IT'S ALWAYS AN HONOR WHEN YOU GET TO INTRODUCE SOMEONE THAT YOU TRAINED UNDER HAVE THE OPPORTUNITY TO INVITE SOMEONE YOU TRAINED UNDER, ESPECIALLY IF YOU GET ALONG WITH THAT PERSON. BUT FOR ME IT GOES BEYOND THAT BECAUSE SCOTT IS HERE TODAY, REALLY A HUNDRED PERCENT RESPONSIBLE FOR THE WAY THAT I THINK ABOUT MY SCIENCE. I KNOW THIS IS NOT SUPPOSED TO BE ABOUT ME BUT IT WILL BE. SO I STARTED ROTATING HIS LAB AND REALLY A COUPLE REASONS, BRAVE HEART WAS MY FAVORITE MOVIE IN HIGH SCHOOL AND THAT JOKE WILL MAKE SENSE WHEN HE STARTS SPEAKING AND SECONDLY HE WAS DOING AMAZING CUTTING EDGE NEUROSCIENCE RESEARCH, 1 OF THE FIRST PEOPLE WHO TAKE ADVANTAGE OF USING TOOLS TO MANIPULATE NEURAL C-TYPE LECTINS ANDIVITY WHICH WE ALL TAKE FOR GRANTED TODAY AND THEN 1 OF THE THINGS THAT REALLY DREW ME TO HIS LAB WAS HOW ENGAGING AND ENCOURAGING AND APPROACHABLE HE WAS TO HIS TRAININGS. AND I REMEMBER WHEN I STARTED IN HIS LAB, I THOUGHT I WOULD WALTZ IN AND GET MY Ph.D. BY GETTING THE COOLEST PERSON IN GRAD SCHOOL, BUT HE QUICKLY LET ME KNOW, HE WAS IN AGREALT THEY WAS THE COOLEST PERSON IN GRAD SCHOOL BUT HE TAUGHT ME THE MOST IMPORTANT THING TO BECOME AN ACCOMPLISHED SCIENTIST AND THAT WAS COMMITMENT. SO TO GIVE YOU A BRIEF HISTORY ABOUT SCOTT BECAUSE I WILL INTRODUCE HIM. HE DID HIS Ph.D. AT UNIVERSITY OF LONDON BEFORE SKIPPING CROSS THE POND, RIGHT? YEAH? TO M. I.T. WHERE HE STARTED OLFACTORY MEMORY AND HE REALLY PUBLISHED HIS SEMINOLE PAPER IN CELL IN 2001 WHICH HE USED TO LAUNCH HIS CAREER BECAUSE LATER THAT YEAR, HE STARTED HIS OWN GROUP AT UNIVERSITY OF MASSACHUSETTS MEDICAL SCHOOL WHERE HE STAYED WITH A DECADE AND 2011 HE MIXED UP THE POSITION AS A WELCOME TRUST, SENIOR RESEARCH FELLOW ATOX FORD WHERE HE'S BEEN EVER SINCE. OF COURSE I COULD STAND UP AND TELL YOU A MILLION STORIES OF WHAT IT WAS LIKE WORKING WITH SCOTT. I'LL MENTION A FEW, 1 WAS THAT HE COERCED ME ONCE TO WEAR THE BORAT MAN KKEY IN THE LAB WHICH WAS QUITE AN EXPERIENCE, SECOND THING THAT HE'S DONE, HE INTRODUCED ME TO SCOTCH WHICH RESULTED IN ME WAKING UP COVERED HEAD TO TOE IN DRIED BLOOD WILL TELL YOU MORE ABOUT THAT STORY SOMETIME BUT REALLY THE MOST IMPORTANTLY, REALLY HE'S BEEN AN INSPIRATION FOR MYSELF AND FOR A MEMBER OF HIS TRAINEES AND IT'S AN ABSOLUTE PLEASURE AND HONOR TO INTRODUCE HIM SO SCOTT, >> WELL, THANKS MIKE, I TAKE NO RESPONSIBILITY FOR ANY OF THESE STORIES. MY RECOLLECTION IS THAT MIKE TURNED UP AND ASKED TO BE A STUDENT IN MY LAB AND I TOLD HIM HE [INDISCERNIBLE]. OTHERWISE, YOU KNOW HE'S GOING TO FEATURE A BIT TOGETHER SO I'M NOT GOING TO [INDISCERNIBLE] ANYMORE. I THOUGHT I WOULD START I BIT OF ADVOCACY BECAUSE OF WHERE I AM, AS YOU CAN SEE FROM THE CLASSIC TIME HERE HE MARVELS AT WHAT THEY WILL ACHIEVE WAS A REALLY SMALL HAD CAPSULE AND HE KNEW THAT THOSE WHO HAVE SMALL BRAIN COULD DO AMAZING THINGS AND THE ABILITY TO DO PAPGHT REGRESSION, AND THEY'RE SOPHISTICATED INSECTS AND ABLE TO DO THINGS WITH A TINY BRAIN AND THIS SAME PRINCIPLE APPLIES TO EVERYTHING THAT MYSELF AND SEVERAL PEOPLE AT THE IN, IH THAT ARE IN THE AUDIENCE WORK ON WHICH IS DROSOPHILA AND THESE FLIES OF COURSE CAN DO INCREDIBLE THINGS, TOO WITH EVEN FEW NEURONS, 150,000 OR SO AND SO THE SAME PRINCIPLES THAT DARWIN REFERS TO RELATE TO WORK IN THE FLY. THE THINGS THEY CAN DO ARE ORCHESTRATED BY TINY NERVOUS SYSTEMS THAT ARE SOMETIMES CONTROLLED BY SINGLE PAIRS AND WHAT THIS PROVIDE SYSTEM AN OBVIOUS RESOLUTION TO THYSELF PROCESSES SO IF YOU CAN FIND THE RELEVANT NEURONS CAN YOU UNDERSTAND FAIRLY COMPLEX PROCESSES SUCH AS LAB AND MEMORY, THE LEVEL OF IDENTIFIABLE CELLS AND MY HOPE FOR A NUMBER OF YEARS WAS TO TRY TO IDENTIFY SOME OF THOSE CELLS SO A BIT OF WHAT WE WERE IN THE ROAD ABLE TO ADDRESS AND THE INTERVENTION TO THE RELEVANT PLACES WITH THE RELEVANT TIME AND I THINK WITH THE INCREDIBLE ADVANCES IN MICROSCOPY, IT'S NOT GOING TO BE TOO LONG BEFORE WE WATCH RELEVANT MOLECULES WITHIN THE RELEVANT CELLS OF THE BEHAVING ANIMAL. I THINK THAT TO ME WAS A DREAM WHEN I STARTED IN THE FIELD IN 96 OR SO, AND I THINK IT'S INCREDIBLE THAT ADVANCES HAVE BEEN MADE THAT THIS ACTUALLY NOW SEEMS ALMOST REALISTIC. THE TITLE OF THE TALK IS REALLY DO THE RIGHT THING. IT'S MY GENERAL LAB'S GOAL TO FRY AND UNDERSTAND HOW THE NERVOUS SYSTEM PROVIDES AN ANIMAL WITH THE CAPACITY TO DO THE RIGHT THING AT THE RIGHT TIME TO USE LEARNED INTREVERTIONZ AND USE IT IN THE SEMINOLE STATES TO DIRECT THE BEHAVIOR AT THE RIGHT TIME. SO FOR EXAMPLE, YOU KNOW WHERE YOUR FRIGE IS IN THE HOUSE AND YOU DON'T GO THERE TO FIND FOOD UNLESS YOU'RE HUNGRY OR UNLESS YOU HAVE SOME SORT OF EATING DISORDER, RIGHT? SO THERE ARE REASONS WHY YOU DO THINGS AT PARTICULAR TIMES AND OF COURSE WE KNOW THAT THIS COMES FROM INTERNAL STATE DEFICITS AND SO FORTH AND I'D LIKE TO UNDERSTAND THESE KINDS OF BEHAVIORIAL DECISIONS AT THE LEVEL OF IDENTIFIABLE CELLS. I ORIGINALLY GAVE THIS TITLE WHEN I WAS ASKED WHAT THE TITLE WOULD BE A COUPLE OF MONTHS AGO, I DECIDED I WON'T TALK ABOUT THIS. THIS IS PUBLISHED A COUPLE OF MONTHS AGO AND I DECIDED INSTEAD I WOULD TALK ABOUT SOMETHING BRAND NEW BECAUSE SOMETHING I'M REALLY EXCITED ABOUT THAT I HOPE YOU WILL ALSO FIND THAT INTERESTING, THAT IS MECHANISMS WITH MEMORY EVALUATION, THAT WILL BECOME EVIDENT OF WHAT THAT ACTUALLY MEANS AS I GO FORWARD. I ALTS LIKE TO ALMOST START BY SAYING THAT OF COURSE I DO VERY LITTLE OF THE REAL WORK NOWADAYS, I'M A CELLS MONITOR AND SPEAKER AND WRITER AND THE PEOPLE THAT DID THE WORK THAT I'M GOING TO TALK ABOUT TODAY ARE HIGHLIGHTED IN THIS SLIDE. MOST OF THE WORK I'LL TELL YOU ABOUT TODAY WAS PERFORMED BY JOHANNES, WITH ANOTHER WORK,ER P A OLA, AND MUCH OF THE WORK I WILL REFER TO IN THE BROG ECTOMYOSIN WAS PERFORMED BY OLIVER AND THE [INDISCERNIBLE] WAS PERFORMED BY DAVID IN PERLYNN AND THEN EMANUEL IS NOW IN THE JOB MARKET, WITH ME FOR SEVERAL YEARS. SO MANY WHIMSICAL [INDISCERNIBLE] WAS IN MY GROUP, I TRIED TO INSPIRE HIM TO REVISIT OLD BEHAVIORIAL PATTERNS FIRST