPLICE SET NUMBER FOUR AND THE BOTTOM GRAPH IS A PLOT OF IN VERSUS OUT AND THE X OX US ONE AND THE Y AXIS FOR NUMBER THREE AND YOU CAN SEE IMMEDIATELY THE TWO DIFFERENT TYPES OF NEURONS THAT ARE ILLUSTRATED HERE HAVE VERY DIFFERENT DISTRIBUTIONS. WHAT STRUCK US IS THAT -- THEY ARE ALTERNATIVELY SPLICED -- OUT FOR THE BLACK PREFRONTAL CORTEX NEURONS. THIS INDICATES -- IS THE SAME. AS YOU WOULD HAVE EXPECTED FOR TWO VERY -- GENES BUT THE VAST MAJORITY OF NEURONS WERE -- [ INAUDIBLE ] BUT OUT FOR THE SAME AS ONE. THERE IS ALWAYS SOME -- THAT HAVE AN INSERT AND SOME THAT DON'T WHICH GREATLY INCREASESth AMOUNT OF INFORMATION THAT YOU CAN CONVEY. MORE OVER THEY ARE HIGHLY -- THAT FOR ALL PURPOSES LOOKS THE SAME. IT IS NOT COORDINATED. THERE IS NOTHING LIKE THE SORT OF MARTZER REGULATOR THAT REGULATES BOTH IN THE SAME WAYS AND FINALLY -- A TYPE OF CODE WITH SPECIFICITY THAT CAN DISTINGUISH BETWEEN DIFFERENT NEURONS. IN THE STUDY THAT WAS PERFORMED THAT WAS RECENTLY PUBLISHED IT WAS SHOWN THAT AT THIS -- PARTICULAR SITE IS HIGHLY ACTIVITY DEPENDENT. WE HAVE SIMILAR DATA BUT I WOULD LIKE TO SHOW THIS DATA THAT WAS PUBLISHED LAST YEAR BECAUSE THEY ARE SO BEAUTIFUL IN MANY BAYS AND WHAT SEE HERE IS THE RESULTS OF PARTICLE NEURONS THAT THE AUTHORS PRESENTED B-R WHERE THEY STIMULATED THESE NEURONS WITH POTASSIUM CHLORIDE AND YOU CAN SEE THERE WAS A DRAMATIC CHANGE IN SLIDE SET NUMBER FOUR. AS ILLUSTRATED HERE ON THE LEFT WHEREAS ON THE RIGHT THE CONTROL SHOWS NO CHANGE. THIS SUGGESTED THAT THE SPLICING AT THE SITE IS DEPENDENT. THEY WENT ON TO EXPRESS THE SAME PHENOMENA USING THE PARADIGM AS SHOWN ON THE LEFT AND -- [ INDISCERNIBLE ] ISOLATED NEURONS THAT WERE ACTIVATED AND SEPARATED THEM FROM IN ACTIVE NEURONS AS SHOWN ON THE LEFT BY CELL SORTING. THEY THEN MEASURED SLIDES INVERSIONS OUT IN THESE ENACTIVE AND ACTIVE NEURONS AS SHOWN ON THE RIGHT AND THERE WAS A STRONG ACTIVATION IN -- SO THESE DEMONSTRATED THAT THIS SPLICING IS ACTIVE AND IT CHANGES. TODAY'S TALK WILL EXPLORE THIS PARTICULAR EVENT AND SPECIFICALLY ASK THE QUESTION WHAT IS THE FUNCTIONAL MEANING OF THIS SPLICING AT THIS PARTICULAR SITE? HOWEVER, BEFORE I MOVE ON TO TELL YOU ABOUT THIS LET ME FINISH UP MY INTRODUCTION TO TELL YOU MORE ABOUT THE DIVERSE -- [ INDISCERNIBLE ] AS YOU CAN SEE HERE IN THIS OVERVIEW SLIDE THERE IS MANY SUCH ITEMS. I ALREADY ILLUSTRATED THAT TO YOU EARLIER BUT YOU CAN SEE HERE IN THE PARTICULAR SLIDE IS THAT MANY OF THE INTERACTIONS ARE IN FACT REGULATED BY ALTERNATIVE SPLICING. ARE SOME MEDIATED BY BOTH. WHAT IS STRIKING TO ME PERSONALLY IS THAT NONE OF THESE SHARE ANY SEQUENCE WHICH ARE IN COMMON. ALL OF THEM ARE COMPLETELY DIFFERENT. AND IF ONE ASSUMES AS I HOPE YOU WILL AGREE WITH ME -- THIS MEANS THAT THE TRANCE SYNAPTIC SIGNALS THAT ARE CONFERRED ARE VERY DIFFERENT DEPENDING ON THE INTERACTIONS THAT ARE TAKING PLACE AT ANY GIVEN TIME. THESE VARIOUS -- ARE IN DIFFERENT ABUNDANCE. [ INDISCERNIBLE ] AND WHAT WE CREATE IN THIS MANNER IS WHAT I REFER TO AS A DYNAMIC NETWORK. AT ANY GIVEN TIME THERE ARE MANY DIFFERENT TYPES AND COMPLEXES WHOSE EXACT NATURE WE CANNOT PREDICT BECAUSE WE DON'T ACTUALLY KNOW -- THE INTERACTIONS BUT ARE LIKELY PLASTIC BECAUSE THE -- [ INDISCERNIBLE ] IS ACTIVITY DEPENDENT. SO MUCH FOR MY INTRODUCTION. I HOPE YOU'LL REMEMBER ALL OF THIS WHEN YOU WALK OUT OF HERE. MAYBE I SHOULD GIVE A TEST OR SOMETHING. NOW WE'RE GOING TO TALK ABOUT ONE PARTICULAR PROJECT. AND I'M GOING TO TALK ABOUT THIS ONE PARTICULAR PROJECT NAMELY THE BIOLOGICAL SIGNIFICANCE OF -- SPLICE SLIDE NUMBER FOUR IN TWO SECTIONS AND THE OVER-ALL GOAL OF MY PRESENTATION HERE IS TO ILLUSTRATE A SIMPLE MECHANISM ALTHOUGH ONE THAT IS EMBEDDED IN THE COMPLEX OVER-ALL SYSTEM THAT REGULATES THE FEATURES THE POST SEE NAPIC -- AT LEAST ON THE HIPPOCAMPUS. THIS SIMPLE. [ INDISCERNIBLE ] SO LET ME START OFF AND I'M GOING TO FIRST TELL YOU ABOUT SPLICE SLIDE NUMBER FOUR AND THE WAY THAT WE APPROACH THIS IS TO USE A GENETIC APPROACH THAT ON VOIDS POTENTIAL OVER EXPRESION ARTIFACT THAT'S COULD OCCUR IN RESCUE EXPERIMENTS. TO AVOID THIS AND TO GENERATE A SITUATION WHERE WE COULD CONTROL -- THE ALTERNATIVES -- WE USE THE GENETIC APPROACH -- WE DID THIS FOR FIRST SLIDE 3. IT HAS AN UNUSUAL SPLICE ACCEPT OR SEQUENCE THAT IS SHOWN HERE IN THE BOX. THIS UNUSUAL SEQUENCE BASICALLY HAS TOO MANY As. WHAT WE DID IS WE CONVERTED THIS UNUSUAL SEQUENCE IN TO A CONICAL SEQUENCE AS SHOWN BEFORE. [ INDISCERNIBLE ] THE RED LETTERS ARE THE ONES THAT WE CHANGED BECAUSE THIS IS NOW A CONICAL -- SIGH CONSEQUENCE -- IN ADDITION TO THAT WE FLANK THE AXOM 20 WITH -- SO WE CAN SIZE IT. [ INDISCERNIBLE ] AS A RESULT WHEN WE HAVE NO PREOR. [ INDISCERNIBLE ] THAT IS IN ACTIVE WE HAVE AN. [ INDISCERNIBLE ] GENO TYPE IF WE -- [ INDISCERNIBLE ] EVERYTHING ELSE IN THESE GENES IN THE NEW AXOM GENE IS NORMAL. [ INDISCERNIBLE ] WHEN WE ANALYZE THE NEURONS AND COMPARE THEM WITH AN UNRELATED CONTROL -- THIS IS ILLUSTRATED HERE AND DEMONSTRATION THAT -- WERE DECREASED ABOUT 50% BY. [ INDISCERNIBLE ] BUT WAS NORMALIZED BY THE DECISION. [ INDISCERNIBLE ] THEY WERE NORMALIZED BECAUSE THE WIRETAP PRIMARILY CONTAINS. [ INDISCERNIBLE ] AND IN THE RESPONSES ON THE OTHER HAND WERE NORMAL AS WERE THE IPSCs. THIS WAS AN UNEXPECTED FENO TYPE BUT THIS IS CLASSICAL FOR THE POST SYNAP CALL CHANGE. -- [ INDISCERNIBLE ] FOR THE NUMBER OF RECEPTORS IN THE SYNOPSES WAS DECREASED BOTH FROM ONE AND TWO NOT SHOWN HERE IN THE PICTURES. IN A SIMILAR MANNER DEMONSTRATING THAT THEY WERE DECREASED ON THE SURFACE. AT THE SAME TIME WE FOUND THAT IN THE SAME NEURONS WAS UNCHANGED IMPLICATING AN INCREASE IN THE INTERCELLULAR AT THE EXPENSE -- [ INDISCERNIBLE ] SO THESE EXPERIMENTS SUGGESTED THAT -- [ INDISCERNIBLE ] CAUSES THE CHANGE IN THE CONTEMPT AND RAISES THE QUESTION IF THEY ARE PRESYNAPTIC. WE HAVE FOLLOWED PRESYNAPTIC MOLECULES -- ALSO PRESENT ON POST SYNAPTIC -- AND VERY IMPORTANT THERE. SO THE. [ INDISCERNIBLE ] USING A PREPARATION THAT ALLOWS US TO SELECTLY MANIPULATE -- AND THIS PREPARATION IS THE CONNECTION OF PARAMETERS -- TO -- WHICH IS AN EXCITEED PROJECTION -- FROM THE HIPPOCAMPUS. WE USED STEREO TACTIC INJECTIONS. [ INDISCERNIBLE ] BY ELECTRO PHYSIOLOGY. WHAT SEE HERE IS THAT ONLY THE CRA NEURONS ARE AFFECTED BY THE ARVs IN THESE EXPERIMENTS THAT ARE ILLUSTRATED. [ INDISCERNIBLE ] THAT REFLECTS