GOOD AFTERNOON. YOU ALL KNOW WHY WE ARE HERE TODAY. THE REASON WHY SLEEP WAS SUGGESTED BY MANY PEOPLE AND ENTHUSIASTICALLY SUPPORTED BY MYSELF IS SLEEP IS ONE OF THOSE THINGS WE TAKE FOR GRANTED UNLESS WE HAPPEN TO HAVE SOME DISORDER AFFECTING. BUT YOU KNOW, IT'S LIKE BREATHING AND ALL KINDS OF NORMAL ACTIVITIES. BUT IT'S A GREAT MYSTERY AND AN ENORMOUS CHALLENGE. IN THIS MODERN ERA, SOME WINDOWS ARE OPENING AND DOORS ARE OPENING AND SOME CLARITY IS COMING TOWARD HELPING TO ANSWER SOME OF THE AGE-OLD QUESTIONS. I PUT ON THE WEBSITE SOME MATERIAL BUT I ALSO PUT A REFERENCE IN THIS NOW NORMALLY WE DON'T OFFERED BOOKS AS PART OF THE COURT. BUT HIS WORK ON MEMORY, THIS IS AN AMAZING BOOK WHICH TOUCHES VERY MUCH ON CONSCIOUSNESS. IN FACT, CONSCIOUSNESS FROM THE PERSPECTIVE OF ART AND VIENNA AT THE TURN OF THE LAST CENTURY, MUSIC, THE BIRTH OF COGNITIVE PSYCHOLOGY, THE CHANGES IN MEDICINE AND REALLY SOME POPULAR SCIENCE LEVEL HARD CORE NEUROBIOLOGY. SO WE LOOK FOR LIVER DOCTORS LIKE ME. THIS WAS A WONDERFUL THING TO READ. I PUT THE REFERENCE ON THE WEBSITE IN CASE ANY OF YOU ARE INTERESTED. IT WILL KEEP YOU BUSY FOR SEVERAL MONTHS. IT'S TAKEN ME OVER TWO AND I HAVEN'T FINISHED IT YET. BUT THAT'S A REFLECTION MORE OF MY LIMITED UPBRINGING BUT IT'S REALLY WRITTEN ELEGANTLY. SO WHY, WHAT, HOW AND WHEN. TODAY'S TWO SPEAKERS WE'RE FORTUNATE ONCE AGAIN HAVING HERE AT NIH, ALWAYS A RAID OF PEOPLE IN ALMOST ANY AREA OF MAJOR INTEREST THAT YOU CAN THINK OF. TODAY'S SPEAKERS, FIRST WILL BE CAROLYN BEEBE SMITH, WHO TOOK HER DEGREE AT THE UNIVERSITY OF LONDON. AND CAME HERE WHEN? CAME TO NIH IN 2000. 2011. THAT'S RIGHT YOU WORK AT THE MINH, RIGHTED. AND SHE IS A SENIOR INVESTIGATOR IN THE LABORATORY OF CEREBRAL METABOLISM. AND AS YOU WILL SEE, SHE IS EXPLOITING NEW TECHNOLOGIES TO STUDY SLEEP AND OTHER DISORDERS IN EXPERIMENTAL TRANSGENIC ANIMALS AND THEIR BRAINS. AND OUR SECOND SPEAKER IS SUSAN HARBISON WHO WHEN I ASKED HER WHY DID SHE HAD AN ORIGINAL DEGREE IN AERONAUTICAL ENGINEER AND THEN MADE A DRAMATIC CHANGE AND TOOK A PH.D. AND EMERGED WITH EXPERTISE IN DROSOPHILA, PARTICULARLY TAKING ADVANTAGE OF THE UNIQUE GENETICS OF DROSOPHILA TO STUDY PROBLEMS RELATED TO SLEEP AND OTHER DISORDERS. SUSAN JOINED THE HEART INSTITUTE HERE IN 2012 AS A NEUROSTAFF AND TENURE TRACK INVESTIGATOR. FOR THOSE OF YOU WHO DON'T KNOW, THIS IS AN EXTRAORDINARY PROGRAM THAT THE NIH HAS WHICH IS HIGHLY COMPLETIVE AND IT'S REALLY THE CREME DE LACREME. SO TIME REALLY STARTS I THINK IN POETRY AND LITERATURE WHERE DEATH AS A MANIFESTATION OF SLEEP ATTRACTED THE ATTENTION IN THE TEMPEST THAT AS A FORM OF SLEEP AND SLEEP AS A FORM OF DEATH, THOSE THINGS WERE INTIMATELY LINKED. IT'S A VERY INTERESTING QU. THEY PROBABLY BELIEVED THAT