DEVELOPED BY BRUCE [INDISCERNIBLE] A FORMER ADVISOR IN THE LAB IN THE LATE 80S AND IT WAS A FAIRLY ASSOCIATED CONDITIONING PATTERN WHERE A HUNDRED OR SO FLIES WOULD BE GIVEN AN ORDER FOLLOWED BY ANOTHER ORDER AND THEY MAKE THIS ASSAY WORK PROPERLY AND THIS LED TO, I GUESS THE REST OF HIS CAREER AND WHY HE'S 1 OF YOUR COLLEAGUES, I'M SORRY BUT BUT HE WAS REASONABLY SUCCESSFUL. AFTER THIS CONDITIONING OF COURSE, THE FLIES ARE GIB A STRAIGHT CHOICE AND 1 OF INTERESTING THING SYSTEM THAT IN A VERY INTERNAL STATE DEPENDENT THE FLIES APPROACH THE ORDER WHEN THEY'RE HUNGRY. ONE THING I WANT TO MAKE CLEAR IS THAT THE TEST, THE SUGAR ISN'T PRESSENT, IT'S A DISPLAY FROM ANIMAL THAT THEY WILL FIND FOOD IF THEY APPROACH THAT REALLY VAUNT ORDER. --RELEVANT ORDER. TO GIVE YOU A SENSE OF WHAT ASSAY, THEY'RE PERFORMED IN THE CHAMBER AND THE ORDER IS DRAWN THROUGH ON THE HOUSE VACUUM, THE WALLS LINED WITH THE SUGAR REINFORCER AND THEN AFTER THEY'RE TRANSFERRED WHERE THEY RECEIVE THE CHOICE TO THE 2 ORDERS PRESENTED DURING CONDITIONING AND THEY PARTITION ACCORDING TO THEIR PREFERENCE AND A TYPICAL MEMORY ASSAY, 65 OR HUNDRED OR SO, WILL GO 35 WAY AND WE CALCULATE IN A MATHEMAT CALIPER FORMANCE AND THAT'S ABOUT A .3 HERE. I WANT TO MAKE THIS CLEAR BECAUSE THAT'S THE GENERAL RANGE OF THE SCORES YOU'LL BE LOOKING AT WITH 1 VERY STARTEDLING EXCEPTION AND THERE ARE A NUMBER OF CONTROLS, THAT WE WILL DO THAT I WON'T BORE YOU WITH FOR THE REST OF THE PRESENTATION. NOW WITH THIS ASSAY, THE FIRST THING WE SHOWED WAS THAT A SINGLE 2 MINUTE PAIRING PROVIDED MEMORY HERE, THAT YOU CAN SEE EXTENDS FOR UP TO A DAY AND A HALF. AND FOR THE LAST BIT WE'VE BEEN TRYING TO UNDERSTAND WHAT GIVES RISE TO THIS MEDIATION OF LONG-TERM MEMORY. SOMETHING ELSE THAT SHOWED UP WAS THAT THE MEMORIES CONSOLIDATED VERY QUICKLY. IF YOU TAKE THE FLIES, AND YOU TRAIN THEM AND THEN IMMEDIATELY AFTER TRAINING, YOU TREAT THEM ON AN ICE BATH AND ANESTHETIZE THEM AND LET THEM RECOVER FOR 24 HOURS AND MEASURE THE PERFORMANCE YOU'LL SEE THIS AS A CATATROPHIC EFFECT, BUT WITHIN 2 HOURS, THIS IS COMPLETELY INCONSEQUENTIAL, THE CLASSIC DEMONSTRATION WAS RAPID CONSOLIDATION OF MEMORY. AGAIN WE'VE BEEN INTERESTED IN TRYING TO WORK OUT WHERE THAT KIND OF PROPERTY COMES FROM AND 1 INTERESTING THING I WANT TO BEHABE LOOR ON THE RAPID CONSOLIDATION IS THE REINFORCING PROPERTY. THAT MEMORY CONSOLIDATION AND REEVALUATION FUNCTIONS SO I NEED TO GIVE YOU THE BASIC INTRODUCTION TO THE SLIDE IN EFFECTOUS SYSTEM AND IT'S AN OLFACTORY AND SUGAR REINFORCEMENT ASSAY SO THE FLY SOCIETIES THE SUGAR REINFORCEMENT BY THE THIS IS DONE HERE AND THIS IN THEIR SENSORY PROJECTION SYSTEM FROM THERE, AT LEAST IN PART OF OUR MODULE NEURONS THIS GETS TO THE MUSHROOM BODY, THIS BLUE STRUCTURE. THE OLFACTORY IS SHOWN HERE ON THE FLY HEAD AND THEY SEND THOSE IN INTOLE LINES, THIS IN EVERGZ IS REALLY SHOWN AGAIN, THE PLUSH ROOM BODY NOW THIS IS A GOOD POINT TO POINT OUT WHEN HE JOINED MY LAB, 1 THING I HAD HIM DO WAS TO SEE WHETHER HE COULD--WEATHER OLFACTION WAS MEDIATED BY THE I WAS TRYING TO REMOVE THIS FROM THE FLIES AND I CAME BACK AFTER A COUPLE OF HOURS, HE SAID THESE GUYS ARE TAKING FOREVER, THESE GUYS ARE COVERED IN THOSE ANTENNA, AND HE THOUGHT THE PRISTLES WERE ANTENNA AND SO SHE WAS SHAVING THE FLY AND NOT SURPRISINGLY AFTER 2 HOURS UNDER CO2, AFFIXIATION, IT WAS DEAD AS WELL AS BALD. REGARDLESS, HE BECAME SUCCESSFUL. [LAUGHTER] THE BLUE BULB OF THE MUSHROOM BODY IS NOT JUST A BLUE BLOB BUT FOR THE PURPOSES OF THIS TALK, IS SHOWING THE CELL BODIES WITH THE VESICLE PROJECTIONS HERE AND 1 THING WE KNOW FROM THE WORK OF MANY OTHER PEOPLE INCLUDING MARK [INDISCERNIBLE] HERE IS THE ORDER REPRESENTATION IS IN THE CANYON CELL LED AND RELATIVELY SPARSE POPULATIONS OF THE OVERALL ENSEMBLE, SO FOR EXAMPLE, AN INDIVIDUAL WOULD ACTIVATE 40 OR SO NEURONS OF THE OVERALL COLLECTION GIVING A HUGE ASSOCIATION MATRIX WITHIN THE 5000 OR SO NEURONS. ONE OTHER THING WE KNOW AND I'M SKIPPING OVER CELLS AND OTHERS, THESE USE DIFFERENT SUBSETS, ANATOMICALLY DISCREET NEURONS THAT PROVIDE THE REINFORCING EFFECTS OF ELECTRIC SHOCK AND SO FORTH AND OTHERS THAT REPRESENT REINFORCING EFFECTS OF GOOD THINGS AND SUGAR REWARD AND [INDISCERNIBLE] AND SO FORTH. AND THIS SIMPLE ENOUGH MAY OF COURSE SUGGEST THAT THE MEMORIES ARE FORMED IN DIFFERENT PLACES, SOMETHING THAT APPEARS TO BE TRUE. ONE OTHER THING THAT'S MATCHING AND I WON'T TELL YOU TOO MUCH ABOUT IS 1 OF THE THING THAT REWARD LEARNING DOES IS IT REDUCES THE SPECIFIC ORDER DRIVE, AND SO EXCUSE THE NETWORK AWAY FROM GENERATING AVOIDANCE AND SAVORS APPROACH AND I WILL SHOW YOU EVIDENCE OF THAT. SO OF COURSE THE PREVIOUS LINE DIAGRAM WILL GIVE YOU ANY IDEA OF THE COMPLEX NEUROANATOMY AND 1 OF THE THINGS THAT COMES ABOUT IN RECENT YEARS FROM THE FARM PROJECT IS ENORMOUS COLLECTION OF THESE DRIVERS, LARGELY PREDICATED ON THE DEVELOPMENT FROM THEN [INDISCERNIBLE] TO USE SPLIT GULF TO LABEL VERY SPECIFIC SUBSETS AND HERE, ACTUALLY IS A RECONSTRUCTION MOVIE FROM THE [INDISCERNIBLE] PROJECT BUT HE SHOWS YOU INDIVIDUAL DOPA MINERGIC MUSHROOMS FROM THE BODY. WE KNEW FROM WORK MANY YEARS AGO THAT THIS ORGANIZATION WAS EFDEPT WITHIN THE MUSHROOM BODY AND THE SIMPLE IDEA WAS THAT FORM USUALLY FOLLOWS FUNCTION SO PRESUMABLY SOME OF THESE NEURONS DID DIFFERENT THINGS AND THAT'S 1 OF THE THINGS WE PURSUED VERY HEAVILY STARTING IN MIKE'S PHASE IN THE LAB. ONE OTHER THING ABOUT THIS MODULAR ARRANGEMENT OF THE TERMINALS ON THE MUSHROOM BODY FROM THE NEURON SYSTEM THAT EACH 1 OF THESE ZONE SYSTEM ACTUALLY OCCUPIED BY CORRESPONDING SUBSET OF MUSHROOM BODY OUTPUT NEURONS SHOWN IN EQUIVALENT COLORS AND DENDRITIC CELL RIDDIC FIELDS OF THE DIFFERENT OUTPUT NEURONS, SUGGESTING THAT ACTUALLY THE DOPA MINERGIC NEURONS INNOIVATING EACH OF THESE ZONES ARE MODULATING THESE IN THE DIFFERENT OUTPUT PATHWAY AND LEADING TO THIS KIND OF MODEL CAME UP WITH SOMETIME AGO. SO IF EACH 1 OF THESE DIFFERENT CLASSES OF DOPA MINERGIC NEURONS