EGFP WHICH IS FUSED IN THESE EXPERIMENTS. WE SPECIFICALLY ONLY MANIPULATE PRESYNAPTIC NEURONS AND TEST WHETHER A PRESYNAPTIC HAS AN EFFECT ON POST SYNAPTIC -- [ INDISCERNIBLE ] THESE DISTINCT FORMS -- THEY DO EXIST DIFFERENT ELECTRICAL PROPERTIES AS ARE SHOWN HERE AND THEY ARE REGULAR -- NEURONS THAT FIRE REGULARLY WHEN YOU INJECT -- [ INDISCERNIBLE ] IN ADDITION TO THIS ELECTRICAL DIFFERENCE THESE NEURONS HAVE DIFFERENT FORMS OF LPT. [ INDISCERNIBLE ] THIS TOP LACKS JUST ALLOWS US TO SPECIFICALLY ANALYZE PRESYNAPTIC NEURONS SO WHAT DID WE FIND? WHAT WE FOUND THAT IS WE COULD CONFIRM WHAT WE HAD SEEN. AND. [ INDISCERNIBLE ] AS SHOWN HERE BOTH FOR -- [ INDISCERNIBLE ] WERE DECREASED BY PRESYNAPTIC. [ INDISCERNIBLE ] BUT WERE NORMALIZED AS SHOWN HERE BY PRESYNAPTIC INCISION OF SPLICE SLIDE 4. [ INDISCERNIBLE ] THEY WERE NOT AFFECTED. THEY WERE EXACTLY THE SAME. WE ALSO MEASURED LPT AND WE EXPERIENCED SOMETHING OF A SURPRISE BECAUSE AS YOU REMEMBER IN THESE NEURONS THE RECEPTORS THEMSELVES ARE NOT CHANGED. IT'S JUST THAT THE DISTRIBUTION HAS CHANGED. THEY ARE SHIFTED TO THE INSIDE. [ INDISCERNIBLE ] LPT -- IT'S BEEN SHOWN BY CLASSIC WORK. [ INDISCERNIBLE ] THAT. [ INDISCERNIBLE ] OF AMPLE RECEPTORS TO POST SYNAPTIC SPECIAL LACKSES. THIS MEANS THAT IF THERE ARE MORE. [ INDISCERNIBLE ] THE POST SYNAPTIC -- LPT WAS NEARLY ABOLISHED. THIS COULD BE COMPLETELY REVERSED BY PRESYNAPTIC -- DEMONSTRATING THAT IT IS INDEED A -- CONTROLLED GATE. [ INDISCERNIBLE ] WAS NOT AFFECTED. THERE WAS NO CHANGE IN PRESYNAPTIC LPT IN. [ INDISCERNIBLE ] SO WHAT I SHOWED YOU THEN -- IS THAT. [ INDISCERNIBLE ] DICTATES THE POST SYNAPTIC PROPERTIES IN TERMS OF AMPLE RECEPTORS AND IN TERMS OF LONG-TERM -- [ INDISCERNIBLE ] THIS SPECIFIES THE VARIOUS SYNOPSES THAT I DISCUSSED WITH YOU. WHAT ABOUT AXOM ONE. I'LL DISCUSS WITH YOU WHAT HAPPENS AT THE SAME SPLICE SLIDE OF AXOM ONE AND THE TITLE OF THE TALK GIVES AWAY THE CONCLUSION. BUT WHEN WE STARTED THIS WE THOUGHT THAT THEY WERE ALL SIMILAR. [ INDISCERNIBLE ] THEY ARE 100% IDENTICAL. WE THOUGHT THESE WOULD BE DOING THE SAME THING AND I HAD A HARD TIME CONVINCING -- TO PURSUE THIS PROJECT. I HAD TO FIND SOMEBODY WHO WAS TOTALLY IN EXPERIENCED AND HAD NO IDEA TOGETHER TO DO THESE EXPERIMENTS BECAUSE IT SOUNDED KIND OF -- WRONG. THEY ALL LOOK THE SAME. THEY SHOULD BE DOING THE SAME. WE ARE USING THE SAME APPROACH THAT I MENTIONED BEFORE. IT TURNS OUT THAT AGAIN THE AXOM [ INDISCERNIBLE ] THAT IS NONCONICAL AGAIN WE MUTED IT. [ INDISCERNIBLE ] INTO A CONICAL ONE AS SHOWN HERE WITH THESE WHITE LETTERS AND THIS AGAIN CREATED A MOUSE IN WHICH. [ INDISCERNIBLE ] WAS IN NEVER OUT. HOWEVER. [ INDISCERNIBLE ] WE COULD CONVERT THIS AGAIN TO AN -- FENOTYPE. -- AND WHAT WE FOUND WAS THAT AMPLE. [ INDISCERNIBLE ] IN THE. [ INDISCERNIBLE ] PATHWAY WERE AGAIN NOT CHANGED AT ALL. THEY WERE EXACTLY THE SAME. THERE WAS NO DECREASE IN THE RESPONSES AS SHOWN HERE IN. [ INDISCERNIBLE ] OR IN REGULAR. [ INDISCERNIBLE ] NEURONS. SO RECEPTORS WERE NOT AFFECTED BUT. [ INDISCERNIBLE ] WE FOUND THAT IF THERE WAS A PRESYNAPTIC AS FOUR PLUS THERE WAS AN INCREASE IN THE RECEPTOR RESPONSES THAT WAS QUITE MASSIVE AND THAT WAS COMPLETELY REVERSE WHEN WE PRESYNOPTICALLY SLIDE SET NUMBER 4. YOU'VE DIFFERENT PHENOTYPE. THAT IS CONTROLLED BY SLIDE SET NUMBER 4. VERY DIFFERENT FROM SLIDE SET 3. SUGGESTING THAT THE SAME IN THE SAME SYNAPSIS DOES COMPLETELY DIFFERENT THINGS IN TERMS OF THE POST SYNAPTIC TARGET BUT NOT IN WHAT THEY DO. IT'S INTERESTING IN THIS CONTEXT THAT. [ INDISCERNIBLE ] MANY PEOPLE, MANY PSYCHIATRISTS BELIEVE THAT THAT THE RECEPTORS HAVE SOMETHING TO DO WITH SCHIZOPHRENIA. I DON'T KNOW HOW TRUE THAT IS. BUT ANY WAY -- WE MEASURED LPT AND WHAT WE FOUND MAYBE NOT SURPRISINGLY IS NOW. [ INDISCERNIBLE ] IT ACTUALLY ENHANCED LPT AS SHOWN HERE. THIS ENHANCEMENT MAY NOT BE SURPRISING BECAUSE YOU HAVE BETTER INDUCTION SO YOU CAN EASILY EXPLAIN IT BUT IT ILLUSTRATES THE FACT THAT THESE TWO DIFFERENT MANIPULATIONS HAVE OPPOSITE EFFECTS. SO THE OVER-ALL ASSESSMENT IS. [ INDISCERNIBLE ] IN ONE PARTICULAR. [ INDISCERNIBLE ] CAUSED OPPOSITE EFFECTS. [ INDISCERNIBLE ] AND OPPOSITE EFFECTS AS SHOWN HERE ON LONG-TERM. [ INDISCERNIBLE ] [ INDISCERNIBLE ] I DIDN'T DISCUSS THEM BECAUSE I DIDN'T THINK THEY WOULD BE THAT INTERESTING BECAUSE I DIDN'T HAVE ANY PHENOTYPE. [ INDISCERNIBLE ] DETERMINES THE POST SYNAPTIC RECEPTOR COMPOSITION OVER A LARGE RANGE AND REMEMBER I TOLD YOU EARLIER. [ INDISCERNIBLE ] IT CAN BE INDEPENDENTLY REGULATED AND ITS ACTIVITY DEPENDENT SO IT CAN CHANGE AS A FUNCTION OF ACTIVITY ALTHOUGH. [ INDISCERNIBLE ] HAVE NOT BEEN EXPLORED. THIS TYPE OF MECHANISM IT IS RELATIVELY SIMPLE MECHANISM BY WHICH A PRESYNAPTIC NEURON CAN IN EXPECT THE POST SYNAPTIC NEURON IN TERMS OF THE RESPONSE TO A GIVEN NEUROTRANSMITTER SIGNAL. NOW MANY PEOPLE BELIEVE THAT -- [ INDISCERNIBLE ] RECEPTORS ARE SOMETHING OF A SIDEKICK. [ INDISCERNIBLE ] AND I THINK THIS IS A MISCONCEPTION AS PROBABLY -- BEEN DOCUMENTED AND. [ INDISCERNIBLE ] CONTINUOUSLY CONTRIBUTE VIRTUALLY ALL TRANSMISSION AND ARE ESSENTIAL IN MOST CIRCUMSTANCES. IF THE OVER-ALL CONCEPT IS TRUE THE QUESTION THEN ARISES WHETHER OVER AN EXPRESSION OF. [ INDISCERNIBLE ] WOULD DIRECTLY EFFECT THE RECEPTOR. [ INDISCERNIBLE ] IN MICE THAT HAVE NOT BEEN CHANGED SO WE ALSO TESTED THIS USING THE SAME PARADIGM THAT I ALREADY DEMONSTRATED MULTIPLE TIMES. [ INDISCERNIBLE ] ANALYSIS TWO WEEKS LATER AND IN THIS CASE WE. [ INDISCERNIBLE ] DIFFERENT FORMS OF. [ INDISCERNIBLE ] THAT ONLY DIFFER IN THE SPLICING. [ INDISCERNIBLE ] THEY ARE MUCH SHORTER. WHAT SEE HERE IS THAT INDEED WE SEE THE EXPECTED EFFECT. THIS IS FOR MINUS AND THIS IS FOR PLUS AS SHOWN HERE. [ INDISCERNIBLE ] WE CAN AMPLIFY INCREASE THE. [ INDISCERNIBLE ] RESPONSE. [ INDISCERNIBLE ] IF IT'S FOR MINUS NOTHING HAPPENS. CONVERSELY WHEN WE OVER -- [ INDISCERNIBLE ] THE. [ INDISCERNIBLE ] SUPPRESSES RESPONSES AS HOPE HERE SO THIS SYSTEM WORKS NOT ONLY. [ INDISCERNIBLE ] IT ALSO SHOWS THAT OVER EXPRESSION CAN CAUSE THE SAME EFFECTS IN THIS PARADIGM DEMONSTRATING THAT THERE IS A TRANCE SYNAPTIC CONTROL OF THE AMPLE RECEPTOR RESPONSES SO WHAT I'VE TRIED TO TELL YOU TODAY IS THAT NO AXOMS IS REGULATED. [ INDISCERNIBLE ] JUST REMEMBER SPLICE SLIDE NUMBER 4 AND THAT 1 AND 3 HAVE OPPOSITE EFFECTS AND THE LAST FEW SLIDES