WHEN YOU WENT TO SLEEP, YOUR BRAIN WENT DEAD. VERY IMPORTANT CONCEPT. AND THEN COMES THE AREA OF THE DEVELOPMENT OF ANESTHESIA, CHLOROFORM, ETHER. AND PEOPLE ALSO WENT UNCONSCIOUS OR THEY MAINTAINED THEIR VITAL SIGNS IF THEY DIDN'T INHALE TOO MUCH. AND WHAT WAS THAT ANESTHESIA, WAS THAT THE SAME AS SLEEP? IS IT THE SAME AS SLEEP. THERE'S A HUGE LITERATURE THAT INITIALLY SAID IT WAS SLEEP, INDUCED SLEEP. AND THEN THERE WERE OTHERS WHO CAME ALONG AND SAID NO NO NO, THIS IS DIFFERENT. AND THEN WE GET INTO NOW THE LATE 19TH CENTURY WHERE CONCEPTS OF CONSCIOUSNESS, SLEEP AND THEN LATER THE INTRODUCTION OF ELECTROENCEPHALOGRAPHY AS REALLY THE ONLY TOOL ASIDE FROM PATHOLOGY. BUT IN PATHOLOGY SHE'S DEAD SO YOU DON'T KNOW WHAT'S GOING ON, YOU MAY HAVE KNOWN WHAT HAPPENED. SO IT WAS REALLY I THINK THE ELECTROENCEPHALOGRAM THAT SHOWED THE VARIOUS WAVE PATTERNS WITH PEOPLE FALLING ASLEEP. NON-EYE MOVEMENT SLEEP AND WAKING PEOPLE UP AND DISCOVERING WHEN THEY WERE DREAMING. AND THIS SORT OF SET THE BASIS OF ANOTHER IMPORTANT CONCEPT AND THAT IS THE BRAIN IS NOT DEAD WHEN YOU'RE ASLEEP. IT'S VERY MUCH ALIVE. BUT THAT MAY SOUND SIMPLE, BUT HOW IT'S ALIVE IS STILL THE MYSTERY TODAY. AND THEN BEGINNING IN THE EARLY 20TH CENTURY, AND ACCELERATING LIKE CRAZY IN THE MODERN WORLD IS THE DEVELOPMENT OF COGNITIVE PSYCHOLOGY. THAT'S WHERE THE BOOK THE AGE OF INSIGHT IS A BRILLIANT DISCUSSION OF THE ROLE OF THE COGNITIVE PSYCHOLOGISTS INCLUDING FREUD AND HIS COLLEAGUES AND ARTISTS AND POST AND WRITERS IN EARLY NEUROBIOLOGISTS. THE RECOGNITION OF EMOTIONS AND NSCIOUSNESS IN THEIR RELATIONSHIP AND THEN OF COURSE WE ZOOM UP TO THE PRESENT TIME WHEN SAILOR ANATOMY IS NOW ACCESSIBLE IN LIVING TISSUES, MORE SO AND MORE SO THROUGH THE USE OF ANATOMIC FUNCTIONAL METABOLIC SCANS. LITERAL DISSECTION. AND THEN THAT BECOMES COUPLED WITH YOU MOVE TO ANOTHER STATION DROSOPHILA AT THIS CASE WHERE THE GENETICS ARE ALL THERE. HOW DO YOU USE THAT TO EXPLAIN PHENOMENON WITH SLEEP. THE SAME THING IS GOING TO APPEAR IN A WALS LECTURE LATER ON NEXT MONTH WHEN EN RIQUE COMES AND TALKS TALKING ABOUT ALCOHOL CALL COMES, ADDICTION AND SO FORTH. SO ENOUGH ARE YOU SPEAKING FIRST? >> OKAY, I'LL PRESENT. MY VOICE IS A LITTLE ROUGH. SO I JUST WANT TO THANK YOU FOR THE INVITATION, THIS IS FUN TO GET TOGETHER TO TALK ABOUT SLEEP AND THANK YOU ALL FOR COMING. SO I AM GOING TO TELL YOU A LITTLE BIT ABOUT SLEEP IN GENERAL, SOME OF THE BIG IDEAS ABOUT SLEEP AND THEN EVENTUALLY GO INTO THE STUD[INDISCERNIBLE]. SO WE TALKED JUST NOW A LITTLE BIT ABOUT [INDISCERNIBLE]. SO WHEN YOU'RE ASLEEP, THERE'S THE CHARACTERISTICS OF THE [INDISCERNIBLE] AND A REDUCED RESPONSE TO SENSORY [INDISCERNIBLE] NOW WHEN PEOPLE ARE INTERESTED IN SLEEP AND INTERESTED IN A LOT OF