AND THE OUTPUT NEURONS FROM THE CORRESPONDING ZONES KIND OF LOOP LIKE THIS. SO 1 THING WE KNOW IS THAT MANY OF THE REWARDING DOPA MINERGIC NEURONS INNOVATE THE HORIZONTAL TIPS AND THE NEURONS ARE LARGELYOT VERTICAL STOCK OF THE BODY AND IT TURNS OUT THAT ACTUALLY THAT OUTPUT NEURONS IN THESE DIFFERENT ZONES, THEY'RE ALSO DIFFERENT FLAVORS. THOSE THAT LEAVE THE HORIZONTA TIP DRIVE THE AVOIDANCE BEHAVIORS AND THOSE THAT DRIVE AND LEAVE THE VERTICAL LOBES DRIVE APPROACH BEHAVIORS AND AS I SAID ON THE PREVIOUS SLIDE, IT APPEARS THAT PUNISHMENT REDUCES THE AUTODRIVE TO THESE APPROACH PATHWAYS, THEREFORE PUTTING IT TOWARDS AVOIDANCE WHERE THE REWARDING DOPA MINERGIC NEURONS SILENCE THE OUTPUTS, THE AVOIDANCE MUSHROOM NEURONS IF THE APPROACH WILL SHOW IN THE GENERAL THING. SO FOR THE PURPOSE OF THIS TALK AND 1 THING THAT WILL COME UP AGAIN, THE KEY IS THAT AFTER REWARD LEARNING, THE DRIVE FROM ORDER TO THESE AVOIDANCE DIRECT MEASURE ROOM BODY IN THE NEURONS IS REDUCES. I'LL GIVE YOU A FEW EXAMPLES OF EXPERIMENTS THAT LED US TO THIS CONCLUSION FIRST. HERE IS A RECONSTRUCTION OF SOME OF THE REWARD IN DOPA MINERGIC SHOWN IN GREEN AND YOU CAN SEE THAT THE CELL BODIES LINE THE OUTSIDE AND THEY INNERIVATE THIS GRAY CLOUD WHICH IS THE MUSHROOM BODY ENSEMBLE AND MOST OF THE INNERIVATION IS IN THE VERY DENSE AREA ON THE TIPS OF THE HORIZONTA LOBES OF THE MUSHROOM BODY. INCIDENTALLY. THESE BLUE MIGHT OOH DENTIFY THESE AT THE STATE OF CONTROL OF THE REPET HIV MEMORY EXPRESSION AND YOU CAN SEE THAT THESE ARE VERY DISCREET REGIONS FROM THOSE THAT ARE WEB CONNECTED WARDED AND SO 1 OF THE THINGS THAT CAME UP IN THOSE DAYS WAS THE QUESTION OF HOW DOES THE STATE DEPENDENT SIGNAL THAT'S DELIVERED TO THE REGION ACTUALLY IMPACT MEMORY EXPRESSION FROM A LOCATION ELSEWHERE IN THE MUSHROOM BODY AND THE PAPER THAT I WAS GOING TO TELL YOU, BUT WON'T, WE FIND OUT THIS IS MEDIATED BY A SUBSET OF AND REGULATES THE AMOUNT OF FEET FORWARD IN THE NETWORK TO CONTROL WHETHER MEMORY IS EXPRESSED FROM THIS MORE DISTANT LOCATION. OF COURSE WE KNOW THAT THESE NEURONS PROVIDE REWARD IN TEACHING SIGNALS BECAUSE WE CAN DIRECTLY MANIPULATE THE FUNCTION AND ASK WHEN ARE THEY REQUIRED AND WHAT KIND OF PROPERTIES CAN THEY CONVEY IF YOU ARTIFICIALLY TRIGGER THEM. SO FOR EXAMPLE, IN SOME OF THE EXPERIMENTS WE ACTIVATE THEM WITH THE CALCIUM CHANCEL AND THE DOPA MINERGIC AND INSTEAD OF PRESENTING THE SECOND ORDER WITH A SUGAR REWARD, WE ACT VARMISIVATE THESE REWARDS AND ON POTENTIALLY THE REWARD WITH THE DOPA MINERGIC AND SEE WHAT HAPPENS. AND WE GOT THE ARE CULT, THE REASON I LABELED THE SCORES, WE GOT 90% OF THE FLIES ACTUALLY DOING THE RIGHT THING RATHER THAN THE TYPICAL 30%. WHICH IS WHEN WE ACTIVATE ABOUT 120 REWARD AND DOPA MINERGICOT SINGLE SIDE OF THE BRAIN WHEN WE ACTIVATE A SMALLER POPULATION, ABOUT 40 OR SO, WE GET A .5 SCORE WHICH IS 5% OF THE FLIES DOING THE RYE THING. I THINK THAT'S THE--THIS AMAIDSING PERFORMANCE REPRESENTS THE VAST EXPERIENCE OF FLIES EVER HAD IN ITS LIFE IT'S EQUIVALENT TO THE BEST 52 SLEEP AND DRINK AND SEX THAT IT EVER HAD AND 1 THING WE KNOW THIS IS BECAUSE IT'S A HETEROGENEOUS ROW GENIUS POPULATION OF NEURONS AND WE KNOW THE PERFORMANCE PERSISTS FOR ABOUT A WEEK AND INSENSITIVE AS TO WHETHER THE FLIES ARE FED OR STARVED AND WE KNOW THIS PARTICULAR PERFORMANCE IS ENTIRELY SATIABLE AFTER THE FOOD FROM THE TRAINING. SO THIS LED US TO THE IDEA THAT THE REWARDS THAT INNERIVATE THE TIP PROVIDES REINFORCES THE PROPERTIES OF SUGAR. NOW AS I SAID, THE SYNAPSE UNDER KNEES THESE NEURONS OR WHERE MEMORIES ARE LOCATED, AND ABLE TO FIND A SUBSET OF NEURONS FROM THE MUSHROOM BODY THAT HAVE A VERY LARGE DENDRITE IN THE TIP REGIONS OF THE MUSHROOM BODY AND HERE THERE'S--THERE'S ACTUALLY 3 THAT'S IN THIS HOLD OVER HERE AND YOU CAN SEE THEY HAVE A VERY DEFINED WELL DEFINED DENDRITIC CELL RIDDIC ARBOR ON A PARTICULAR PART OF THE HORIZONTAL TIP AND THESE TURN OUT TO BE DIFFERENT LOBES THAT I WON'T DETAIL. MUCH TODAY BUT WE'VE DONE A LOT OF WORK ON THIS MANY MONTHS AGO SO FINDING THESE LED US TO THE IDEA THAT IF THE SUGAR REWARD IS DELIVERED TO THESE STIPS THEN MAYBE REWARD CONDITIONING WILL CHANGE THE ORDER DRIVEN ACTIVITY OF THESE OUTPUT PATHWAYS. THAT WAS A SIMPLE EXPECTATION OF THE ANATOMICAL MODELS. AND SO WE'RE ABLE TO DO THIS OF COURSE, BY HEAD FIXING A TRAINED FLY AND THEN TESTING WHETHER THE ORDER OF ULTRA RESPONSE OF THESE OUTPUT NEURONS CHANGES BEFORE AND AFTER TRAINING. SO IN THIS KIND OF PREP, THEY SIMPLY PEEL OFF THESE CUPS AND WITH THE SALINE, STUCK SEE WHAT HAPPENED AND THIS SERIES OF EXPERIMENTS HE WAS ABLE TO SEE A FAIRLY SUBON THING THAT AFTER REWARD CONDITIONING, THE RELATIVE WOULD REVOKE RESPONSES OF THESE OUTPUT NEURONS APPEAR TO BE DECREASED. THIS IS A FAIRLY COMPLEX DISPLAY OF THE DATA, SUFFICE TO SAY IT LOOKS SOMEWHAT SIMILAR TO THE DEPRESSION THAT WAS FIRST SEEN IN THE LOCUST BECAUSE WE COULD TEST THIS IDEA. AND 1 OF THOSE THAT WAS SPECIFICALLY THE AWARD BY CONE DITIONING, I THOUGHT WE WOULD MIMIC IT BY BLOCKING THE OUTPUT BY A PATHWAY AND WE COULD DO THIS WITH A TOOL DEVELOPING BY TOSHI, SOME YEARS AGO SHOULD BE THE TEMPERATURE SENSITIVE DYNAMIC WE WOULD TAKE THE FLIES AND LURE THEM, AND AND 1 STRONGER STREAM AND AIR, AND THIS SCENARIO, THE DEFAULT CASES THAT THE FLIES AVOID A STRONGER STREAM. THE IDEA WAS IF THIS PARTICULAR SET OF OUTPUT NEURONS REPRESENTED THE NEURONS BY SWITCHING OFF THE NEURONS WAS ABLE TO AVOIDANCE WITH THE APPROACH. AND THAT'S CONDITIONS ON WHAT HAPPENED HERE AND THE NEXT GRAPH, IF THE BARS GO DOWN, THIS REPRESENTS AVOIDANCE, IF THE BARS GO UP, IT REPRESENTS APPROACH AND YOU CAN SEE THIS PARTICULAR ROTOCONCENTRATION, YOU SEE A STRONG BEHAVIORIAL AVOIDANCE OF .5 AND IF YOU SWITCH OFF THESE 2 OR 3 OUTPUT NEURONS ON THE BRAIN, THIS CONVERTS ON AUTODRIVEN APPROACH. NICE EVIDENCE THAT ACTUALLY MODIFYING THIS 1 PARTICULAR MUSHROOM BODY OUTPUT CONNECTION IS ABLE TO CHANGE THE BEHAVIOR AND ROTODRIVEN BEHAVIOR OF THE FLY FROM AVOIDANCE