I WOULD LIKE TO ARGUE THAT THIS IS POTENTIALLY IMPORTANT IN UNDERSTANDING HOW CIRCUITS OPERATE AND I WOULD LIKE TO GIVE YOU A GLIMPSE OF WHAT WE'RE THINKING. SO AS YOU REMEMBER SYNAPSIS OPERATE -- [ INDISCERNIBLE ] IN NEURO CIRCUS. CIRCUITS. IT INVOLVE MANY DIFFERENT TYPES OF SIGNALS AND CAN HAVE OTHER RESPONSES AS WELL. BY -- CONTROLLING THE. [ INDISCERNIBLE ] ALTERNATIVE SPLICING OF 1 AND 3 AND SLIDE SET NUMBER 4 CAN PRO FOUNDLY EFFECT HOW -- [ INDISCERNIBLE ] OCCURS IN GIVEN SYNAPSIS AND IN UNDERSTANDING HOW THIS MIGHT WORK HOW THIS MIGHT BE CONTROLLED WITH THE MOLECULAR MECHANISMS -- WILL BE NECESSARY IN ORDER TO UNDERSTAND IN ORDER TO PREDICT HOW CIRCUITS OPERATE. IN TERMS OF THE OVER-ALL FUNCTION OF. [ INDISCERNIBLE ] WHAT IS OBVIOUS WHEN YOU LOO AT. [ INDISCERNIBLE ] AND THEIR VARIOUS INTERACTIONS WITH DIFFERENT. [ INDISCERNIBLE ] IT IS THAT THEY LOOK LIKE PLATFORMS. PLATFORMS THAT INTERACT WITH MANY DIFFERENT OTHER MOLECULES AND THERE BY -- DISTINCT SIGNALS. THIS IS JUST ONE LITTLE BIT OF THE. [ INDISCERNIBLE ] AT LEAST WHICH HAVE MANY MORE DOMAINS. AS SIGNALING PLATFORMS WE EXPECT THAT THEY WILL BE INVOLVED IN OTHER PROCESSES AS WELL THAT I HAVE NOT TALKED ABOUT TODAY AND THE WAY THAT WE ENVISION THIS IS WITH THIS ILLUSTRATION THIS MODEL IMAGINE THIS IS -- IT HAS MANY BELLS AND WHISTLES -- AND WE ENVISION THAT ITS DIFFERENT FEATURES MAY MEET AT DIFFERENT TYPES OF INTERACTIONS. I ONLY DISCUSSED -- AMBULATORY RECEPTORS TODAY. [ INDISCERNIBLE ] WE KNOW THEY HAVE SOMETHING TO DO WITH. [ INDISCERNIBLE ] RECEPTORS. [ INDISCERNIBLE ] AND AS SUCH WE BELIEVE THAT VARIOUS -- [ INDISCERNIBLE ] MAY SPECIFY SYNAPSIS PROPERTIES. [ INDISCERNIBLE ] IN THIS DYNAMIC INTERACTION NETWORK THAT I INTRODUCED TO YOU EARLIER ON. [ INDISCERNIBLE ] THE PROPERTIES OF SYNOPSES AND THUS WE THINK THAT THE. [ INDISCERNIBLE ] FUNCTION IN SPECIFICATION AND THAT OTHER MOLECULES. [ INDISCERNIBLE ] ARE THE. [ INDISCERNIBLE ] FEATURES. LET ME CLOSE BY ACKNOWLEDGING THE MAJOR -- CONTRIBUTORS TO THE WORK THAT I DISCUSSED WITH YOU TODAY. [ INDISCERNIBLE ] SUPPORTED BY. [ INDISCERNIBLE ] AND JASON -- [ INDISCERNIBLE ] DID THE AND INITIAL WORK. [ INDISCERNIBLE ] THANK YOU VERY MUCH FOR YOUR ATTENTION. [ APPLAUSE ] AND JUST IN CLOSING LET ME SPECIFICALLY SAY I'M REALLY GRATEFUL FOR THE MONEY. QUESTIONS? HAPPY TO ANSWER ANY QUESTIONS. >> WHAT TIME SCALE DO THESE CHANGES -- EXPRESSION COMPOSITION OCCUR. MINUTES, HOURS, DAYS? >> THEY SUGGESTED IT'S WITHIN HOURS. NOT MINUTES. NOT DAYS. >> IS THERE ANY CHANGE BETWEEN WAKING AND SLEEPING IN THE. [ INDISCERNIBLE ] >> THERE IS A BEAUTIFUL PAPER FROM ABOUT 10 YEARS AGO FROM -- WHO SHOWED THERE WAS A. [ INDISCERNIBLE ] IN THE BRAIN WHATEVER THAT MEANS. >> TOM, A BEAUTIFUL TALK. YOU SHOWED THAT YOUR -- IS LOCALIZED IN A SMALL DOMAIN IN COLLABORATION -- [ INDISCERNIBLE ] IT TURNS OUT THAT ANTIBODIES IS A CRUCIAL ISSUE IN ALL OF THESE STUDIES BECAUSE NONE OF THE AVAILABLE ANTIBODIES ARE ALL THAT GREAT SO WE DON'T KNOW WHETHER THAT HAPPENS WITH AXOM 3. [ INDISCERNIBLE ] I DON'T KNOW THAT. THAT IS A GOOD QUESTION. >> AND WITH YOUR SWISS ARMY KNIFE ANALOGY -- YOU THINK ORGANIZED THE WAY TO SLIDE THE KNIFE OR A LOT OF INTEREST MIXING. >> WHAT IS THE MECHANISM. [ INDISCERNIBLE ] WHAT WE REALLY NEED TO DO IS MEASURE REAL AFFINITIES THAT HAS NEVER BEEN DONE. IT COULD NOT BE PUBLISHED BECAUSE -- ARE NOT INTERESTED IN THIS KIND OF STUFF BUT THAT IS REALLY WHAT WE ACTUALLY NEED BECAUSE THERE IS PRELIMINARY DATA THAT THE INTERACTIONS MAY ACTUALLY BE QUITE DIFFERENT. THEY COULD DIFFER BY AS MUCH AS -- IN TERMS OF AFFINITIES BETWEEN DIFFERENT ICE FORMS -- AND THAT THIS ENORMOUS INTERACTION NETWORK THAT I TRIED TO INTRODUCE TO YOU MAY SHIFT AS A FUNCTION -- [ INDISCERNIBLE ] AND THIS IS HOW WE THINK IT WORKS. >> LIKE JOSH I WAS STRUCK BY THE LOCALIZATION. [ INDISCERNIBLE ] AT THE SYNAPSE INSTEAD OF THE BROAD SCALE -- [ INDISCERNIBLE ] OR WHETHER THE PRESYNAPTIC ADHESION COMPLEXES ARE SIMILARLY RESTRICTED. [ INDISCERNIBLE ] WE DIDN'T EXPECT THAT THEY WOULD BE DISTRIBUTED. >> NO IT'S NOT PREDICTED. THERE IS A BEAUTIFUL STUDY FROM TOM -- AND THOMAS WHICH SHOWS THE PERIPHERY THERE IS AN EARLIER WORK THAT. [ INDISCERNIBLE ] OUTSIDE THE JUNCTION. THERE ARE CLASSICAL STUDIES, BEAUTIFUL STUDIES FROM -- [ INDISCERNIBLE ] SHOWING. [ INDISCERNIBLE ] IN THE JUNCTION ITSELF AND THEY HAVE ADHESION TYPE DOMAINS. THAT IS ABOUT IT. >> SO SUGGEST THAT THESE ARE PROBABLY ONE OF MANY DIFFERENT COMPONENTS THAT ARE IMPORTANT FOR ADHESION BUT WHAT DO YOU THINK IS GOING ON IN THESE DOMAINS? SO OUR VIEW OF THIS IS THAT THE MOST LIKELY EXPLANATION OF THESE FINDINGS IS THAT THERE IS MORE STRUCTURE TO A SYNAPTIC JUNCTION THAN IS APPRECIATED FROM EM. THAT WE BELIEVE THAT ALL OF THESE ADHESION MARKERS PRIMARILY ACT AS SIGNALING MARKERS. WHAT THEY DO IS ACTUALLY SIGNAL AND THAT THEY ARE SIGNALING HUBS THAT THEY IN OTHER WORDS ALTERNATE SIGNALING IN ONE PARTICULAR SPACE. HOW THE MECHANISM OF THAT OPERATES I DON'T KNOW. >> MY QUESTION WAS A FOLLOW-UP OF THIS ONE WHICH IS HOW DO YOU THINK -- SIGNALING IS HAPPENING? [ INDISCERNIBLE ] THAT CAUSES AN ACTIVE INSERTION AND REMOVAL OF RECEPTORS. [ INDISCERNIBLE ] BY MANIPULATING THE. [ INDISCERNIBLE ] SITUATION. >> I DIDN'T GO INTO THIS BUT IT TURNS OUT THAT THE SIGNALING THAT CONTROLS POST SYNAPTIC RECEPTORS BY. [ INDISCERNIBLE ] DID-- DOES NOT REQUIRE. [ INDISCERNIBLE ] AS LONG AS THE SEQUENCES ARE ANCHORED BY A. [ INDISCERNIBLE ] ANCHOR IT WORKS. HOWEVER OTHER FUNCTIONS -- DO REQUIRE THE. [ INDISCERNIBLE ] SO WE AGAIN THINK THAT THERE IS MULTIPLE PATHWAYS THAT DEPEND ON DIFFERENT TYPES OF POST SYNAPTIC. [ INDISCERNIBLE ] THAT ARE RESPONSIBLE. WHAT IS THE MECHANISM IS I DON'T KNOW. >> YOU MENTIONED -- [ INDISCERNIBLE ] WAS CHANGED BUT THAT THE TOTAL NUMBER OF RECEPTORS -- COULD IT BE -- MOVEMENT, MOTION. [ INDISCERNIBLE ] >> GREAT TO SEE YOU. DIDN'T EXPECT YOU HERE. WHAT HE REFERS TO IS A DISCUSSION IN THE FIELD. [ INDISCERNIBLE ] BY OTHERS. [ INDISCERNIBLE ] THAT A.M. ARE BOTH IN SYNOPSES AND OUT OF SYNOPSES ON THE SURFACE AND THE QUESTION IS IT POSSIBLE THAT. [ INDISCERNIBLE ] THAT THEY MOVE AWAY. WE HAVE LOOKED APARTMENT THIS IN A PRELIMINARY MANNER AND FIND THAT MOST ARE INTERCELLULAR BUT THIS IS ACTUALLY A LITTLE MORE DIFFICULT THAN IT SOUNDS BECAUSE IF THEY ARE DIFFUSED ALL OVER YOU CANNOT