ASPECTS, ONE THERE'S MANY [INDISCERNIBLE]. IN ADDITION IT'S IN OUR CULTURE WE'RE VERY INTERESTED [INDISCERNIBLE] WHAT HAPPENED THEN YOU DON'T GET ENOUGH SLEEP. AND THEN IT'S KIND OF THE ACADEMIC QUESTION [INDISCERNIBLE] WHAT'S HAPPENING TO US [INDISCERNIBLE] AND THIS OF COURSE A BIG, THIS IS GRAND RESEARCH QUESTION. SOME OF THE DISORDERS OF SLEEP. THE ABILITY TO GO TO SLEEP OR STAY ASLEEP OR SLEEPING TOO MUCH ARE [INDISCERNIBLE] SOMETHING ELSE WRONG WITH YOUR BRAIN [INDISCERNIBLE] BUT PEOPLE WHO ARE DEPRESSED CAN HAVE EITHER OF THESE [INDISCERNIBLE]. SO SOME OF THE DISORDERS [INDISCERNIBLE] THEN WE CAN ASK [INDISCERNIBLE] AND THEM YOU CAN'T SLEEP [INDISCERNIBLE] MORE DEPRESSED. ANOTHER ONE THAT I'M SURE YOU'VE HEARD ABOUT IS RESTLESS LEG SYNDROME OCCURS MORE IN OLDER PEOPLE. THEY HAVE SENSATIONS [INDISCERNIBLE] HAVE TO MOVE AND THEN THEY WAKE UP. ANOTHER ONE IS BRUXISM, WHICH IS CRUNCHING YOUR TEETH. WHEN THIS IS VERY EXTREME, IT CAN WAKE YOU UP AND OF COURSE IT CAN ALSO GO DOWN INTO YOUR TEETH SO YOUR DENTIST WILL BE TALKING TO YOU ABOUT THAT IF YOU HAVE THAT PROBLEM. THE NEXT ONE, SLEEP APNEA IS ANOTHER ONE THAT'S BEEN A LOT IN THE NEWS IT. OCCURS MORE IN MIDDLE-AGE THE MEN, PARTICULARLY OVERWEIGHT MIDDLE-AGED MEN. WHAT HAPPENS HERE IS THERE IS ACTUALLY A SLAPS OF THE AIRWAYS. LOUD SNORING WHICH IS ONE OF THE THINGS YOU CAN DETECT WITH SOMEBODY WHO HAS SLEEP APNEA. AND THEY ACTUALLY, THEIR BLOOD OXYGEN LEVEL WILL GO DOWN AND THAT WAKES THEM UP. SO THESE PEOPLE CAN WAKE UP A HUNDRED TIMES A NIGHT. SO YOU CAN SEE HOW THAT INTERFERES WITH THEIR ABILITY TO SLEEP. AND OF COURSE NARCOLEPSY IS ANOTHER ONE I'M SURE YOU HEARD OF. THERE'S SOMEBODY IN RESPONSE TO EMOTION THEN ALL OF A SUDDEN LOSES ALL MUSCLE TONE, COLLAPSES AND GOES TO SLEEP. A PARTICULARLY TERRIBLE AFFLICTION. EVEN WORSE AFFLICTION IS ONE THAT AGAIN I'M SURE YOU'VE ALL HEARD OF IS FATAL FAMILIAL INSOMNIA. THIS IS A DISEASE THAT IS AN AUTOSOMAL DOMINANT INHERITANCE WHERE THERE'S A MUTATION ON THE GENE FOR PRION IN CHROMOSOME 20. THIS IS FIRST DESCRIBED IN 1986, AN ITALIAN FAMILY [INDISCERNIBLE] IT'S A PROGRESSIVE UNTREATABLE INSOMNIA. PEOPLE THAT ARE AFFLICTED WITH THIS DISEASE AND DEVELOPED IN INSOMNIA, THEY DIE WITHIN FAIRLY SHORT TIME. AND THIS CAN STRIKE WHEN SOMEBODY IS IN THEIR 20'S OR WHEN THEY MIGHT BE FINE UNTIL THEY'RE IN THEIR 60'S. SO WHAT WE SEE IN THEIR BRAINS IS THAT THERE'S A CHARACTERISTIC DEGENERATION PARTICULARLY IN THE THALAMUS AND ALSO THIS SPONGY FORM ENCEPHALOPATHY WHICH YOU CAN SEE HEAR IN THE THALAMUS AND THEN THE CORTEX. HERE'S THE CORTEX AND THE THALAMUS. THAT'S ACCOMPANIED BY THE GLIOSIS. THIS IS A DISEASE THAT'S SO RARE. NOW, WE ALL KNOW FROM PERSONAL EXPERIENCE AGAIN THAT THERE ARE THE EFFECTS OF SLEEP