AND APPROACH AKIN TO TRAINING THEM WITH SUGAR REWARDS. SO THIS IS THE EVIDENCE THEN FOR THE MODEL I ALREADY PROVIDED YOU WITH. IN THE NAIVE SCENARIO, THE OVER ALL OUTPUT FROM THE MUSHROOM BODY, WERE SOMEWHAT BALANCED THEY WERE SOME THAT DRIVE AVOIDANCE, AND SOME THAT DRIVE APPROACH OR SIMPLICITY. AND OF COURSE, NEED TO BE BALANCED BY WEIGHT AND SO, THE NET BEHAVIOR DRIVEN BY THE MUSHROOM BODY IS A 0 AND THIS IS OUR SPACE, CONSISTENT WITH MANY, SOMEWHAT MISLEADING FINDINGS IN THE FIELD FROM LONG AGO THAT I CAN GO BACK THROUGH LATER THAT SUGGESTED THAT ACTUALLY THE MUSHROOM BODY WAS ONLY INVOLVED IN 1 BEHAVIOR BUT IT WAS SIMPLY BECAUSE--WHEN YOU BLOCK ALL OF THE MUSHROOM BODY, AND BLOCK ALL THE PATHWAYS AND APPROACH THE DERIVING PATHWAYS BUT WHEN YOU BLOCK SPECIFIC PATHWAYS, YOU SEE VERY RADICAL CHANGES IN WHAT LOOKS LIKE NAIVE ORDER DRIVEN BEHAIR AND THEN AS I SAID, AFTER LEARNING, YOU DECREASE THE SPECIFIC DRIVE TO SOME OF THESE AVOIDANCE PATHWAYS SO YOU SKEW THE WHOLE NETWORK, TOWARD DEVELOPING AND DRIVING APPROACH SO THIS IS SOMETHING THAT I'M GOING TO REVISIT AS I GO THROUGH THE TALK. THE IDEA THAT ACTUALLY THE OUTPUT NETWORK GIVES YOU THE VALENCE OF THE AUTODRIVEN BEHAVIOR, IN SOME CASES, JUST SKEW IT 1 WAY, YOU DRIVE THE APPROACH PATHWAYS AND IN OTHERS YOU CAN SPECIFICALLY DRIVE THE AVOIDANCE PATHWAYS. SO 1 THING THAT I WANT TO GO BACK TO IN MIKE'S DATA, IS THE FOLLOWING. SO HERE, ACTUALLY, IF NOT FAMILIAR WITH THE FIELD, YOU MIGHT THINK THAT ACTUALLY THE BEHAVIOR OF THE FLY THAT'S TRAINED FOR 2 MINUTES, CAN BE SEE AN A ROBUST APPROACH AND IT CAN BE SEEN A DAY LATER AND A DAY AND HALF LATER AND SO FORTH. HOWEVER THIS EXPERIMENT HAS NOT PROBE TRIALED KNOW ANIMALS. THIS IN EXPERIMENT, 1 POPULATION IS TRAINED AND THEN IT'S TESTED HALF AN HOUR LATER, IT'S COUNTED AND THROWN AWAY, A DIFFERENT POPULATION AND TRAINED, TESTED 12 HOURS LATER AND COUNTED AND THEN THROWN AWAY. THAT WAS ACTUALLY QUITE CONCONVENIENT TO GENERATE THIS DATA AND IT'S ALSO VERY INTERESTING BECAUSE IF YOU DO PROBE TRIAL THEM THIS IS THE BEHAVIOR. WHAT LOOKS LIKE A VERY STRONG MEMORY PERFORMANCE, WHEN TESTED A SECOND TIME AT 6 HOUR SYSTEM EFFECTIVELY 0. AND THIS WAS SOMETHING THAT JOHANNESS IN THE LAB TRIED TO UNDERSTAND. SO WHAT THIS REPRESENTED WAS DISTINCTION BY REWARD PREDICTION, IN THE FIELD, IT'S VERY PREVALENT WITH THE DOPA MINERGIC SIGNAL AND THE FLIES OF COURSE ARE TRAINED TO PREDICT THE PRESENTATION OF AN ORDER. THE MEANS FOR THIS, AND 1 MIGHT ASSUME THEN THAT IF THEY GO INTO THAT ORDER AND FOOD IS NOT THERE, THEN THEY WOULD REEVALUATE WHAT THIS MEANS FOR THEM AND THEN CHANGE THE SUBSET WENT BEHAVIOR AND SO JOHANNES, SHOWED THIS NICELY IN THE SERIES OF EXPERIMENT. SO THE FLIES ARE TRAINED, 1 ORDER WITHOUT REWARD, 1 WITH AND THEN 3 HOURS LATER HE REACTIVATES THE MEMORY BY GIVING HIM EITHER THE PREVIOUSLY UNREWARDED ORDER OR THE PREVIOUSLY REWARDED ORDER OR NOVEL ORDER OR AIR AND HE GIVES THEM ANOTHER 3 HOURS, HE DOES THIS TWICE, THIS 3 OUT POSITION AND THEN HE GIVES THEM ANOTHER 3 HOURS AND HE TESTS THE PERFORMANCE AND AS YOU CAN SEE HERE, IF THERE THEY HAD BEEN PREEXPOSED TO THE FLUS ORDER, 3 HOURS AFTER TRAINING, THE 6 HOUR MEMORY PERFORMANCE IS COMPLETELY ABSENT WHERE IF THEY WERE GIVEN THE PREVIOUSLY UNREWARDED ORDER THERE'S NO CONSEQUENCE, LIKE WISE AIR AND THERE'S NO CONSEQUENCE SO THERE THIS IS VERY GOOD EVIDENCE FOR CONDITION STIMULUS DEPENDENT PREVIOUSLY REWARDED DEPEBBLED END MEMORY EXTENSION. AS I SAID THE MODEL IS THAT REWARD LEARNING SKEWS THE NETWORK. THAT REWARD LEARNING SPECIFICALLY DECREASES THE AUTODRIVE TO THESE AVOIDANCE DIRECTING MUSHROOM BODY OUTPUT NEURONS SO WE ASSUME THAT THE DISTINCTION PROCESS WILL BE A PROPERTY THAT THIS INPUT NETWORK THAT WAS DURING LANDING AND THE REASON FOR THAT IS THAT THERE'S A LOT OF EVIDENCE TO SUGGEST THAT MOST OF THESE OR MANY OF THESE OUTPUT NEURON PATHWAYS ARE ACTUALLY RECOMBIN ANTLY CONNECTED TO THOSE THAT INNERIVATE THE SAME ZONE. SO IF YOU LIKE EACH 1 OF THESE ZONES IT'S ITS OWN LITTLE DOPA MINERGIC FEEDBACK, THE POTENTIAL TO REEVALUATE THE PLASTICITY THAT'S BEEN IMPOSED ON ITS OWN ZONE. SO THE FUNCTION WAS THEN THAT IF REWARD LEARNING ACTUALLY DECREASED THE DRIVE TO THE AX VOIDANCE PATHWAY WHEN THE FLY WAS GIVEN THE REWARDED ORDER FOR THE SECOND TIME, IT WOULD PREVENTIVE RENTERALLY ACTIVATE THE REINFORCING PATHWAYS DRIVEN BY THE APPROACH DIRECTING MUSHROOM BODY OUTPUT NEURONS. SO JOHANNES, TESTED THIS MODEL USING AGAIN, SOME OF THESE TO FIND THE RIGHT LINES DEVELOPED ON THE SPLIT GULF OR TECHNOLOGY. HERE'S 1 THAT LABELS VERY SPECIFICALLY A SUBSET OF MUSHROOM BODY OUTPUT NEURONSOT VERTICAL LOBES OF THE MUSHROOM BODY WHERE LOTS OF NAMES ARE LARGELY MEANINGLESS TO MYSELF INCLUDED IN THE 1--I'LL TRY NOT TO USE. BUT THESE ONES ON THE VERTICAL OUTPUT NEURONS CAN BE LABELED VERY NICELY WITH THIS DRIVER AND SO JOHANNESS ASKED WHAT HAPPENS TO MEMORY EXTINCTION IF I BLOCK THIS OUTPUT PATHWAY WITH USA H-SHI, AND HE HE BLOCK ITS WITH THE MEMORY TRAINING, IT'S NOT CLEAR. SUGGESTING THAT ACTUALLY THE OUTPUT FROM THE VERTICAL PROJECTION THAT COULD POTENTIALLY DRIVE REINFORCING NEURON SYSTEM REQUIRED FOR THIS EXTINCTION PROCESS AND NOTHING CLEAR APPEARS TO HAPPEN WITH THE PREVIOUSLY REINFORCED ORDER SO, FIRST GOOD SIGN THAT MAYBE THE VERTICAL OUTPUT NEURONS OF THE THING THAT'S DRIVING THE EXTENSION PROCESS. NOW AS I SAID TO YOU, THE ASSUMPTION WAS THAT THESE MIGHT FEEDBACK ON TO ENFORCING NEURONS THAT INNERIVATE THE SAME ZONE. SO THE NEXT THING HE DID WAS JUST WHO SHOULD BE IN THESE REINFORCING DOPA MINERGIC NEUROWNS THAT INNERIVATE THE SAME ZONES AND SWITCH THEM OFF DURING THE SECOND ORDER, REACTIVATION PHASE AND SEE WHAT HAPPENS TO THE EXTINCTION AND YOU CAN SEE VERY NICELY THAT ACTUALLY THE CS + NO LONGER DRIVES ROBUST EXTENSION. THE OTHER THING THAT EMERGES IF YOU'RE PAYING ATTENTION HERE IS THAT WE NOW SEE A VERY PRONOUNCED EFFECT WITH THE CS MINUS. THIS WAS NOT EXPECTED BEFORE, WE BLOCKED ANY DOPA MINERGIC