LOCALIZE THEM. YOU NEED SOME KIND OF BETTER WAY OF LOOKING AT THAT. SO THE BEST EVIDENCE WE HAVE IS THAT PUFFS OF. [ INDISCERNIBLE ] SHOWED -- [ INDISCERNIBLE ] >> SIMPLE QUESTION. I JUST WONDER IF YOU HAVE. [ INDISCERNIBLE ] WITH THE TRANSMITTER. >> THE COORDINATION OF WHAT? >> THE TRANSMITTER WHICH MEANS IF YOU LOOK AT A. [ INDISCERNIBLE ] OF THE TRANSMITTER YOU USE A DIFFERENT TYPE OF TRANSMITTER. [ INDISCERNIBLE ] >> I TALKED ABOUT ONE TYPE OF. [ INDISCERNIBLE ] AND THERE ARE MANY DIFFERENT TYPES THAT ARE DIFFERENT. SO SYNOPSES ARE -- DIFFERENT. SO WHAT I TALKED ABOUT TODAY WAS. [ INDISCERNIBLE ] [ APPLAUSE ] IT'S MY PLEASURE TO INTRODUCE THE CLOSING ACT OF THE DAY IN ADDITION TO MY INTRODUCTION OF THE OPENING OF THE ACT EARLIER TODAY. SHE GREW UP IN LEBANON. WHERE SHE STARTED MEDICAL SCHOOL AT THE UNIVERSITY OF BEIRUT BEFORE TRANSFERRING DURING THE LEBANESE CIVIL WAR. SHE IS -- HAD AN INCREDIBLY ACCOMPLISHED CAREER SINCE AS YOU MAY KNOW. MOST NOTABLE FOR WHAT I THINK IS ONE OF THE TRULY COMPELLING TRANSLATIONAL STORIES THAT STARTS WITH A CLINICAL OBSERVATION CONTINUES WITH INTENSE CURIOSITY FOCUS AND DRIVE AND ENDS UP WITH A REMARKABLE DISCOVERY AND THAT IS HER DISCOVERY FOR THE GENE FOR THE RET SYNDROME. AND REALLY HAS PAVED THE WAY TOWARD A MOLECULAR UNDERSTANDING OF MANY OF THESE DISEASES THAT AFFECT THE NEURO SYSTEM. SHE IS AND INVESTIGATOR OF THE HOWARD HUGHES MEDICAL INSTITUTE AND A PROFESSOR OF PEDIATRIC'S MOLECULAR AND HUMAN GENETICS. A BAYLOR COLLEGE OF MEDICINE AND PLEASE WELCOME HER TO THE STAGE TO GIVE A TALK ON UNDERSTANDING THE BRAIN ONE PATIENT AND ONE ANIMAL MODEL AT A TIME. [ APPLAUSE ] >> THANK YOU. IT'S A PLEASURE TO BE HERE TODAY. I'M NOT FUNDED BY THE BRAIN INITIATIVE ALTHOUGH I'M GOING TO HAVE TO LEARN QUICKLY STARTING TOMORROW ABOUT IT. BUT I REALLY BENEFITED FROM A LOT OF WORK BY PEOPLE IN THIS ROOM SO I APPRECIATE THAT. I WOULD LIKE TO HAVE SOME DISCLOSURES. I DO GET COLLABORATIONS WITH UCB ON DEVELOPING PHARMACEUTICALS TO ENZYMES THAT WE'VE DISCOVERED. I COLLABORATE WITH -- ON THE USE OF -- FOR ONE DISEASE THAT I'M GOING TO TALK ABOUT TODAY. I DON'T GET MONEY FROM THEM BUT WE DO COLLABORATE. SO UNDERSTANDING THE BRAIN ONE PATIENT AT A TIME. THAT IS HOW I GOT INTO SCIENCE AND THAT IS HOW I GOT INTERESTED IN THE BRAIN. SEEING PATIENTS WITH VERY PERPLEXING NEUROLOGICAL DISORDERS AND THIS CONTINUES TO BE THE CASE. AND IN TODAY'S TALK WHAT I WOULD LIKE TO DO IS SHARE WITH YOU HOW I ACTUALLY TO THIS DAY CONTINUE TO LEARN. MY LAST MOST FASCINATING PATIENT I SAW TWO MONTHS AGO AND HE CAME BACK AND SAW ME SUNDAY NIGHT BECAUSE I WAS TRAVELING YESTERDAY. WE DO CONTINUE TO LEARN FROM PATIENTS AND I'LL SHARE WITH YOU HOW THE SUBTILTIES OF NEUROLOGICAL DYSFUNCTION OF PSYCHIATRIC DYSFUNCTION ARE SOMETIMES EASIER SEEN IN PATIENTS. BUT IN ADDITION I'VE BEEN FORTUNATE TO BENEFIT FROM MAKING ANIMAL MODELS. WE'VE DONE FLY WORK AND MOUSE WORK. WHERE THE MOUSE ACTUALLY TAUGHT ME SOMETHING ABOUT HUMAN CLINICAL DISORDER SO WE'LL DISCUSS THAT. AND I WILL BE TELLING YOU STORIES ABOUT TWO DIFFERENT CLASSES OF DISORDERS. BOTH AFFECT CHILDREN. AND THE REASON I CHOSE THESE TWO DISORDERS BECAUSE YOU'LL SEE THEY HAVE IMPLICATIONS WITH HOW WE NEED TO WORRY ABOUT PSYCHIATRIC DISORDERS. WE'VE MADE A LOT OF PROGRESS IN NEUROLOGICAL DISEASES. BUT PSYCHIATRIC DISEASES IS TOUGH AND LEARNING LESSONS I WOULD LIKE TO SHARE WITH YOU SOME THOUGHTS WITH HOW WE MOVE FORWARD IN THINKING ABOUT PSYCHIATRIC DISEASES. MY PASSION WAS DRIVEN BY THIS ONE PATIENT THAT I SAW HER WHEN I WAS AN INTERN. SHE LOST MANY SKILLS AND YOU SEE HER HERE WRINGING HER HAND AND SO MANY AFTER PUBLISHING A SMALL PAPER ABOUT THESE PATIENTS MANY CAME TO OUR CENTER AND HUNDREDS OF THEM -- THEY WERE ALL SPORADIC AND THIS WAS 1985 WHEN THE GENOME WAS NOT YET MAPPED. WE UNDERSTAND THE BRAIN BUT WE DIDN'T HAVE THE TOOLS BUT WE DID MANAGE TO FIND THE DISORDER IS A GENETIC DISEASE AND CAUSED BY A MUTATION. WHAT I WOULD LIKE TO SHARE WITH YOU IS WHAT WE LEARNED FROM THE PATIENTS. THIS IS ON THE X CHROMOSOME WHICH MEANS HALF OF THE CELLS WILL HAVE A HEALTHY -- AND THE OTHER HALF WILL HAVE MUTED -- HALF OF THE CELLS WILL NOT HAVE A FUNCTIONAL COPY AND THIS IS BECAUSE OF -- IN ACTIVATION AND YOU'LL HAVE THE CLASSIC RHETT SYNDROME. IN A FEMALE YOU'LL HAVE REGRESSION AND INTELLECTUAL DISABILITY -- MOTOR PROBLEMS AND INABILITY TO EXECUTE, SEIZURES AND EVENTUALLY PARK SOMISM. EVERY PART OF THE CENTRAL NERVE SYSTEM IS AFFECTED. I CANNOT LIST EVERYTHING HERE. IN MALES WHERE YOU ONLY HAVE ONE X CHROMOSOME. THE GENE IS MUTATED. IT'S QUITE SEVERE. YOU CAN HAVE AN INFANT THAT IS NOT GOING TO MAKE IT. THEY WILL NEED BREATHING SUPPORT. MOTOR PROBLEMS AND THEY WILL DIE TYPICALLY IN THE FIRST TWO YEARS OF LIFE. IF THEY HAVE PARTIALLY DYSFUNCTIONAL BUT STILL SEVERE MUTATION THEY MAY DIE AT 10-YEAR-OLD OR 15-YEAR-OLD. BUT THE LESSONS CAN BE LEARNED WITH THOSE WITH MILDER MUTATIONS AND THIS IS WHAT I WANT TO TAKE A MEANT OR TWO TO TELL YOU ABOUT. THERE ARE HYPER MORE FICK MUTATIONS I MEAN EMU TAKES THAT PARTIALLY IN ACTIVELY -- THE PROTEIN. THIS MALE MIGHT PRESENT IN DIFFERENT COLORS BECAUSE THEY ARE DIFFERENT PRESENTATIONS. DEPENDING ON THE MUTATION. THEY WILL ALWAYS HAVE MILD LEARNING DISABILITY. BUT MAY PRESENT WITH AUTISM. ANXIETY AND OBSESSIVE COMPULSIVE DISORDER. QUITE A FEW THAT PRESENT WITH EITHER BI-POLAR OR SCHIZOPHRENIA. THESE ARE NOW PSYCHIATRIC FEATURES. WHEN MOST IS INTACT JUST SLIGHTLY -- THE FUNCTION OF THE PROTEIN YOU'LL SEE MORE PSYCHIATRIC -- AND THIS IS THE FIRST THING I WANT TO BRING TO YOUR ATTENTION BECAUSE AS WE THINK ABOUT PSYCHIATRIC DISORDERS THEY ARE NOT GOING TO BE BAD OR SEVERE MU ADVERTISINGS. THEY WILL BE SUBTLE GENES. SEEING THE COMPLEXITY OF THE FENO TYPE -- WHAT ARE THE CONSEQUENCES OF LOSING THIS GENE IN DIFFERENT TYPES OF NEURONS. AND WE STUDIED MANY NEURONS AND I'M NOT GOING TO TAKE YOU THROUGH ANY OF THAT BUT I WANT TO TELL YOU ABOUT SOME SURPRISES THAT WE LEARNED AND BRING BACK THE IDEA OF WHAT MIGHT BE EXPECTED. WHEN WE DELETED THIS GENE -- THE SURPRISE TO US WAS WE GOT SOME OVERLAPPING FENO TYPES FOR BOTH WE SEE THE PARTIAL REDUCTION -- OR SIGNALING OR -- [ INAUDIBLE ] SIGNALING. BUT WHAT WE SEE IS WE NEVER SEE ANXIETY BUT WE SEE IT IN THE -- KNOCKOUT. WE NEVER SEE TREMOR. [ INAUDIBLE ] AND THIS WAS INTRIGUING AND WHEN WE SAW THAT