DEPRIVATION, THERE ARE SOME INTERESTING STUDIES, IN RATS, FOR INSTANCE -- HAS SHOWN IN RATS THAT ARE TOTALLY DEPRIVED OF SLEEP, THAT THEY WILL DIE IN ABOUT THREE WEEKS. SO IT'S AGAIN, FATAL, NOT SLEEPING. AND THERE WAS A STORY FROM MY, THAT I REMEMBER FROM MY YOUTH MAYBE SOME PEOPLE IN THE AUDIENCE MAY REMEMBER OF A DJ IN NEW YORK CITY NAMED PETER TRIP WHO HE WAS VERY POPULAR FOR, WITH EVERYBODY. HE WAS THE GUY WHO INVENTED THE TOP 40. AND HE RAISED MONEY FOR MARCH OF DIMES THAT HE WOULD STAY AWAY FOR OVER 200 HOURS. AND HE ACTUALLY HAD A LITTLE GLASS BOOTH IN TIMES SQUARE AND IT WAS A BIG PRODUCTION. THERE WAS A LOT OF PUBLICITY ABOUT IT. AND HE STARTED TO SHOW SIGNS OF PSYCHOTIC BEHAVIOR AFTER A FEW DAYS OF THAT. HE DID DO IT. HE STAYED AWAKE FOR 200 DAYS. HE HAD TO HAVE SOME MEDICATION TO KEEP HIM AWAKE, I PRESUME. I DON'T KNOW WHAT THEY GAVE HIM BUT ANYWAY, THE INTERESTING THING AFTERWARDS IS THAT HE WAS NEVER THE SAME. >> [INDISCERNIBLE] >> RIGHT. HE ACTUALLY LOST HIS JOB, HE GOT INVOLVED IN CRIME. HIS LIFE KIND OF FELL APART. IT WASN'T THAT IT HAD TO DO WITH THE SLEEP DEPRIVATION BUT THAT WAS A DRAMATIC EXAMPLE IN THE NEWS OF SOMEBODY WHO HAD STARVED HIMSELF OF SLEEP. NOW FOR MOST OF US, WE GO AROUND I THINK WITH OUR SLEEP TANK ON ABOUT A QUARTER, MAYBE A HALF, SOME OF THE GOOD ONES OF US. WE'RE USUALLY NOT VERY GOOD ABOUT IN OUR CULTURE ABOUT SLEEP HYGIENE. WE KIND OF UNDER VALUE SLEEP. AND THERE ARE SOME PRETTY IMPORTANT CONSEQUENCES OF THAT. THERE ARE PSYCHOLOGICAL ONES WHICH I THINK YOU PROBABLY KNOW ABOUT, YOU REDUCE YOUR PERFORMANCE, YOU HAVE SLOWER REACTION TIME. YOU ALSO INCREASE YOUR RISK FOR DEPRESSION AND ANXIETY. AND YOU ALSO SUPPRESS YOUR IMMUNE SYSTEM. AND THEN YOU INCREASE YOUR RISK FOR A NUMBER OF DISEASES. AND EVEN FOR OBESITY. SO SLEEP HAS AN IMPORTANT FUNCTION. IF IT'S JUST BEING DEPRIVED OF SLEEP IS GOING TO PRODUCE SUCH EFFECTS. BUT SLEEP AS HAS BEEN SAID IN THE INTRODUCTION IS KIND OF A SINGLE STATE. THERE ARE NUMEROUS STAGES OF SLEEP. AND I AM THOUGHT GOING TO MAKE THIS INTO A LESSON IN POLY SOMNOGRAPHY. THE STAGES CAN BE DEFINED BY POLY SOMNOGRAPHY WHERE THE ELECT GRAM IS PUT AROUND YOUR SCALP. SO NOW IT'S UNDER YOUR EYES TO MONITOR YOUR EYE MOVEMENT AND ELECTROMYOGRAM TO LOOK AT YOUR MUSCLE TENSION. IT'S USUALLY UNDER THE CHIN. WHEN YOU'RE AWAY YOU HAVE THESE CHARACTERISTIC HIGH FREQUENTLY WAVES AND THE EEG. LOTS OF EYE MOVEMENTS AND LOTS OF MUSCLE TENSION. WHEN YOU FIRST FALL ASLEEP YOU GO INTO STAGE ONE WHICH IS KIND OF A TRANSITIONAL STAGE. AND THE EEG BECOMES A LITTLE LOWER FREQUENCY. EYE MOVEMENTS ARE SLOW AND THE MUSCLE TENSION IS MEDIUM. IN STAGE TWO, YOU'RE GETTING A LITTLE DEEPER. NOW THE EEG HAS SOME INTERESTING WAVE FORMS, THESE COMPLEXES AND SLEEP