NEURONS, IF YOU REACTIVATED THE MEMORY WITH THE PREVIOUSLY NONREINFORCED ORDER, DON'T DO ANYTHING AT ALL BUT HE AND NOW, THE MEMORY PERFORMANCE IS GONE AND I'LL COME BACK TO THIS SOMEWHAT LATER. BUT THIS FINDING WITH THE CSPLUS IS HIGHLY CONSISTENT WITH THE IDEA THAT THE OUTPUT NEURONSOT VERTICAL LOBE DRIVE THE DOPA MINERGIC NEURONS THAT FEEDBACK ON TO THE VERTICAL LOBE, SO OF COURSE WE CAN TEST THIS KIND OF FUNCTIONAL CONNECTIVITY USING THE DIFFERENT TOOLS THAT ARE AVAILABLE TO GENERATE TISSUE SPECIFIC EXPRESSION, SPECIFICALLY LET'S SAY, THE CONTROL IN [INDISCERNIBLE] 4, AND FOR EXAMPLE, IF WE PUT 1 OF THE CHANNEL REDOBSIN VARIANTS IN THE VERTICAL LOBE OUTPUT NEURONS AND WE PUTTED GCAMP IN THE NEURONS WE CAN STIMULATE THE OUTPUT NEURONS AND SIMPLY LOOK AND SEE, IF WE SEE ANY RESPONSES, ANY EVIDENCE OF FUNCTIONAL CONNECTIVITY IN THE DOPA MINERGIC NEURONS AND THESE EXPERIMENTS WE'RE ABLE TO SEE FAIRLYBUT CONSISTENT RESPONSES IN SOME OF THE CELLS, THIS 1 SEEMS TO BE ACTIVATED, THIS 1 SEEMS TO BE INHIBITED AND THIS 1 AT LEAST THE SAMPLE NUMBER DOESN'T APPEAR TO BE DOING A GREAT DEAL. SO THIS SUGGESTEDS THAT THERE'S SOME FUNCTIONAL CONNECTIVITY BETWEEN THESE OUTPUT NEURONS AND THESE DOPA MINERGIC NEURONS CONSIST WENT THE IDEA THAT AFTER REWARD TRAINING, THE FLY DRIVES THIS PATHWAY WITH AUTOREACTIVATION WHICH ACTIVATES THESE RECURRENT PATHWAYS CONSISTENT WITH THE IDEA THAT THIS IS WHAT HAPPENS DURING EXTENSION, YOU SPECIFICALLY DRIVE THIS ACTIVITY AND THE REINFORCING DOPA MINERGIC ARM AND THE MODEL OF COURSE IS THAT THE EXTINCTION S&P THAT THE FORMATION OF A COMPETING MEMORY WHICH EFFECTIVELY BLOCKS THE AUTODRIVE TO SOME OF THESE APPROACH PATHWAYS SO IT RETURNS THE FLY TO PSEUDOKIND OF NAIVE STATE WHERE THE NETWORK IS BALANCED AS IT WAS BEFORE IT WAS EVER TRAINED. OF COURSE THERE WERE SOME ASSUMPTIONS AND 1 OF THE INTERESTING THINGS IS THIS PHENOMENON WHERE OVER TIME, THE MEMORY PERFORMS REAPPEARS AND WE HAVEN'T SEEN THE EVIDENCE FOR THAT BUT I THINK EVERYTHING I'VE SHOWN YOU IS CONSISTENT WITH THE IDEA THAT THE REINFORCING PATHWAYS TRY TO FORMATION OF THE MEMORY AND THIS IS ACTUALLY HOW EXTENSION IS MANIFESTED IN THE NERVOUS SYSTEM. NOW, 1 THING THAT I'VE ALREADY ALLUDED TO IS THAT 1 OF THE THICKS THAT EMERGE WHEN WE BLOCK THE REINFORCING NEURON SYSTEM THAT WE UNCOVERED THIS EFFECT WITH A CS-MINUS KEY. JUST TO REITERATE IF WE BLOCKED ALL OF THESE REINFORCING NEURONS WE INHIBIT THE CS + DRIVEN EXTENSION BUT WE ALSO SEE THAT THE CS MINUS NOW DECREASES THE APPROACH BEHAVIOR OBSERVED EVERY 6 HOURS AND THIS MAY VERY LITTLE SENSE TO US OTHER THAN IF WE CONSIDER THIS. WE WONDERED WHETHER OR NOT WHAT WE WERE LOOKING AT HERE WAS A MEMORY RECONSOLIDATION TYPE OF PHENOMENON. NOW I'M ASSUMING MANY OF YOU KNOW WHAT MEMORY RECONSOLIDATION IS BUT IT WAS REINVIGORATED 10-12 YEARS AGO, MAYBE 10 YEARS AGO BY KOREAN NATIVE'S LAB, THE IDEA BEING THAT WHEN MEMORY CONSOLIDATED MEMORY IS RETRIEVED IT RETURNS TO A FAIRLY LABEL STAGE THAT MAY IN SOME RESPECTS REPRESENT A SHORT-TERM MEMORY. THIS IS OF COURSE RECEIVED A HUGE AMOUNT OF INTEREST BECAUSE IT'S USEFUL FOR EXAMPLE CURE POST-TRAUMATIC STRESS DISORDER IN PEOPLE RETURN FREE RADICALS GENERATED WAR, MAYBE PTSD IN RESPONSE TO PRESIDENTIAL ELECTIONS, AND SO FORTH. THE IDEA BEING THAT WHEN YOU RETRIEVE MEMORY PERFORMANCE AND FOR EXAMPLE, MINISTER THE DRUG PROPONANOLE, YOU REVERSE THESE MEMORIES SOME PEOPLE HAVE HINT THAD THIS COULD BE USED TO REMOVE DRUG-SEEKING MEMORIES. THIS IS GENERAL INTEREST AND IT'S SOMETHING I LOOKED UP MANY, MANY YEARS AGO IN THE FLY AND SO WE'RE EXCITED THIS MIGHT BE A SOLID ENTRY POINT AND TO TRY TO UNDERSTAND WHAT MEMORY RECONSOLIDATION IS, IT'S HEAVILY STUDY TODAYED EYE A NUMBER OF LABS AND I DON'T THINK WE REALLY KNOW WHAT'S GOING ON. SO THIS OF COURSE WAS THE GRAPH THAT I SHOWED YOU EARLY EARLY FROM MIKE. THIS IS THE EVIDENCE FOR THE CONSOLIDATION OF FLY MEMORY THAT I SAID BY 2 HOURS LEFT, AFTER TRAINING THE MEMORY IS RESISTANT TO COLD SHOCK, AND SUGGESTING IT'S OPERATIONALLY CONSOLIDATED AND THE GUESS WAS THAT IF WHAT WE WERE LOOKING AT WAS MEMORY RECONSOLIDATION, THEN IT SHOULD RETURN LATE OR WHEN REACTIVATED BY THE PREVIOUSLY NONREINFORCED ORD TORE A STATE THAT WAS AGAIN DISRUPTIBLE BY COLD SHOCK. SO THAT'S THE EXPERIMENT THAT JOHANNESS DID. THIS PROTOCOL WILL COME UP FOR THE REST OF THE TALK. SO I WANT TO MAKE IT CLEAR. SO HE TRAINS THE FLIES, 1 ORDER, OR ANOTHER WITH REWARD AND THEN 3 HOURS LATER HE SPECIFICALLY REACTIVATES THE MEMORY WITH THE PREVIOUSLY NONREWARDED ORDER. TO REITERATE THE PREVIOUSLY REWARDED ORDER, CSPLUGS GAVE US STRONG MEMORY EXTENSION AND THE CS MINUS DIDN'T DO ANYTHING UNLESS WE BLOCK THESE NEURONS. HE THEN FOLLOWED THE REACTIVATION OF THE MEMORY WITH PLUMPLEGGING THE FLIES INTO THE ICE BUCKET TO SEE WHETHER THE MEMORY RETURNED TO LABEL AND THEN HE GAVE THEM THE 3 HOURS TO RECOVER AND HE MEASURED 6 HOUR MEMORY PERFORMANCE. AND HOPEFULLY YOU CAN SEE HERE THAT IF HE GIVES THE FLIES NO TREATMENT AT ALL, HE HAS THIS KIND OF PERFORMANCE IF HE GIVES THEM ONLY A COLD SHOCK SO HE DOESN'T REACTIVATE THE MEMORY, YOU CAN SEE THE MEMORY IS IN THE CONSOLIDATED STATE AND IF HE ONLY GIVES HIM THE CSMINUS REACTIVATION BUT NO COLD SHOCK, YOU GET STRONG PERFORMANCE BUT HE COME BINES THE CS MINUS REACTIVATION OF THE MEMORY AND QUICKLY FOLLOWS IT, WITH COLD SHOCK, THE MEMORY PERFORMANCE IS GONE. I THINK REALLY NICE EVIDENCE THAT ACTUALLY CS MINUS DRIVEN REACTIVATION DRIVES MEMORY RECONSOLIDATION PROCESS, WHERE I SAID, THE CS + PROCESS DRIVES THE REEVALUATION OF PROCESS OF EXTINCTION. SO I THINK FOR THE FIRST TIME NICELY DIFFERENTIATING BETWEEN THE PROCESS OF EXTENSION AND RECONSOLIDATION SIMPLY WITH STIMULUS PRESENTATION. THE OTHER THING THAT HE SHOWED WAS THE RECONSOLIDATION PHENOMENON LIKE CONSOLIDATION IS TIME DEPENDENT PROCESS AS YOU MIGHT EXPECT AND IF HE FOLLOWS THE REACTIVATION WITH AN IMMEDIATE COLD SHOCK OR 30 MINUTES LATER, HE SEES MEMORY IS LOST BUT HE WAITS AN