WE DECIDED TO REALLY TEST BOTH LOSS AND RESCUE TO MAKE SURE THIS IS GOING TO HOLD AND I'LL SHOW YOU HERE SOME EXAMPLES SO WE USED -- WHERE YOU CAN DELETE THE GENE SO BASICALLY YOU ELIMINATE IT AND THEN THERE IS A SECOND WHICH IS PRACTICALLY -- BUT IF YOU AGAIN USE THE ENZYME -- NOT ONLY CAN BRING THE PROTEIN -- SO WHEN WE DO THIS YOU CAN SEE HERE -- MEASURINGtremor the -- which is a mild -- [ INAUDIBLE ] WHEREAS IF WE DO THE REVERSE -- NONE OF THAT HAPPENS. THE SAME IS ABOUT ANXIETY. IF WE MEASURE ANXIETY BUT HOW OFTEN THE ANIMALS SPEND TIME IN THE LIT SIDE OF THE DARK BOX. WHEREAS WHEN WE BRING THE GENE BACK WE RESCUE. THIS IS NOW THE NULL. WHY IS THIS? IT'S BECAUSE WHEN YOU HAVE THE DELETION OF THIS GENE -- AND WE'VE DONE A LOT OF PHYSIOLOGY -- WHAT WE LEARNED IS VERY PARTIALLY DISABLING THE NEURON. NOT TOTALLY DISABLING. THIS IS TELLING YOU WHAT WILL BE THE FIRST SYMPTOMS YOU MIGHT SEE FROM PARTIAL DYSFUNCTION. OF COURSE WITH AGE THE RESPIRATORY SYSTEM -- THEY WILL DIE. WE THEN ASKED AND WE NOW DELETE THIS GENE IN DIFFERENT SUBTYPES. WHAT MIGHT HAD HAD AND AGAIN WE SAW MODULARITY HERE. WE KNOW THAT MODERATE REDUCTION -- CAN CAUSE SEVERE NEURO PSYCHIATRIC -- AND WHAT I MEAN BY MODERATE I'M TALKING IN THE 30% RANGE REDUCTION. THIS LEADS ME TO BELIEVE THAT FOR ISOLATED TO SEE JUST AN ISOLATED ANXIETY IN A PERSON IN OCD WITHOUT THE OTHER SYMPTOMS IT'S GOING TO BE MUCH, MUCH MILDER. AND I THINK THIS IS SOMETHING THAT WE NEED TO KEEP IN MIND. MUTATION, WE USED TO THINK OF WE THINK OF AS BEING BENIGN. THOSE -- PARTIAL MUTATIONS THAT WE USED TO WRITE OFF AS UNIMPORTANT ACTUALLY MAY BE A RISK FACTOR AND THIS IS HOW WE THINK AS WE DEAL WITH NEW GENETIC DATA IN THESE DISORDERS. WE PRETTY MUCH TESTED ACTUALLY BOOSTING THE SYSTEM AND ALONE IT WAS NOT SUFFICIENT BECAUSE AS YOU CAN SEE THE ANIMALS WILL SUFFER. AND IT WAS THAT WORK TOGETHER AT THE SAME TIME WE'RE SEEING AN INCREASE IN THE NUMBER OF GENES AND DIVERSITY OF JEANS CAUSING INTELLECTUAL DISABILITY. THIS LED US TO EXPROPER A DIFFERENT APPROACH AS A PARALLEL TO THIS AND THAT IS WHEN WE THOUGHT MAYBE WE SHOULD BEGIN TO LOOK AT CIRCUIT STIMULATION SUCH AS BRAIN STIMULATION AND WE DECIDED TO FOCUS ON THE HIPPOCAMPUS BECAUSE WE KNEW THE ANIMALS AND THE PATIENTS WITH RHETT SYNDROME HAD PROBLMS WITH MEMORY PROBLEMS. HUMAN DATA STIMULATION OF THE DIAGONAL DEPARTMENT CAN ENHANCE ACTIVITY AND IN SOME CASES HAVE BEEN SHOWN TO INCREASE MEMORY. SO WE COLLABORATED -- AT OUR INSTITUTE WHO IS VERY ADAPT AT DOING THIS TECHNOLOGY AND RECORDED TO BE SURE THE ANIMALS ARE NOT EXPERIENCING ANY SEIZURES JUST SIMPLY STIMULATION AND THE PARADIGM WAS TO TAKE FEMALE RHETT SYNDROME MICE AND TO IMPLANT THEM WHEN THEY ARE ABOUT TWO MONTHS OLD AND PROVIDE DEEPER STIMULATION USING PARAMETERS IN THE NEURO SURGICAL SUITES FOR A COUPLE OF WEEKS AND THEN TEST FOR ALL BEHAVIORS AND PHENOTYPES THAT WE KNOW ARE IN THE HELP OH CAMPUS. I'M GOING TO SUMMARIZE AND SHOW YOU A COUPLE OF PIECES. SO THIS IS A CONTEXT WALLS SAY. THEY WILL GET A FOOT SHOCK. IF BROUGHT BACK THEY WHEN FREEZE BUT IF THEY HEAR THE SOUND IN A NEW CONTEXT THIS IS CUE DEPENDENT NOW. THESE ANIMALS DON'T HAVE A PHENOTYPE NOW. THEY HAVE A CONTEXT WALL FEAR. I'M GOING TO SHOW YOU WHEREBY THE DIRECT MICE NOW RECORDING THEIR LPT OVER MULTIPLE DAYS AND YOU'LL SEE THE DIRECT MICE HAS DECREASED AS SHOWN HERE. HOWEVER AFTER THE BRAIN STIMULATION THAT PRETTY MUCH NORMALIZED TO THE BASELINE. AGAIN I'M NOT SHOWING ALL OF THE CONTROLS BUT THEY IMPROVED SOME AS WELL. IN ADDITION WE FOUND THAT THE DIRECT MICE HAVE INCREASED NEUROGENESIS AND THAT REMARKABLY INCREASED AND INCLUDING THE NEURONS -- [ INDISCERNIBLE ] SO HAVING FOUND THIS WE WERE MORE WORK TO BE DONE. WE NEEDED TO KNOW HOW OFTEN THIS HAS TO BE DONE. HOW LONG IS THE EFFECT. I CAN ANSWER THIS QUESTION FOR YOU. WHEN WE DID THIS FOR TWO WEEKS AND TESTED THEM THREE WEEKS LATER THEY IMPROVED. AGAIN SIX WEEKS LATER THEY ARE STILL NORMAL BUT BEYOND SIX WEEKS THE EFFECT IS LOST. WE HAVE TO DO THIS EVERY SIX 2007 WEEKS. COULD THIS WORK ON OTHER MODELS. -- TESTED THIS ON OTHER MODELS AND BEGINNING TO SEE POSITIVE RESULTS. HOW IS TBS REALLY FIXING THE BEHAVIOR. THE PHENOTYPE. IMPROVING NEUROGENESIS. SO TO ANSWER THIS QUESTION A GRADUATE STUDENT IN OUR LAB AMY TOOK ON THE BECAUSE -- [ INDISCERNIBLE ] INVOLVED IN GENE AND SPECIAL REGULATION SHE WANTED TO SEE WHAT HAPPENS TO GENE EXPRESSION AFTER THE BRAIN STIMULATION. SO SHE APPLIED THE SAME PARADIGM THAT WAS APPLIED BUT SHE WANTED TO LOOK ACUTELY AT WHAT HAPPENED. AND I SHOULD MENTION TO YOU SOME THAT ARE ALTERED AND SOME ARE DOWN, MOST ARE DOWN AND THOSE ARE THE ONES THAT ARE CHANGED BY MORE THAN 20%. AFTER THE BRAIN STIMULATION WHAT WE FOUND IS THAT 25% OF THESE GENES ARE CORRECTED. WHICH IS QUITE A NICE NUMBER PARTICULARLY IF I TELL YOU THAT MANY OF THOSE GENES ARE IMPORTANT FOR SYNAPTIC FUNCTION AND NEUROGENESIS WHICH MIGHT EXPLAIN WHY WE SAW THE NEURO BEGIN SIS EFFECT CORRECTED. [ INDISCERNIBLE ] AFTER THE BRAIN STIMULATION I HOPE YOU CAN APPRECIATE THEY ARE ALL INTERMINGLED. SO THIS WAS QUITE INTERESTING AND BECAUSE -- HAS ALREADY SEEN SOME IMPROVEMENT IN ANOTHER MODEL OF INTELLECTUAL DISABILITIES WE WANTED TO SEE IF THERE ARE OTHER MODELS WHERE PEOPLE HAVE IN INTERPRET GATED THE SAME BRAIN REGION. AND ASKED -- WHICH GENES ARE ALTERED IN THESE MODELS AND ARE IN OF THE GENES CORRECTED OR BOOSTED. AND WHAT WE FOUND IS THAT IN TWO MODELS BECAUSE THERE ARE NOT MANY BUT THESE TWO CAUSE INTELLECTUAL DISABILITY ABOUT A QUARTER OF THE GENES THAT ARE DOWN REGULATED WE SAW THOSE ARE AMONG THE GENES THAT ARE TYPICALLY BOOSTED BY BRAIN STIMULATION AND THERE WAS DATA FROM PEOPLE WHO DIED FROM MAJOR DEPRESSION. AGAIN WE FOUND THAT MANY OF THESE HAVE BEEN CORRECTED AS WELL. SO WITH THAT WE THEN WANTED TO ASK WHAT HAPPENS WITH THE DIRECT MICE. SO FOR THAT WE USED THE SAME PARADIGM -- WE DID THE DBS AND THEN WE WAITED FOUR WEEKS AND WE KNEW ANIMALS HAD IMPROVED BEHAVIOR AND ASKED DID WE SEE -- WILL IT LAST THAT LONG? AND AGAIN HERE YOU'LL SEE THAT THE. [ INDISCERNIBLE ] ANIMALS HAVE MANY GENE -- [ INDISCERNIBLE ] SOME TRENDING AND SOME WITH WERE SIGNIFICANT. AT LEAST AT THE MOLECULAR LEVEL WE KNOW WE'RE BOOSTING MANY GENES THAT ARE IMPORTANT FOR DEVELOPMENT. CERTAINLY WE'VE SEEN IMPROVEMENT IN THE -- [ INDISCERNIBLE ] AND I SHOWED YOU THE MOLECULAR