SPINDLES WHICH JUST INDICATE THAT YOU'RE GETTING INTO DEEPER SLEEP. EYE MOVEMENTS HAVE CEASED AND AGAIN THE EMG IS KIND OF MEDIUM TENSION. WHEN YOU GO INTO STAGES THREE AND FOUR, THAT'S NOW YOU'RE INTO A DEEP SLEEP, A SLOW WAVE SLEEP. AND HERE, THE EEG HAS THESE CHARACTERISTIC WAVES THAT ARE OF LOW FREQUENCY AND HIGH AMPLITUDE AND MORE IN STAGE FOUR THAN STAGE THREE BUT THEY'RE VERY SIMILAR IN THE TWO. YOU SLEEP MORE. AGAIN, EYE MOVEMENTS HAS CEASED AND THE EMG IS LOW. AND FINALLY, REM WHICH WAS THE LAST STAGE TO BE DISCOVERED. AND THERE, THE EEG LOOKS A LITTLE MORE LIKE LIGHT SLEEP OR AWAKE, AND YOU SEE THESE RAPID EYE MOVEMENTS. AND ONE OF THE MOTION CHARACTERISTIC THINGS IS THE MUSCLE TENSION IS JUST ABOUT GONE. SO THEY'RE VERY VERY RELAXED. SO JUST TO KIND OF SUMMARIZE THE, WHAT HAPPENED WITH SLEEP OVER THESE STAGES. WHEN YOU'RE AWAKE YOU HAVE A DESYNCHRONIZED FIRING OF NEURONS. WHEN YOU GO INTO NON-REM SLEEP, THE FIRING OF THE NEURONS IS MORE SYNCHRONIZED. SO YOU'VE GONE FROM THE STATE WHERE NEURONS ARE ALL KIND OF DOING THEIR OWN THING AND THEIR CONSCIOUS BRAIN TO A STATE WHERE THEY'RE MUCH MORE FIRING TOGETHER AND A LITTLE MORE OBLIVIOUS TO WHAT'S GOING ON OUTSIDE. AND IN REM SLEEP, THEY'RE DESYNCHRONIZED AGAIN. SO THE STATE IS QUITE DIFFERENT FROM THE NON-REM SLEEP. OVER A NIGHT, YOU GO IN STEPS. THIS IS HOURS OF SLEEP. SO YOU GO IN STEPS TO THIS DEEP STAGE FOUR SLEEP. AND THEN HAVE A FEW CLIMBS BACK UP TO REM. SO IN THE BEGINNING OF THE NIGHT, YOU SPEND A BIGGER PROPORTION OF TIME IN DEEP SLEEP AND SLOW SLEEP, STAGE THREE AND FOUR. AND SOME SMALLER TIME IN REM. AND THEN AS THE NIGHT PROGRESSES, YOU START TO SPEND MORE TIME IN STAGE TWO AND REM SLEEP. SO THIS IS KIND OVER THE PROGRESSION OF THESE WAVES OF SLEEP OVER THE NIGHTTIME. NOW OVER YOUR LIFETIME, SLEEP ALSO CHANGES. WE KNOW THAT BABIES SPEND A LOT OF TIME WITH SLEEP. MAYBE 16 HOURS A DAY OF SLEEP. AND ABOUT HALF OF THAT IS IN REM AND HALF IN NON-REM. PRENATALLY THEY SPEND EVEN MORE TIME IN REM. AND THEN AS THEY DEVELOP DURING EARLY CHILDHOOD, THE REM DECREASES BUT ACTUALLY THE NON-REM COMPONENT GOES UP. AND IN THE FIRST FEW YEARS OF LIFE IT STAYS PRETTY CONSTANT WHERE THE REM IS DECREASING. AS WE GET OLDER, WE SPEND LESS AND LESS TIME WITH SLEEP AND LESS AND LESS TIME IN REM. SO THERE ARE SEVERAL PROCESSES THAT DRIVE SLEEP. ONE IS CIRCADIAN. THE CIRCADIAN 24-HOUR CLOCK AND THE OTHER IS A HOMEOSTATIC DRIVE. LIKE ANY APPETITE, IT'S JUST LIKE BEING HUNGRY WHEN WE'RE AWAKE FOR A LONG TIME WE GET SLEEPIER AND SLEEPIER THE LONGER YOU ARE AWAKE. SO THIS JUST ILLUSTRATES THAT OVER THE AWAKE AND ASLEEP PERIOD THAT YOU HAVE THIS CIRCADIAN PATTERN. BUT THE HOMEOSTATIC DRIVE, THE LONGER YOU'RE AWAKE AND THE BIGGER DRIVE TO SLEEP. AS