HOUR AND A HALF, IT'S RECONSOLIDATED, IT'S AGAIN INSENSITIVE TO THIS KIND OF DISREPRESENTATION. SO OF COURSE, WE WANTED TO KNOW WHAT ARE THE PROCESSES, WHAT ARE THE NEURONS THAT ARE DRIVING THIS PHENOMENON AND WE ALREADY SOME SOME INSIGHT, THOSE--THIS DOPA MINERGIC NEURON BLOCK HAD REVEALED THAT SOME OF THESE REINFORCING NEURONS WERE INVOLVED AND OF COURSE WE HAD SOME EXPECTATIONS FOR WHAT WE WOULD EXPECT TO SEE IN THE RECONSOLIDATION CIRCUIT. THE FIRST WAS THAT THEY MUST SOMEHOW REPRESENT THE NONREWARDED ORDER. THE BIAS FROM THE FIELD FOR MANY YEARS HAS BEEN THAT AFTER CONDITIONING WITH SUGAR AND ODOR, ONLY IT'S REWARRED IN THE NERVOUS SYSTEM, THE CS-IS COMPLETELY IRRELEVANT. THIS SHOWED YOU THAT THAT'S COMPLETELY WRONG BECAUSE THE CS- SPECIFICALLY DRIVEWAYS THE RECONSOLIDATION PROCESS SO IT HAS TO BE REPRESENTED SOMEWHERE IN THE NETWORK. SO WE NEEDED TO BE ABLE TO FIND WHERE IT WAS REPRESENTED IN THE NETWORK. THE OTHER THING IS THAT THE RECONSOLIDATION PHENOMENON SUGGESTS THAT SOMETHING HAPPENS WITH THE ORIGINAL REWARD MEMORY AND SOMETHING HAPPENS TO THE SUGAR REWARD ORDER REPRESENTATION SO WE NEEDED TO SOMEHOW ACCOMMODATE THAT IN THE MODEL. SO, THIS WAS THE DATA I'VE ALREADY SHOWN YOU, IF JOHANNESS FOLLOWED THE REACTIVATION OF THE CS- AND HE BLOCKED ALL OF THESE NEURONS, HE BLOCKS EXTINCTION, HE BLOCKED THE CS- WHERE IT DRIVES THE MEMORY LOSS AND THE ASSUMPTION WAS THAT THE ACTIVITY OF THESE NEURONS WAS REQUIRED FOR THE RECONSOLIDATION PROCESS AND SO IF YOU BLOCK THEM, THE MEMORY DISAPPEARS. SO OF COURSE THIS IS LIKE 4 OR 5 SUBSETS OF REWARDING THE NEURONS, THERE ARE TOOLS TO LOOK AT THEM, AND THAT'S WHATEE DID, HE FIRST USED INDIVIDUAL TOOLS TO ACTIVATE SPECIFIC SUBSETS OF ALL THE VERTICALLY PROJECTING AND DOPA MINERGIC NEURONS AND HE FOUND THE SPECIFIC SUBSET THAT INENERIVATES THIS BLOG WHICH IS CALLED MV-1 GIVES HAD IMPEDIMENTS A VERY NICE LOSS OF PERFORMANCE AFTER REACTIVATION. SO IT'S THIS 1 SPECIFIC SUBSET THAT INNERIVATES THIS ZONE THAT'S INVOLVED, THE OTHER THING I WANT TO POINT OUT IS THAT THE EXTINCTION PHENOMENON IS LESS THAN INTACT, THE CS+ STILL DRIVES EXTENSION BUT NOW WHEN WE BLOCK THIS NEURON, THE CS- NOW DRIVES COMPLETE MEMORY LOSS. SO THE IDEA BEING THIS PARTICULAR DOPA MINERGIC PATHWAY WAS MAYBE SPECIFICALLY INVOLVED IN DRIVING CONSOLIDATION PHENOMENON. ONE OF THE THINGS THAT I GUESS I BELABORED IS THAT THE SPECIFIC FOR ZONES, THESE ZONES HAVE A SUBSET NEURON WITH THE DENDRITIC CELL RIDE IN THE SAME ZONE, THIS IS THE MIEWSH ROOM BODY OUTPUT NEURON THAT HAS THE DENDRITE IN THE SAME ZONE AS THE KEY RECONSOLIDATION SPECIFIC DOPA MINERGIC NEURON AND SO WE ASSUMED THIS PARTICULAR ZONE IN THIS PARTICULAR RECOMBIN ANT NETWORK MAY BE INVOLVE WIDE DRIVING THE ENTIRE RECONSOLIDATION PHENOMENON. SO JOHANNESSICAL EVERYONED THIS AGAIN TO BLOCK THESE NEURONS AND ASK WHETHER THEY WERE INVOLVED IN THE RECONSOLIDATION PHENOMENON. AND HERE'S THE KEY EXPERIMENT IF HE FOLLOWS THE--IF HE SORT OF BLOCKS THIS OUTPUT PATHWAY DURING--SPECIFICALLY DURING MEMORY REACTIVATION, THEN ALLOWS IT TO REGAIN FUNCTION BY REDUCING THE TEMPERATURE AND THE 6-HOUR MEMORY, YOU CAN SEE THE EXTENSION STILL HAPPENS, AND HE HOW SEES THIS FAIRLY STRONG RESPONSE ON THE RECONSOLIDATION PHENOMENON. CONSISTENT WITH THE IDEA THAT THIS LOCAL RECURRENCE MIGHT ACTUALLY BE INVOLVED. THE OTHER THING WE CAME TO SEE WAS THE TO SEE THEM IN THE ORDER. SO IN THESE EXPERIMENTS WE DID THE ORDER BEFORE AND AFTER TRAINING AND WE CHALLENGED THE ACTIVITY IN THIS PARTICULAR OUTPUT NEURON WITH BOTH THE CS+ AND THE CS- ORDER AND ASKED WHETHER ANYTHING CHANGED. AND 1 OF OF THE THINGS THAT EMERGED THAT SURPRISED US A GREAT DEAL WAS THAT IF WE TRAIN THE FLIES WITH METHYL AND SUGAR WE SEE A SPECIFIC INCREASE IN THE RESPONSE DRIVEN BY OCT PREVIOUSLY SHOWN QUAWBTIFIED HERE IF WE TRAIN THE FLIES WITH OCT IN THE SUGAR WE SEE A SPECIFIC INCREASE IN THE RESPONSE GENERATED BY THE PREVIOUSLY NONREINFORCED METHYL CH. I THINK THIS IS BEAUTIFUL COMPARED TO MOCK. AND TO OUR GREAT SURPRISE AND I SHOULD SAY WE DON'T ACTUALLY NOW KNOW THIS COMES AROUND BUT WE DO SEE A SPECIFIC INCREASE IN THE PREVIOUSLY NONORDERRED DRIVE TO THE PARTICULAR MUSHROOM BODY OUTPUT NEURON PATHWAY. SO THAT'S GOOD. THE OUTPUT NEURONS ARE REQUIRED AND THE DOPA MINERGIC NEURONS ARE REQUIRED AND WE SEEM TO HAVE GAINED THE SPECIFIC INCREASE IN ORDER DRIVES TO THIS COMPARTMENT BUT OF COURSE, WE WANT TO KNOW WHETHER THESE ARE ACTUALLY FUNCTIONALLY CONNECTED SO AGAIN, WE DO THE FUNCTIONAL CONNECTIVITY EXPERIMENTS, I SHOULD SAY THESE ARE ALL IN AN EX-PLANT PREPARATION AND IT'S FOR THE COVER SLIP, ALL OVER [INDISCERNIBLE] SAID EXPRESSED CRIMSON IN THIS PARTICULAR OUTPUT NEURON PATHWAY IN G-CAMP IN THE DOPA MINERGIC NEURONS THAT PROMOTIONAL RECURRENT AND HERE WE SEE A INEDIBLY ROBUST CONNECTIVITY WHEN ACTIVATING THE ANTIBODY WE SEE IGF 95 CANT INCREASE IN RESPONSES IN THIS PATHWAY, CONSISTENT WITH THIS IDEA THAT THE RECURRENCY RESULTS. ONE OF THE COOY THINGS THAT DRIVES THE MEMORY RECONSOLIDATION PHENOMENON, DURING MEMORY REACTIVATION WITH THE MINUS. SO THE SECOND THING I SAID WE HAD TO FIND WAS SOME SORT OF EVIDENCE THAT THIS COULD BE DOING SOMETHING FOR THE PREVIOUS REWARDED MEMORY SO THE FIRST THING THAT JOHANNES DID WAS COME UP WITH THIS CLEVER EXPERIMENT, I THINK SO HE TRAINED THE FLIES WITH 1 ORDER OF NO SUGAR, 1 WITHOUT AND THEN HE REACTIVATED THE MEMORY WITH THE PREVIOUSLY NONWRITTEN SEED AND THEN HE TESTED THE FLIES FOR THE CHOICE BETWEEN THE PREVIOUSLY AWARDED ORDER AND 1 THEY HAD NEVER SEEN BEFORE: THE IDEA WOULD BE OF COURSE THAT IF THEY STILL HAD A STRONG REWARD MEMORY FOR THE SUGAR REINFORCED ORDER THEN THEY SHOULD APPROACH IT. IF REACTIVATION WAS A PREVIOUSLY NONREINFORCED ORDER, DID SOMETHING TO THE PREVIOUSLY REWARDED ORDER MEMORY, THEN IT WOULD PRESUMABLY NOT KNOW WHAT TO DO WHEN GIB THIS CHOICE OF CS+ VERSUS NOVEL, AND AS CAN YOU SEE HERE, VERY SIGNATURES 95 KACCT TININE PATHWAYLY, THE MEMORY IS LOST BOY THE