DATA SO THIS MIGHT HOLD A PROMISE. THIS IS MOLECULAR. HOW ABOUT THE NETWORK? I MENTIONED TO YOU THAT THE LTP WAS IMPROVED BUT EXACTLY HOW THAT IS WHERE WE BEGAN THE STUDIES TO LOOK AT THE BEHAVIOR OF CA1 NEURONS AND THIS IS STILL ONGOING WORK. SO I'M GOING TO SHARE WITH YOU SOME PUBLISHED WORK AND UNPUBLISHED WORK. WE FIRST STARTED USING CALCIUM -- INITIALLY IN SLICES FROM THE MUTANT MICE. AND I HOPE YOU CAN SEE THIS ON THE SCREEN. IF YOU LOOK IN WILD TYPE ANIMALS YOU'LL SEE SPARSE ACTIVITY AND YOU'LL SEE IT INDEPENDENT AMONG DIFFERENT NEURONS BUT IN THE MUTANT YOU'LL SEE HERE THERE ARE MORE NEURONS THAT FIRE TOGETHER AND I SHALL A SURE YOU WE'VE DONE THIS AT THE TIME THE ANIMALS DON'T HAVE SEIZURES. AND SHOWED THAT THIS INCREASE CO-FIRING OR INCREASED -- TOGETHER HAPPENS INDEPENDENT OF SEIZURE ACTIVITY. AND YOU'LL SEE THAT DATA HERE FOR SOME NEURONS. THE NULL -- MALES HAVE NEURONS FIRING TOGETHER BUT IN THE FEMALES THAT THE MOSAIC HAS MORE OF THE ENHANCED SYMPHONY. WE BEGIN TO -- TO TRY TO UNDERSTAND WHAT MAY BE DRIVING THIS AND WE'VE LOOKED AT MANY, MANY OF THE CONNECTIONS. I'M JUST SUMMARIZING HERE TO TELL YOU THE ONE DEFECT WE SAW WAS DECREASEED IMPORT -- YOU'LL SEE HERE ON THESE NEURONS -- [ INDISCERNIBLE ] WERE DECREASED AND WHEN WE THEN PERFORM DBS AND REPEATED THE STUDIES FIRST WE FOUND THAT THE ENHANCED SYMPHONY WAS NORMAL NORMALIZED AND WHEN WE REPEATED THESE STUDIES THIS TIME LABELING BOTH THE WILD TYPE NEURONS OR MUTANT NEURONS WHICH WE KNEE WERE RECORDING FROM BECAUSE THEY WERE MOSAIC. IN THE SHARED MICE THERE IS DECREASE IN THE FREQUENCY OF THE SPONTANEOUS EPSCs WHEREAS THE WILD TYPE CELLS WERE NORMAL AND THE AMPLITUDE ALSO CORRECTED. THIS IS IN SLICES AND THIS GAVE US A FOUNDATION AND WE'VE DONE RECORDING AND WE SAW A SIMILAR PATTERN OF FIRING AMONG THE NEURONS SO THAT LED US TO NOW GO TO -- TO RECORD FROM THESE ANIMALS AGAIN LOOKING AT THIS -- HERE USING THE MICRO ENDOSCOPE AND WHAT WE WANTED TO DO WAS DO THIS IN AWAKE BEHAVING ANIMALS BECAUSE NOW WE WANT TO -- [ INDISCERNIBLE ] AND SEE HOW DO THESE NEURONS BEHAVE. AND YOU'LL SEE HERE THAT FOR INDIVIDUAL CELLS AND THIS IS THE ESSAY WE DID. WE WENT BACK TO THE CONTEXTUAL FEAR WHICH THESE ANIMALS SOLIDLY HAVE A DEFICIT IN AND WE MONITORED THE NEURONS DURING TRAINING. THEN ONE HOUR LATER TO TEST THEIR SHORT-TERM MEMORY AND THEN ONE DAY LATER. WHEN WE KNOW THEY HAVE IMPAIRMENT AND THE FIRST THING WE REPRODUCED IN THESE DATA WAS THE ENHANCED SYMPHONY. THAT IS THE PHENOTYPE WE CONSISTENTLY SEE. NOW WE CAN LOOK A THE FIRING PATTERNS OF THE NEURONS AND ASK HOW DOES THAT CORRELATE TO THE ONE HOUR GOOD PERFORMANCE VERSUS ONE DAY NOT AS GOOD MEMORY CONSOLIDATION AND I JUST DO HERE FOR SIMPLICITY AND ONLY FOCUS ON THE CELLS THAT FIRED AND WILD TYPE ANIMALS DURING TRAINING AND TESTS. AFTER THE LEARNING TESTS THAT IS WHEN YOU WANT THE SYNCHRONY TO HAPPEN. THE SAME NEURONS THAT FIRE WHERE THE TRAINING WAS OBTAINED THERE HAS TO BE AN ENSEMBLE THE NEXT DAY WHEN THEY ARE PUT IN THE TEST. BUT IN THE RHETT ANIMALS WE DID NOT SEE THAT. WE SAW MUCH FEWER OVERLAP. ESPECIALLY WHEN THEY HAVE THE PHENOTYPES. AT ONE HOUR YOU DO SEE AN OVERLAP BUT BY ONE DAY THAT IS QUITE DIFFERENT. SO NOW THIS GAVE US TOGETHER THE DATA FROM THE IMAGING DURING THE BEHAVIOR TOGETHER WITH OUR SYNAPTIC STUDIES GAVE US GROUNDS ON WHICH NOW WE'RE BUILDING. WE WANT TO NOW REALLY KNOW IF THE SYNCHRONY IS INTERFERING WITH THE LEARNING AND THE WAY THAT WE'RE GOING TO DO THAT IS TO SEE IF WE CAN INDUCE IT IN HEALTH IAN MALLS AND SEE IF THAT WILL LEAD TO HEALTHY LEARNING. -- WE WANT TO SEE IF THE -- [ INDISCERNIBLE ] ON THESE ORGANS. [ INDISCERNIBLE ] NEURONS IS ONE OF THE WAYS BY WHICH DBS IS WORKING SO WE'RE GOING TO TRY TO STIMULATE THESE -- CONNECTIONS AND SEE IF THAT ALSO WILL CORRECT THE PHENOTYPE AND THESE ARE SOME EXAMPLES AND WE REALLY BENEFIT FROM THE TOOLS AND WE'RE GRATEFUL FOR THAT. MOVING FROM RETT I WANT TO MOVE TO THE OTHER SIDE OF THE COIN THE OTHER DISORDERS THAT IS ALSO CAUSED BY MUTATIONS IN THE SAME GENE. THE WAY I DISCOVERED IT HAPPENED FIRST IN THE MOUSE. MY FIRST EXPERIENCE AFTER FINDING THE GENE AND FINDING THE LOSS OF FUNCTION -- WAS TO PUT THE GENE BACK AND SEE IF WE COULD RESCUE. WE TOOK THE GENE AND. [ INDISCERNIBLE ] BECAUSE THAT PIECE OF DNA HAD ALL OF THE NECESSARY AND HAD NO OTHER GENES NOTHING ELSE. SIMPLY HAD THE GENE THE SPLICING AND EVERYTHING. WE PUT THAT HUMAN GENES IN THE MOUSE AND THEN WE WERE GOING TO BREED IT TO THE -- MOUSE BUT TO OUR SURPRISE THE MOUSE HAD MULTIPLE PHENOTYPES. LEARNING AND MEMORY PROBLEMS AND EARLY DEATH AND THAT MOUSE HAD TWICE THE NORMAL LEVEL OF PRE TEEN. AND WE MEASURED THAT MANY HAVE LOOKED FOR THEM AND THIS IS HOW THE SYNDROME BECAME POPULAR AND RECOGNIZED BECAUSE PEOPLE REALIZED DOUBLING THIS GENE IN A HUMAN CAN CAUSE A DISEASE AND EVERY FEATURE OF THE HUMAN THEY'VE PREDICTED AND SO DO SOME OF THE HUMANS. WE DID GENETIC EXPERIMENT WHERE WE KNEW THAT THESE ANIMALS HAD THE FEATURES AND WE ASKED IF WE NORMALIZE GENETICALLY WE GOT RID OF ONE OF THE GENES -- COULD WE RESCUE THE SYMPTOMS AND WE RESCUED ALL OF THE SYMPTOMS IN ADULT MICE. WE WANTED TO USE A MOLECULE THAT WE COULD TAKE TO THE HUMANS AND THIS IS WHY WE COLLABORATE. [ INDISCERNIBLE ] AND I'M SURE YOU'VE HEARD ABOUT THEIR WORK USING THAT TO TREAT SPINAL MUSCULAR ATROPHY. THERE IS ONE COMPANY THAT HAS GONE IN INFANT AND WE SHOULD THINK ABOUT GOING TO INFANTS TO TREAT AS EARLY AS POSSIBLE FOR BETTER RECOVERY. SO WE COLLABORATED WITH THEM AND THEY MADE THE HUMAN GENE SO WHEN WE PUT THIS IN THE VENTRICLE OF THE MICE IT GOES EVERYWHERE AND NORMALIZES THE LEVEL OF THE PROTEIN. AND THE SINGLE CELL LEVEL YOU NORMALIZE THE LEVEL OF THE PROTEIN AND I'M NOT SHOWING YOU ALL OF THE BEHAVIOR BUT ALL. BEHAVIOR IS CORECTED. THESE ANIMALS ARE VERY ANXIOUS. THE MOST BEAUTIFUL RESULT I AM AS NEUROLOGIST -- THEY HAVE CONSTANT SEIZURES UNTIL THEY ARE SEVEN MONTHS OLD. THEIR EEGs NORMALIZE AS YOU CAN SEE HERE AND ALL. SEIZURES STOPPED. SO THIS IS EXCITING WHEN YOU START A TREATMENT IN A SEVEN-MONTH-OLD MOUSE WHO IS SYSTEMATIC FROM FIVE WEEKS OF AGE. AT LEAST IT'S HOPEFULLY. SO WITH THAT WE DECIDED NOW IT'S TIME TO TAKE THIS REALLY FOR A HUMAN TRIAL AND OF COURSE THERE IS A PROBLEM. BECAUSE I TOLD YOU HOW IMPORTANT MY P2 LEVEL ARE. SO THEN WE HAVE TO SEE IF THE ASO CAN BE TIED TO IT. WE