YOU'RE SLEEP THAT DIMINISHES WHEN YOU WAKE UP. SO THE CIRCADIAN DRIVE IS THROUGH A PART OF THE ANTERIOR HYPOTHALAMUS. THERE ARE THESE LITTLE NUCLEI IN THE ANTERIOR THALAMUS CALLED -- THERE'S A SIGNALS FROM THE RETINA TO THE SCM. SO LIGHT WILL STIMULATE THE RETINA AND THEN STIMULATE THE SCM. AND THAT WILL STIMULATE THE AREA OF THE HYPOTHALAMUS THAT THEN ACT TO ACTIVATE BRAIN STEM ACTIVATING SYSTEMS. AND THIS JUST SHOWS THE CIRCADIAN PATTERN OF AN ANIMAL THAT IS RUNNING WITH HIM AND HE'S BEEN TRAINED TO LIGHT. HE IN FACT WAS QUIET IN THE LIGHT AND NOT IN THE DARK. THE HOMEOSTATIC DRIVE, ONE OF THE MAJOR PLAYERS IS ADENOSINE AND THIS IS A PRODUCT OF CELLULAR ENERGY METABOLISM. AND IT ACCUMULATES DURING WAKEFULNESS. IT ALSO ACTS AS A NEUROTRANSMITTER. SO YOU CAN SEE HOW ADENOSINE CAN ACT ON THAT. HOW THE DRIVE TO SLEEP WITH ADENOSINE GOING ON. THIS IS JUST TO SHOW YOU THAT THERE ARE THESE ACTIVATING SYSTEMS IN THE BRAIN THAT ARE FEEDING UP FROM THE BRAILLE S TEM -- BRAIN STEM TO THE POND TO ACTIVATE THE CORTEX, THALAMUS AND SORT OF KEEP YOU ALERT, AROUSED AND THERE ARE THE THREE MAJOR SYSTEMS RETICULAR ACTIVATING SYSTEM WHICH IS THE CHOLINERGIC SYSTEM -- WHICH IS ANOTHER PROJECTION. SO WE SAID AT THE OUTSET THAT ONE OF THE BIG QUESTIONS IS WHY DO WE NEED TO SLEEP. IT'S CLEAR WE NEED TO SLEEP, BUT WHY. SO ONE OF THE IDEAS IS THAT DURING WAKEFULNESS, THERE ARE, THERE'S AN ACCUMULATION OF METABOLIC PRODUCTS THAT HAVE TO BE CLANKED UP, TAKEN CARE OF. AND THERE'S ALSO MAINTENANCE THAT NEEDS TO BE DONE ON THE NERVOUS SYSTEM JUST TO TAKE CARE OF WEAR AND TEAR JUST AS YOU WOULD A ROAD SYSTEM. AND THIS, MAYBE THIS IS BEST DONE WHEN THE BRAIN ISN'T DOING OTHER THINGS. SO THAT'S CALLED THE RESTORATION HYPOTHESES, VERY SIMPLE IDEA, VERY DIFFICULT TO PROVE. ANOTHER IDEA THAT HAS BEEN DISCUSSED A LOT IN THE LAST 10-15 YEARS IS THE INFLUENCE OF SLEEP OR SYNAPTIC REMODELING. AND SO THIS PERTAINS MAINLY AGAIN TO THE NERVOUS SYSTEM AND IN THE NERVOUS SYSTEM ONE OF THE KEY ATTRIBUTES OF THE NERVOUS SYSTEM IS THIS PLASTICITY. THE PLASTICITY IS THE ABILITY OF A NERVOUS SYSTEM TO ADAPT TO CHANGES IN ENVIRONMENTAL INPUT. AND IN DEVELOPMENT PLASTICITY IS OF COURSE VERY IMPORTANT FORGETTING PATHWAYS ORGANIZED. AND BUT IT DOESN'T ONLY OCCUR IN DEVELOPMENT, IT ALSO OCCURS IN ALL OF US OR WE WOULDN'T LEARN AND REMEMBER ANYTHING. SO PLASTICITY IS VERY IMPORTANT PROCESS. AND WHAT THEY'RE LEARNING IS THAT THERE IS AN INFLUENCE OF SLEEP ON PLASTICITY. SO I'M GOING TO FOCUS ALL THE REST OF MY REMARKS ON THIS ISSUE. NOW TO TALK ABOUT DEVELOPMENTAL PLASTICITY, ONE OF THE KEY SYSTEMS THAT'S STUDIED BECAUSE IT'S A VERY ROBUST SYSTEM IS PLASTICITY OF THE BINOCULAR VISUAL SYSTEM. SO ANIMALS WITH BINOCULAR VISION LIKE A CAT, A MONKEY, WE