PRESENTATION WITH THE PREVIOUSLY NONREWARDED OR 3 OURS BEFORE HAND. SO I THINK VERY NICE EVIDENCE THAT ACTUALLY THE CS MINUS AND THE CS + ARE FUNCTIONALLY CONNECT INDEED THE NERVOUS SYSTEM SOMEHOW. THAT THESE MEMORIES ARE ACTUALLY REPRESENTED SOMEWHAT TOGETHER ALTHOUGH WE THINK PROMOTIONAL IN DIFFERENT PLACES WITHIN THE ENSEMBLE AND SOMEHOW OR ANOTHER AND I WOULD SAY WE DON'T UNDERSTAND HOW, THESE CONNECTIONS ACTUALLY OF THE MINUS REACTIVATION VIA THIS KIND OF ZONE IMPACTS THE REWARD MEMORY THAT'SOT HORIZONTAL TAPE. I THINK I SHOWED YOU, SOME OF THE EVIDENCE THAT WE KNOW THAT THE REWARDED MEMORY IS IN THE HORIZONTAL TIP AND WE WANT TO BE ABLE TO SOMEHOW CONNECT THESE 2 DIFFERENT ZONES AND THESE CS- CS+ MEMORIES. ONE OF THE FIRST THINGS TO DO WAS TO SEE WHETHER THIS OUTPUT PATHWAY, FROM THE ZONE IN BLUE HERE ACTUALLY COULD POTENTIALLY FUNCTIONALLY CONNECT TO THE REWARD IN DOPA MINERGIC NEURONS AND SO THE IDEA BEING THAT IN ADDITION TO DRIVING RECURRENCE WITHIN THIS LOCATION WITH THE CS- PARADIGM OF THE MEMORY TICKETED DO SOMETHING TO THE PREVIOUS ISSUE, THE REWARDED MEMORY THAT I TOLD YOU WAS ON THE HORIZONTA TIP. AND WHEN WE DO THIS FUNCTIONAL CONNECTIVITY EXPERIMENT AGAIN WITH CRIMSON IN OUTPUT PATHWAY, AND REWARDING DOPA MINERGIC INUREONS YOU CAN AGAIN SEE FAIRLY STRONG EVIDENCE OR FUNCTIONAL CONNECTIVITY BETWEEN OUTPUT NEURON PATHWAY THAT'S IN THIS ZONE AND DOPA MINERGIC NEURONS THAT INNOVATE THE TIPS THAT ARE INVOLVED IN SUGAR REWARD THE ENFORCEMENT SO POTENTIALLY SOMETHING WAS HAPPENING BETWEEN THE CSMINUS REACTIVATED PATHWAY AND THE REWARD IN DOPA MINERGIC NEURONS THAT WERE FORM INDEED FORMING THE SUGAR SOABTED ORDER MEMORY. THE KEY EXPERIMENT WAS THIS 1. THE ASSUMPTION WAS THAT IF THESE REWARD MEMORIES, THE REWARD MEMORY REINFORCING DOPA MINERGIC NEURONS WERE REQUIRED AGAIN, THEN WE SHOULD FURTHER IMPAIR FUNCTION AFTER MEMORY REACTIVATION WITH THE CS- AND WE WOULD THEN SEE THE MEMORY DISAPPEARED AND ACTUALLY THAT'S EXACTLY HAPPENED. IF WE TRAIN THE FLIES, 1 ORDER, NO SUGAR REWARD, 1 WITH, AND THEN 3 HOURS LATER, CHALLENGE THEM WITH THE PREVIOUSLY UNREINFORCED ORDER TO TRIGGER MEMORY RECONSOLIDATION AND THEN IMMEDIATELY AFTER THE ORDER EXPOSURE, WE BLOCK THE ACTIVITY OF THESE REWARD AND DOPA MINERGIC NEURONS, YOU SEE THE PERFORMANCE IS GONE AND CONSISTENT WITH THE PREVIOUS EXPERIMENT THAT SHOW THAT THE RECONSOLIDATION PROCESS IS TIME DEPENDENT IF WE WAIT FOR, I THINK AN HOUR AND A HALF HERE AND BLOCK THE PATHWAY AGAIN, AND YOU SEE THAT THE MEMORY IS MAINTAINED. SO JUST LIKE THE COLD SHOCK EXPERIMENT BEFORE, THE ABILITY OF THE MEMORY IS TIME DEPENDENT WITH THE BLOCKADE OF THE REWARD AND DOPA MINERGIC NEURONS. I THINK QUITE NICELY SHOWING THAT ACTUALLY THE SECOND PART OF THE RE CONSOLIDATION PHENOMENON IS DRIB BY THE UT PUT PATHWAY REENGAGING THE REWARDING POPULATION OF DOPA MINERGIC NEURONS THAT PRESUMABLY RECONSOLIDATE THE PROCESS THAT WAS ORIGINALLY FORMED DURING MEMORY. SO THAT'S EFFECTIVELY ALL I WANT TO SAY TODAY. I WANT TO LEAVE YOU WITH THE GENERAL IDEA THAT ACTUALLY, MUCH OF THE MEMORY FORMATION AND REEVALUATION KIND OF PHENOMENA ARE MEDIATED BY THE HEAVILY INTERCONNECTED OUTPUT NETWORK BETWEEN THE 5000 OR SO MUSHROOM BODY CONNION CELLS AND A HANDFUL OF OUTPUT PATHWAYS THAT ARE THAT FEEDBACK MOST OF WHICH FEEDBACK INTO DOPA MINERGIC NEURONS WITHIN THE SAME ZONE PROVIDING THIS VALUE, IT'S KIND OF FUNCTION AND I SHOWED YOU THAT IN--IN THE CASE OF EXTENSION, YOU CAN DRIVE THIS TO FORM A COMPETING AVERSIVE MEMORY AND RECONSOLIDATION DRIVEN BY THE PREVIOUSLY NONREWARDED ORDER, YOU DRIVE THIS RECOMBIN ANT PATHWAY THAT'S SPECIFIC TO THIS KIND OF ZONE IN THE CORNER IN ADDITION AND DELAYED MANNER, YOU DRIVE THE PREVIOUSLY LEARNING SPECIFIC REINFORCING NEURONS TO RECONSOLIDATE THE ORIGINAL REWARD MEMORY. I THINK 1 OF THOSE THINGS TO SAY IS THAT 1 OF THE THINGS THAT'S EMERGE FRIDAY THIS KIND OF STUDY IS THAT THE IDEA THAT THERE'S ACTUALLY A LARGE ERROR DURING MEMORY TESTING, REWARD, NO SUGAR PRESENT DRIVES DISTINCTION AND SMALL ODOR, BECAUSE HALF OF THE TRAINING TRIAL, THEY RECEIVE THE CS- WITH NO SUGAR AND THEN IN THE TRAINING THEY RECEIVE THE CS+ WITH THE SUGAR AND THIS REACTIVATION WE DON'T GET IN THE SECOND ORDER OF THE SUGAR. WE ASSUME THAT DRIVES THE CONSOLIDATION SO THIS MAY BE AN ERROR THAT DRIVES DISTINCTION VERSE US CONSOLIDATION AND THAT'S SOMETHING WE WILL LOOK AT MORE REGULARLY. SO JUST TO FINISH THEN. I WANT TO REACKNOWLEDGE THE PEOPLE THAT DID ALL THE WORK ON THE RECONSOLIDATION AND EXTENSION, PROJECT, JOHANNESS LED WITH VERY NICE HELP FROM KD--SALLY AND ALL OF THE FUNCTIONAL CONNECTIVITY IS IMAGING WAS DONE BY OLIVER BARNSTEDT, WE ARE ABLY FUNDED BY A NICE GROUP OF FUNDERS IN A NICE PLACE AND WITH THAT I'LL TAKE PLACE. [ APPLAUSE ] >> SO THERE'S 1 OR 2 THINGS I'M STRUGGLING WITH, I'M STRUGGLING SO ALL OF THE THESE ARE AVAILABLE AT THE LEVEL OF INTERACTION OF DOPA MINE NEURONS WITH THE OUTPUT NEURONS HOW DO YOU GET DIFFERENT VALENCES TO DIFFERENT STIMULI, RIGHT? IF YOU'RE TALKING NOW NOT ABOUT A CS AND CSPLUS AND MINUS GIVEN AT THE SAME TIME BUT A CS WAS GIVEN 2 HOURS BEFORE OR A DAY BEFORE, HOW DOES THE SLIDE TELL TELL THE DIFFERENCE AND DO THE PAIRING AT THE SAME TIME AND NOT HAVE THE OUTPUT NETWORK APPLIED TO ALL THE OTHER MEMORIES THAT'S GOT STORED UP? >> SO IF I UNDERSTAND THE QUESTION, SO-- >> I MAY NOT BE SAYING IT CORRECTLY. >> I THINK IT'S A GOOD QUESTION. SO WHAT YOU'RE SAYING IS IF THE VALANCE AWARD SUGAR MEMORY IS REPRESENTED AS INHIBITION ON THIS PARTICULAR PATHWAYOT TIP, HOW DOES THAT NOT HAVE AN EFFECT ON A WATER MEMORY OR SO FORTH? WELL WE KNOW THAT AND WE LOOKED INTO THIS, I DIDN'T TALK ABOUT IT THEA ALL TODAY BUT 1 OF THE OTHER STRAPPEDS OF WORK THE LAB HAS DONE IS TO LOOK AT DIFFERENT REINFORCERS SO REWARDING DIFFERENT REINFORCERS AND THE THING THAT WAS EASY TO DO IS FIRST REWARD WITH WATER AND THE WORK SUGGEST SAYS THAT ACTUALLY, WHEREAS THE SUGAR REINFORCEMENT IS IN THE END BLOG, THE WATER REINFORCEMENT