CREATED A NEW MOUSE WHERE WE USED OUR FIRST HUMAN KEEP BUT THEN WE CREATED A SECOND ONE. [ INDISCERNIBLE ] AND SHOWED THAT THESE MICE HAVE TWICE THE NORMAL OF MP2 AND HAVE ALL. BEHAVIOR OF THE ORIGINAL MICE AND THEN WE BEGAN TESTING. YOU CAN SEE IF YOU USE A CONTROL ASO IT'S NOT TARGETING -- BUT IF YOU. [ INDISCERNIBLE ] DEPENDING ON THE DOSE AND THE DOSE DEPENDENT MANNER REDUCE THE LEVEL OF THE PROTEIN. THIS IS TOO LOW. THIS IS SIMILAR SO THIS IS A GOOD ONE SO WE USE THIS DOSE THEN TO GO BACK AND TREAT THE ANIMALS AND HERE IS AN EXAMPLE OF ONE PHENOTYPE. THESE ANIMALS HAVE EXCESSIVE FREEZING IN THE CONTEXTUAL FEAR BUT WITH A HIGH DOSE OF NORMAL. SO WE'RE NOW FINISHING THE STUDIES AT LEAST NOW WE KNOW WE CAN -- [ INDISCERNIBLE ] AND WE HOPE TO START WITH THE HUMAN WITH THE LOWEST POSSIBLE AND LOOKING FOR A PERIPHERAL BIOMARKER LOOKING IN THE BLOOD TO FIND SOMETHING THAT WILL REFLECT WHAT IS HAPPENING AND I THINK WE HAVE ONE BUT I DON'T WANT TO SHOW IT. IT'S TOO EARLY BUT IT WILL BE GREAT IF WE CAN MONITOR -- BASED ON THE BLOOD TESTS. SO THIS IS WHERE WE ARE AND I THINK THIS IS WHERE THE ANIMAL MODEL HAS BEEN HELPFUL. ONE TO DISCOVER THE DISEASE AND SECOND FOR US TO TRY A PARTICULAR THERAPY. THIS IS JUST A. [ INDISCERNIBLE ] I JUST WANT TO SUMMARIZE WHAT I LEARNED FROM STUDYING. THAT LEVELS REALLY MATTER. HERE IS THE -- DYE HERE IS THE RETT MOSAIC. [ INDISCERNIBLE ] EITHER SCHIZOPHRENIA OR BI-POLAR. THE SAME MU ADVERTISING CAN CAUSE BOTH WHICH IS REALLY INTERESTING. THIS IS THE FIRST PROTEIN THAT MADE ME RESPECT PROTEIN LEVELS. AND IT WAS THE STUDY ON DUPLICATION THAT INSPIRED US TO LOOK AT ANOTHER PROTEIN. SHANK 3 WHICH MANY OF YOU ARE FAMILIAR W MANY OF YOU HERE IN THE ROOM HAVE STUDIED THIS PROTEIN. CONNECTING MANY SYNAPTIC -- [ INDISCERNIBLE ] AND IT'S BEEN KNOWN STUDIED BY MANY CLINICIANS AND SCIENTISTS -- THE SYNDROME. THAT IS A SEVERE -- WHERE LOSS OF ONE ACCOUNTS FOR 1% OF AUTISM AND INTELLECTUAL DISABILITY. IT'S A SEVERE SYNDROME AND SOMETIMES PSYCHIATRIC DISORDERS. WHAT I WAS INTERESTED IN IS THE DUPLICATION. THEY WERE BIG SPANNING MULTIPLE GENES BUT THEY PRESENTED WITH ADHD -- RESTLESSNESS. BAD TEMPERA SCHIZOPHRENIA AND THE REASON I WAS INTERESTED. THERE WERE TWO CASES OR THREE REPORTED IS BECAUSE OF THE ADHD. AS NEUROLOGIST THIS IS A COMMON PROBLEM IN OUR SPECIALTY AND COULD IT BE IF ALL OF THESE GENES SHANK 3 IS CAUSING ADHD AND CAN WE PROVE THAT. WE TAGGED IT AND CREATED A MOUSE THAT HAD JUST ONE EXTRA SO THE MOUSE WILL HAVE 50% MORE. IT'S THE ONE THAT JUST SIMPLY CAN'T SIT STILL AND THAT IS THE PHENOTYPE OF THE MOUSE. THIS MOUSE WILL ALWAYS BE ON THE GO AND IF WE QUANTIFY THEIR ACTIVITY BOTH MALES AND FEMALES WITH DISTANCE TRAVEL AND THE ACTIVITY IS CLEARLY INCREASED SO WE THOUGHT THIS WAS ACTUALLY ADHD AND GOT VERY EXCITED BUT WHEN WE GAVE THEM AM FENCE MEAN THEY GOT EXTRA HYPER. I HAVE A SON WHO WAS GOING INTO PSYCHIATRY. NO WAY IN EARTH CAN YOU USE THE TERM MANIA WITH MICE. THE ONLY ONE EXPRESSED IN BRAIN THAT IS DUPLICATED IS SHANK 3 AND THESE PEOPLE HAVE BI-POLAR TYPE. ONE OF THEM WAS DIAGNOSED WITH ADHD BUT WAS ALSO RESISTANT TO AMPHETAMINES. BUT IN THE HUMANS WE LEARNED THERE WERE SEIZURES AND WE HAVE NOT PAID ATTENTION FOR SEIZURES SO THAT LED US TO GO BARK TO THE MICE AND THESE MILES ALSO HAD SEIZURES AND I WILL NOT GET INTO THE MECHANISM HERE. WE SHOWED THAT THESE ANIMALS HAVE INCREASED -- [ INDISCERNIBLE ] SYNAPSIS. WHEN WE TESTED LITHIUM ON THEM THEY DID NOT RESPOND BUT WHEN WE TREATED THEM WITH. [ INDISCERNIBLE ] ALL. FEATURES NORMALIZED INCLUDING THE EEGs. SO STARTING WITH THE MOUSE WE FOUND A PHENOTYPE, MADE US GO BACK AND LOOK AT HUMAN. THEY TOLD US ABOUT SOME OTHER FEATURES AND THEN WE TESTED THE TREATMENT THAT AT LEAST FOR THESE PATIENTS NOW THIS IS THE BETTER TREATMENT BECAUSE THEY PATIENTS DON'T TYPICALLY RESPOND TO SOME OF THE OTHER DRUGS. WE CONTINUED TO DO A LITTLE BIT OF BIOCHEMICAL WORK WITH SHANK 3 AND I BECAME INTERESTED IN ONE RARE MUTATION. THIS IS SENSITIVE TO LEVELS. WHERE YOU HAVE TO HAVE THE TWO. WE KNOW THE DUPLICATION -- BUT THERE WAS ONE MUTATION WE BECAME INTERESTED IN. IT WAS ONLY SEEN IN ONE PATIENT. BUT WE -- IT'S HARD TO MAKE A CONCLUSION AFTER ONE PATIENT. WE WERE INTERESTED BECAUSE WE LOOKED AT THE -- [ INDISCERNIBLE ] OF SHANK 3 AND WE FOUND -- OF COURSE CORRELATED AND IT IS THIS -- THAT IS MUTATED -- [ INDISCERNIBLE ] AND IT IS IN A DOMAIN THAT IS IMPORTANT FOR INTERACTION WITH ABLE INTERACTING ONE. THIS IS A PROTEIN THAT -- TO SHANK 3. SO WE DECIDED THIS IS WORTH IT. WE CAN'T BE SURE IF THIS IS CAUSING THE PHENOTYPE. THE ONLY WAY WE WILL KNOW IS IF WE STUDY THIS MORE. WE MADE THE EXACT SAME MUTATION AS IN THE MOUSE. AND WHEN WE MADE THAT AND WE -- PRECIPITATED IBA1 WE FOUND THAT THE SHANK 3 INTERACTS LESS. SO WE CONFIRMED THIS IS IMPORTANT FOR THE INTERACTION AND YOU SEE THE QUANTIFICATION HERE BUT WHEN WE LOOKED AT OTHER PARTNERS OF SHANK 3 THEY INTERACT NORMALLY AND WE TESTED MANY OF THEM. THIS WAS REALLY NEAT. AND NOW THE QUESTION DOES THAT CAUSE ANY PROBLEM. AND OTHERS HAVE SHOWN IN THE SHANK 3 -- HOWEVER WE ONLY FOUND THREE PHENOTYPES THAT ARE MORE RELEVANT TO SOCIAL BEHAVIOR. ONE IS AN ABNORMAL GROOMING PATTERN. THESE MICE YOU'LL SEE THAT THEY ARE FINE TOGETHER. THEY MIGHT GROOM EACH OTHER A LITTLE BIT BUT NOT MUCH WHEREAS THIS MOUSE YOU'LL NOTICE IS NOT GOING TO LEAVE THE INTRUDER. IT'S GOING TO BE CONSTANTLY GROOMING THIS INTRUDER AND YOU CAN SIT AND WATCH AS LONG AS YOU WANT. THAT IS WHAT THEY DO AND IF YOU ARE TO LOOK AT THE QUANTIFICATION. THE SECOND PHENOTYPE, THEY HAVE TYPICALLY A MOUSE WILL BACK OUT. HALF OF THE TIME ONE OF THE MICE WILL BACK OUT. THESE DON'T BACK OUT. EITHER THEY DON'T RECOGNITION OR THEY ARE STUBBORN OR DOMINANT. WE DON'T KNOW HOW TO DETERMINE IT. SO THESE WERE THE ONLY -- WE STUDIED MANY BEHAVIORS. THESE WERE THE ONLY BEHAVIORS AND THEY SOMEHOW HAVE TO DO WITH PERHAPS SOME SOCIAL BEHAVIOR IN MICE AND THESE MICE ALSO HAVE DECREASED SYNAPTIC TRANSMISSION AND -- SPINE DENSITY. HERE WE LOOKED BECAUSE SHANK 3 IS THE PREDOMINANT SHANK AND YOU'LL SEE BOTH THE FREQUENCY AND THE AMPLITUDE OF THESE -- REDUCED AND THEY HAVE ALSO SOME REDUCTION IN THE SPINE DENSITY. SO BECAUSE I MENTIONED TO YOU AB I1 IS IMPORTANT FOR BRINGING LINK CAN TO SHANK 3 WE DECIDED TO PUSH FURTHER FOR THE BIOCHEMISTRY AND WE LOOKED AT THE INTERACTING