HAVE INPUT FROM BOTH EYES AND THAT INPUT, IT ORIGINATES SAYING THE LEFT PART OF THE VISUAL FIELD WILL GO THROUGH BOTH EYES. THE INPUT GOES THROUGH THE THALAMUS AND THEN THE PRIMARY VISUAL CORTEX. THE INPUT FROM THE TWO EYES REMAIN SEGREGATED ALL ALONG THIS PATHWAY SO THROUGH THE THALAMUS AND IN THE CORTEX IN THE FORM OF THESE OCULAR DOMINANTS. SO FROM THE LESS VISUAL FIELDS IT WILL ACTIVATE THE RIGHT PRIMARY VISUAL CORTEX AND THE LEFT VISUAL FIELD, RIGHT VISUAL FIELD SORRY WILL ACTIVATE LESS. SO WHAT WE'VE LEARN BOARD OF DIRECTOR THIS SYSTEM AND THIS IS LARGELY FROM THE WORK OF -- WHICH THEY WITHIN THE NOBEL PRIZE IS THAT DURING DEVELOPMENT, IF YOU BLOCK THE INPUT FROM ONE EYE DURING THE TIME THAT THIS IS DEVELOPING YOU WILL ACTUALLY END UP WITH A CORTEX THAT IS RESPONSIVE NOW ONLY TO THE OPEN EYE. AND PEOPLE HAVE SHOWN THIS IN CATS AND MONKEYS. SO WE IN NEUROSCIENCE USE THIS SYSTEM A LOT TO STUDY PLASTICITY. AND MARKUS FRANK LAB LOOKED AT THIS PROCESS IN CATS AND KITENS, AND HE ASKED THE QUESTION DOES SLEEP HAVE ANY EFFECT ON THIS PLASTIC RESPONSE. SO IN ORDER, I'LL JUST TELL YOU JUST FOR A SECOND WHAT THESE ARE SO THAT IT DOESN'T BECOME OVERWHELMING. BUT THESE ARE ESSENTIALLY THEIR PERCENT RECORDING OF NEURONS THAT ARE RESPONDING TO LIGHT FLASH FROM GOING THROUGH ONE EYE OR THE OTHER. AND HE'S INDICATED HERE THIS IS THE OPEN EYE, THIS IS THE DEPRIVED EYE. SO HE TOOK A GROUP OF CATS AND THEY HAVE BINOCULAR VISION TO START WITH AND THEN THEY HAD SIX HOURS OF MINOCULAR IN THE VISION OF THE LIGHT. AND THEY TOOK A HISTOGRAM OF THIS DOMINANCE. YOU CAN SEE THERE'S A LITTLE TENDENCY FOR DOMINANCE TO BE HIGHER OUTPUT AND LOWER IN THE DEPRIVED EYE. HE TOOK ANOTHER GROUP, DID THE SAME THING BUT THEN AT THE END OF THIS SIX HOURS OF WAKEFULNESS IN THE LIGHT HE PUT THEM IN THE DARK AND LET THEM GO TO SLEEP. AFTER SIX HOURS DID ANOTHER RECORDING SESSION OR DID A RECORDING SESSION. NOW HE'S SHOWN THAT'S PRETTY DRAMATIC THAT MORE CELLS ARE RESPONDING TO THE OPEN EYE AND FEWER TO THE CLOSED EYE. THE NEXT QUESTION IS WELL IS THAT JUST BECAUSE THEY HAD ANOTHER SIX HOURS. AND SO HE TOOK ANOTHER GROUP AND STUDIED THEM WHILE THEY REMAINED AWAKE SO THEY KEPT HIM AWAKE BUT IN THE DARK. AND HERE YOU CAN SEE HE DIDN'T GET THIS EFFECT. SO THIS IS JUST TIME. IT SEEMS TO HAVE SOMETHING TO DO WITH SLEEP. AND HE TOOK A FULL SCOOP AND KEPT THEM AWAKE BUT IN THE LIGHT. SO NOW THEY'VE HAD 12 HOURS OF MIN OCULAR. SO THE SLEEP ENHANCED THE PRECEDING PERIOD OF MINOCULAR DEPRIVATION AND HINTS THAT RESPONSE IN THE VISUAL CORTEX. SO ANOTHER QUESTION IN HIS LAB IS WHAT PROCESS DOES THAT DEPEND ON. MOST OF THE THINGS THAT WE ARE INTERESTED IN PLASTICITY IS BROKEN SENSE -- BECAUSE WE KNOW PLASTICITY IS USUALLY BLOCKED BY PROTEIN