IS THE NEIGHBORING ZONE. SO THE VALANCE IS THE SAME BECAUSE IT'S AFTER AN INHIBITING DRIVE TO THE AVOIDANCE PATHWAYS BUT THE SPECIFICITY OF THE REINFORCEMENT IS ZONE SPECIFIC AND 1 OTHER INTERESTING THEN IS THAT THE STATE DEPENDENCE APPEARS TO BE INDEPENDENT SO THE STATE DEPENDENCE AND THIS IS SOMETHING THAT MIKE FIRST LOOKED AT IN THE LAB AND RELAYED FOR HUNGER ON THIS ZONE HERE,OT TIP AND VIA FEET FORWARD INHIBITORY PATHWAY SEEMS TO SPECIFICALLY EFFECT THIS TERMINAL ZONE. >> BUT LET'S SAY-- >> DOESN'T EFFECT THE FIRST PART. >> BUT LET'S SAY YOU HAD A SUGAR REWARD PAIRED TO OCTHIS MORNING AND NOW YOU USE SUGAR AS A REWARD FOR BANANA THIS AFTERNOON, IS IT GOING TO BE ABLE TO TELL THOSE APART IF YOU TEST THEM TONIGHT? OR BECAUSE NOW YOU'RE GOING--GETTING THE SAME OUTPUT. >> OF COURSE IT'S AN ODOR SPECIFIC CANYON CELL TO THE OUTPUT NEURON BUT WE'RE BOTTLENECKING ON THE SAME OUTPUT SO THAT'S A GOOD QUESTION. SO THE ASSUMPTION WOULD BE THAT OF COURSE IT'S VALENCE AND THERE'S TIME DEPENDENT TO THIS, THE PERFORMANCE IS STRONGERERABILITIER THAN LADIER, IF YOU WERE ABLE TO DO THEM AT THE SAME TIME, I ASSUME THEY WOULD SIT BETWEEN THE 2 ORDERS AND LIKE THEM BOTH. >> I DON'T THINK YOU WOULD EXPECT ANYTHING ELSE. >> THE QUESTION OF RELATIVE KIND OF VALUE, WE LOOKED AT IN SOME RESPECTS BECAUSE THE SUGAR THAT JUST TASTES TWEET VERSUS 1 THAT'S SWEET AND NUTRITIOUS, THEY WILL REFER SUITE AND NUTRITION AND I THINK THAT ACTUALLY AGAIN COMES ABOUT I HAVE DIFFERENT [INDISCERNIBLE]. >> SURE, BUT THE SWEET REINFORCEMENTS DOWN HERE, THIS LITTLE GREEN BLOB THAT'S HITTEN SOMEWHAT IN THIS DIAGRAM APPEARS TO BE THE NUTRIENT REINFORCER SIGNAL THERE SO THEY'RE SIMPLY A DIFFERENCE IN THE NUMBER OF OUTPUT PATHWAYS THAT ARE CHANGED BY THE DIFFERENT REINFORCERS. SO I'M NOT ENTIRELY SURE I CAN ANSWER YOUR QUESTION, BUT I DO THINK THAT A LOT OF THE IT COMES FROM THE OUTPUT PATHWAY SPECIFICITY. >> SO THE CONNECTIVITY STUDIES ARE IMPRESSIVE, THE STUDIES YOU CAN DO WITH CRIMSON, BUT I'M WONDERING IS IT SPOBL TO DO SOME SORT OF FUNCTIONAL OCCLUSION TEST, FOR EXAMPLE, YOU SHOWED EXPERIENCE WHERE YOU SILENCE THE INPUTS TO THE MUSHROOM BODIES, THE DOPA MINERGIC NEURONS AND THEN YOU SHOW OTHER EXPERIMENTS WHERE YOU SHOW OTHERS CAN YOU SUM ULTIMATELY TANIOUSLY SHUT 1 DOWN WHILE YOU REACT-OR ACTIVATE THE OTHER AND SEE NO CHANGE IN THE BEHAVIOR? YOU WOULD SEE WITH JUST SHUTTING DOWN 1? DOES THAT MAKE SENSE? NOT REALLY? >> I THINK IN PRINCIPLE YOU CAN BUT 1 OF THE THINGS THAT'S REALLY CLEAR FROM THE WORKER SYSTEM THE SPECIFICITY OF THE ORDERS, THEY'RE ACTUALLY DIRECT, THE PROCESS THAT IS GENERATED SO, IF I ACTIVATE FOR EXAMPLE, GENTLEMENNA 2 ALPHA PRIME 1 MUSHROOM BODY OUTPUT NEURON, IT WOULDN'T KNOW THE DIFFERENCE BETWEEN THE CS- AND THE CS+, BUT THE CANYON CELL DRIVE TO THE OVERALL ENSEMBLE IS CHANGED VIA LEARNING AND FROM THERE YOU GET A LAB MUSHROOM BODY OUTPUT, SO GOING BACK TO THE QUESTION, RIGHT? SO I DON'T THINK IT WOULD BE AN INFORMATIVE EXPERIMENT TO DO THAT. I THINK THE SPECIFICITY JUST SIMPLY WOULDN'T BE THERE. >> SCOTT, SO, VERY NICE TALK AND I'M--THE MECHANISMS, YOU DESCRIBED IN SO FAR AS I UNDERSTAND THEM, VERY ELEGANT, BUT-- >> PLEASE DON'T BELIEVE IT. >> [LAUGHTER] >> BUT I AM SURPRISED A BIT AND I'M WONDER HOW YOU THINK ABOUT 1 THING IN PARTICULAR. SO IT SEEMS LIKE, THE CS+ AND MINUS SORT OF HAVING EQUAL DEMANDS ON THE FLIES ATTENTION AND COGNITIVE ABILITIES AND THE FLY IS ACTIVELY TRYING TO LEARN WHAT IS REWARDING AS WELL AS WHAT IS NOT REWARDING. AND THAT KIND OF MAKES SENSE BUT ON THE OTHER HAND MOST CHOICES IN LIFE MOST EXPERIENCES IN LIFE ARE NOT BINARY WHERE YOU'RE SOWTER OF CHOOSING BETWEEN 2 THINGS, 1 OF WHICH IS IRRELEVANT AND NOT REWARDED AND 1 WHICH IS REWARDED AND SEEMS TO ME THE PREPONDERANCE OF THINGS THAT ONCE EXPERIENCES ARE NOT REWARDING, AND ONLY A FEW CERTAIN THINGS TEND TO BE REWARDING AND IT SEEMS TO ME IT WOULD LEAD TO AN EXPLOSION THAT YOU WERE TRYING TO KEEP TRACK OF EVERYTHING THAT WASN'T IN A REWARDING AND AND THOSE FEW THINGS, SO HOW DO YOU THINK ABOUT THAT? >> IT'S A GREAT QUESTION. THEN, SEE 1 OF THENINGS I HAVEN'T ALLUDED TO AND I'M GRATEFUL FOR YOU ASKING THIS QUESTION, IS THAT, THERE ARE MANY THINGS THAT THAT DRIVE MEMORY CONSOLIDATION, RIGHT? SUCH THAT IT'S STUDIED BY MANY PEOPLE AND 1 OF THE THINGS THAT'S A CURRENT TOPIC OF INTEREST IS SOMETHING THAT PEOPLE CALL BOUNDARY CONDITIONS, THESE ARE INSTANCES THAT DRIVE RECONSOLIDATION AND THOSE THAT DON'T, RIGHT? SO WHETHER IT'S REALLY SPECIFICALLY THE CS- THAT DRIVES IT OR NOT IS 1 OF THOSE POTENTIAL BOUNDARY CONDITIONS, THINGS THAT DO AND THE THINGS THAT DON'T SO WE LOOKED AT THIS AND I TOLD YOU THAT WITH EXTENSION, THE CS+ DRIVES EXTENSION, THE CS- DOESN'T AND THE [INDISCERNIBLE] DOESN'T. YOU COULD SAY, OKAY, AS YOU DID, IT'S COSTLY TO REMEMBER EVERYTHING THAT YOU DIDN'T REWARD, RIGHT? BUT DURING TRAINING THE FLIES GET THE CS, THEY'RE ALSO IN THE TRAINING MACHINE AND THERE'S AIR PUFF, THERE'S MANY CONCEPTS YOU MIGHT--OKAY, WELL, WHY DON'T THEY REMEMBER THESE, THESE DRIVE RECONSOLIDATION, SO THE CS MINUS DRIVES RECONSOLIDATION AND AIR STREAM DRIVES RECONSOLIDATION AND MAYBE LOADING THEM IN THE MACHINE DRIVE RESCONSOLIDATION BUT THE KEY BOUNDARY THAT DOESN'T IS A NOVEL ORDER. IF YOU GIVE THEM A NOVEL ORDER IT DOESN'T TRIGGER RECONSOLIDATION, SO THERE'S SOMETHING SPECIFIC ABOUT WHAT THEY REMEMBER FROM THE TRAINING EXPERIENCE BUT IT'S NOT--THERE ARE THINGS THAT THEY CONSIDER NOVEL THAT DON'T TRIGGER THESE THINGS NOW CAN I SAY, IS THAT ADAPTIVE BECAUSE IT'S INCREDIBLY COSTLY TO DO THAT, I CAN'T--I SIMPLY CAN'T ANSWER THAT QUESTION, RIGHT? WOULD WE SEE THIS IN A NONBINARY PAIR IN TRAINING EXPERIMENT? WE DON'T KNOW, WE ONLY SAW THIS PHENOMENON OF RECONSOLIDATION BECAUSE IT'S TRIGGERED BY THE CSMINUS AND WE LOOK AT THESE BOUNDARIES BUT I CAN'T ANSWER THE QUESTION, IT'S A GOOD 1 BUT WE JUST--WE'RE UNABLE TO ANSWER IT AT THE MOMENT. >> IF THERE ARE NO OTHER QUESTIONS, THANK YOU. [APPLAUSE ]