PROTEINS FOR AB I1 AND WE'VE IDENTIFIED THE INTERACTING PROTEIN OF SHANK 3 AND IN RETT YOU SEE SOME OF THOSE SHARED IN COMMON INCLUDING THE WAVE COMPLEX THAT IS CRITICAL FOR -- [ INDISCERNIBLE ] AT THE SPINE FOR THE SPINE FORMATION AND WHAT WE FOUND IN THESE MUTANTS -- WE DO NOT BRING THE WAVE COMPLEX WHICH IS CRITICAL. AND THIS SUGGESTED TO US THAT THIS IS ABI -- BRINGING THE WAVE ONE COMPLEX SO WE TESTED THAT AND WHAT I WANT TO SHARE WITH YOU HERE IS IF WE WERE TO TAKE BRAIN -- AND LOOK AT THE INTERACTION BETWEEN WAVE AND ABI -- AND IN SHANK 3 YOU'LL SEE THEY ARE PRETTY SIMILAR BUT THE ONLY TIME YOU SEE THE REDUCTION IF YOU LOOK -- [ INDISCERNIBLE ] SO I SHARE THIS STORY WITH YOU AND GO THROUGH ALL OF THE BIOCHEMICAL DATA TO SHOW YOU HOW SUBTLE THE CHANGE CAN BE. IT CAN BE VERY SPECIFIC. PROTEIN SPECIFIC. REGION SPECIFIC. SORT OF THE SAME THEME THAT TOM PRESENTED US TO THIS MORNING. BUT IN A DIFFERENT PROTEIN IT'S LIKELY A DIFFERENT CHEMICAL REACTIONS SO IF WE WERE TO LOOK AT SHANK 3 IT'S A SIMILAR STORY -- WHERE. [ INDISCERNIBLE ] THE SYNDROME. THE VERY SEVERE -- [ INDISCERNIBLE ] GIST YOU ISOLATED AUTISM. AND OF COURSE -- I PREDICT THERE MAY BE MILD MUTATION RIGHT HERE IN BETWEEN AND THEN THE DUPLICATION OF COURSE CAUSES BI-POLAR. SO WHAT WE'VE LEARNED FROM THESE STUDIES YOU CAN EITHER ALTER THE LEVEL OF THE PROTEIN OR ACTIVITY SLIGHTLY AND EFFECT A FUNCTION AND GOES THROUGH A PSYCHIATRIC PHENOTYPES. I DIDN'T SHOW ALL OF THE DATA BUT THEY HAVE ENHANCED OR INCREASED SYNAPSIS. THIS IS THE NORMAL. AND OF COURSE THE 50% IS THE MOST SEVERE. SOMEHOW SOMEWHERE BETWEEN THE NORMAL AND WHAT WE'VE SEEN -- IS GOING TO BE WHAT MAY BE GIVING -- THE ADULT PSYCHIATRIC DISORDER. THE CHALLENGE IS -- [ INDISCERNIBLE ] HOW WE'RE GOING TO PROVE THEIR FUNCTIONAL SEQUENCES. WE WERE LUCKY THAT THE MICE GAVE US THAT. IF WE DO RIGOROUS EVALUATION OF THE MICE WE WILL SOMETIMES UNCOVER SOME OF THE PHENOTYPES AND I HOPE I CONVINCE YOU TODAY THAT GOING BACK BETWEEN HUMANS AND MICE WE'RE ABLE TO LEARN SO MUCH ABOUT THE IMPORTANCE OF THESE PROTEINS FOR NEURAL FUNCTION AND BE BEHAVIOR AND I HOPE WE WILL BE ABLE TO SOLVE SOME OF THESE DEVASTATE IS DISEASES IN THE YEARS TO COME INCLUDING THERAPY. IN CLOSES I WOULD LIKE TO THANK ALLFUL PEOPLE IN THE LAB THAT HAVE CONTRIBUTED. AND ARE SOME ACTIVELY STILL WORKING. AND OF COURSE WE HAVE OUR COLLABORATORS FOR THE D BRAIN STIMULATION AND SYNCHRONY AND COMPUTATIONAL DATE AND I'M VERY GRATEFUL AND THE RETT SYNDROME FOUNDATION AND OF COURSE HOWARD HUGHES. THANK YOU. [ APPLAUSE ] >> I WONDERED IF I COULD ASK YOU ABOUT THE SYNAPTIC PHENOTYPE AND NORMALIZATION. YOU TOLD US SOME STORIES ABOUT DEEP BRAIN STIMULATION. AND YOU ELUDED TO A PHENOTYPE IN TERMS OF SOME ALTERATION. COULD YOU LINK THOSE THINGS TOGETHER? DO YOU THINK THAT THERE IS A FUNDAMENTAL SYNAPTIC PHENOTYPE THAT IS RESTORATIVE OR DO YOU THINK YOU CAN COMPENSATE FOR THAT WITH ADDITIONAL NEURONS. >> SO YOUR QUESTION IS ARE THERE ADDITIONAL NEURONS WHAT IS HELPING OR IS THAT WHAT GAVE US NOW INCREASED SPONTANEOUS EPSCs ON THE INHIBITORY NEURONS OR IS IT REALLY THE ACTUAL CHANGE IN THE SYNAPSIS? SO, THAT IS A GOOD QUESTION. THE FIRST THING TO ANSWER THIS QUESTION WE ASKED OF THE NEW NEURONS THAT ARE BORN, OF COURSE A VERY SMALL PERCENTAGE OF THESE MAKE IT TO NEURONS. ARE THEY WILD TYPE OR MUTANT. SO WE'RE MAKING JUST AS MANY RETT NEURONS. AT THIS TIME I CANNOT SAY WHETHER THE LPT IMPROVEMENT AND THE RESTORATION OF NORMAL -- INHIBITORS IS DUE TO HAVING 5% MORE NEURONS VERSUS ACTUALLY THE NEURONS ARE MORE ROBUSTLY ACTIVATED. I BELIEVE IT IS THE LATTER BECAUSE THE CORRECTION IS BACK TO -- [ INDISCERNIBLE ] AND HAVE OF THEM ARE MUTANT. >> THAT IS A GREAT TALK. I HAVE A COUPLE OF QUESTIONS. DID YOU OBSERVE WITH YOUR SOCIAL -- THAT THE MILES WERE ALSO HYPER AGGRESSIVE TOWARDS OTHER MICE AND DID YOU LOOK AT WHETHER THEY HAD HIGH TESTOSTERONE. WHAT DO YOU THINK IS BEHIND THE HIGHER AGGRESSION, THE DOMINANCE PHENOTYPE. >> THAT'S AN INTERESTING QUESTION. WHAT DRIVES THE HYPER -- WHAT DRIVES THE NOT BACKING OUT. THAT IS A PHENOTYPE I SAW IN THE RETT MOUSE. SO IT'S NOT NORMALLY DUE TO THE -- MOUSE. PROGRESSION. WE DON'T SEE THAT. BUT I DON'T KNOW -- THAT THE TESTOSTERONE IS NORMAL. WE HAVE NOT MEASURED IT. THAT IS A GOOD IDEA. ACTUALLY THEY ARE DOCILE. THEY ARE NOT AGGRESSIVE. THEIR LACK OF BACKING OUT THEY JUST DON'T KNOW ANY BETTER. WE SEE THAT OF TERRET MICE. >> IS THERE A DIFFERENTIAL -- [ INDISCERNIBLE ] IN DIFFERENT PARTS OF THE BRAIN. >> DIFFERENT PARTS OF THE BRAIN? THAT IS A GOOD QUESTION. [ INDISCERNIBLE ] I HAVE NOT LOOKED CAREFULLY IN THE BRAIN REGION SPECIFIC MANNER. [ INDISCERNIBLE ] -- [ INDISCERNIBLE ] LET ME BACK PARAPHRASE THE QUESTION. WHEN WE TAKE IT OUT FROM -- [ INDISCERNIBLE ] MOTOR FROM -- DOPAMINE AND SO ON. SHE IS ASKING IN HUMAN GROWTH SINCE IT'S A MOSAIC DID YOU SOMETIMES SEE AN ISOLATED PHENOTYPE IF THE MOW SAYISM GO ONE WAY OR THE OTHER. WE SEE MANY GERMS WITH FAVORABLE X CHROMOSOME ACTIVATION WHICH MEAN HEALTHY SURVIVE -- AND THE 90% OF THE FUME RONES HAVING HEALTHY AND ONLY 10% HAVING THE MUTANT. [ INDISCERNIBLE ] THEY MAY PRESENT ONLY WITH AUTISM. TREMOR AND SLIGHT LEARNING ACTUALLY SOME OF THEM ARE MOTHERS. THEY GO ON AND GROW UP TO HAVE KIDS SO WE ASSUME PARTIAL -- -- I MENTIONED -- OBESITY SO THESE FEMALES THAT HAVE MUCH MILDER -- THEY EAT MORE. THEY ARE MORE OBESE. VERY DIFFERENT FROM THE CLASSIC RAT. >> DID YOU DO A DEEP BRAIN STIMULATION IN HUMANS? >> I HAVE NOT TRIED TO DO DEEP BRAIN STIMULATION IN HUMANS YET BUT I'M BEGINNING TO TALK TO OUR NEURO SURGEONS ABOUT IT. WHAT WE WANTED TO DO OUR WHOLE TEAM IS WE WANT TO SEE CHRONICALLY. IF YOU DO THIS FOR A YEAR DO YOU STILL GET THE SAME BENEFIT. THIS IS REALLY IMPORTANT TO DO A LITTLE BIT MORE PRECLINICAL WORK AND SOMETHNG I FORGOT TO MENTION BECAUSE WE FIND THAT THE TERRET BRAIN IS RESPONSIVE TO THIS KIND OF DEEP BRAIN STIMULATION WE'RE TESTING OTHER BRAIN AREAS BECAUSE THE MULTIPLE DYSFUNCTION IS DEBILITATING AND WE WANT TO SEE, WE PICKED THE HARDEST ONE TO ASSESS IN A CLINICAL TRIAL. IF YOU TAKE A 10-YEAR-OLD TERRET CHILD IT'S HARD TO SEE HOW MUCH THEY CAN LEARN QUICKLY BECAUSE THEY CANNOT TALK. BUT MOTOR YOU MIGHT BE ABLE TO. SO WE'RE NOW TESTING ANOTHER AREA OF THE BRAIN AND WE'RE BEGINNING TO SEE A POSITIVE RESULT. WE MAY DO MORE FIRST. [ APPLAUSE ]