SYNTHESIS INHIBITORS. HE USED, HE STUDIES THE SAME SYSTEM BUT HE USED AN INHIBITOR CALLED RAPAMYCIN WHICH BLOCKS THE ENTORE PATHWAY THE REGULATOR OF PROTEIN SYNTHESIS. AND SO IN THIS SERIES OF HISTOGRAMS, FIRST YOU CAN SEE WHAT IT LOOKS LIKE WHEN A KITTEN HAS STUDIED THAT HAS BINOCULAR VISION. THERE WAS A GROUP THAT GOT VEHICLE AND ANOTHER GROUP HERE TWO GROUPS GOT RAPAMYCIN THAT'S APPLIED TO THE CORTEX. WHEN HE DID MINOCULAR DEPRIVATION WHETHER THEY GOT RAPAMYCIN OR NOT, HE REALLY DIDN'T GET MUCH OF A CHANGED IN HISTOGRAM. THEY SLEPT AT THE TIME THEY GOT THE VEHICLE RAN MYOSIN. IN THE TREATED ANIMALS HE GOT THAT SAME SHIFT THAT HE SAW IN THE PREVIOUS EXPERIMENT. BUT WHEN HE DID THE SAME THING BUT TREATED THEM WITH RAPAMYCIN, IT WAS BLOCKED. SO THE CONCLUSION IS THAT THIS PROCESS THAT'S GOING ON DURING THIS PLASTICITY HAS SOMETHING TO DO WITH INVOLVING PROTEIN SYNTHESIS. THIS WAS ALL STAKING PLACE IN THE DEVELOPING SERVICE SYSTEM. IN HAPPENS IN ALL OF US IF YOU HAVE A MATURE SYSTEM OR NOT. THERE ARE PEOPLE STUDYING THE EFFECTS OF SLEEP ON MEMORY CONSOLIDATION IN THE MATURE NERVOUS SYSTEM AND A LOT OF THIS COMES FROM THE WORK OF ROBERT STICKLE AND WALTER AND COLLEAGUES. THIS IS A SIMPLE SLIDE BUT THEY USED THIS TEST CALLED THE TEXTURE DISCRIMINATION PATH WHICH I'LL EXPLAIN A LITTLE BIT MORE AS WE GO ON. BUT THE POINT HERE IS THAT WITH THIS TEST, IF THEY TRAIN THEM ON THE TASK, LET THEM SLEEP AND THEN TEST THEM AFTERWARDS THAT THEY DID BETTER THE LONGER THEY SLEPT. SO THIS IS SOMETHING THAT ALL STUDENTS SHOULD BE SHOWN. DON'T STAY UP ALL NIGHT AND STUDY FOR A TEST. GO STUDY ALL ALONG AND SLEEP THE NIGHT BEFORE. SO WE'RE VERY INTERESTED IN A COMBINATION OF THESE ISSUES. WE'RE INTERESTED IN PLASTICITIES IN PARTICULARLY THE ADULT'S BRAIN. AND WE'RE INTERESTED IN HOW PROTEIN SYNTHESIS IS INVOLVED IN THAT PROCESS DURING SLEEP. I'LL TELL YOU ABOUT A TEST THAT WE'RE DOING. WE ONLY HAVE PRELIMINARY RESULTS. THIS IS A -- RESEARCH AND -- AT NINDS WHO ARE BOTH SLEEP EXPERTS. KATHLEEN SCHMIDT -- AND WHO ARE PHYSICIANS WHO ARE WORKING ON THIS. AND MANY OTHERS ARE HELPING WITH IT. THESE ARE KIND OF PRIMARY PEOPLE. WE ARE ASKING THE QUESTION IS THE SLEEP DEPENDENT PERFORMANCE ON THAT DISCRIMINATION TEST ACCOMPANIED BY CHANGES IN PROTEIN SYNTHESIS IN THE PARTS OF THE BRAIN THAT ARE UNDER GOING THIS PLASTIC CHANGE. SO WORE STUDYING HEALTHY VOLUNTEERS. THEY'RE YOUNG PEOPLE, THERE ARE PEOPLE WITH NO NEUROLOGICAL OR PSYCHIATRIC DISEASES. AND WELL THERE'S NO SLEEP ISSUES AND PEOPLE WHO DON'T HAVE PROBLEMS SLEEPING MIGHT NOT HAVE PROBLEMS SLEEPING WHILE WE'RE DOING A PET SCAN. SO I HAVEN'T MENTIONED PET YET BUT THAT'S HOW WE MEASURE PROTEIN SYNTHESIS AND THAT'S HOW WE DO THIS STUDY. WE'VE DEVISED A WAY OF MEASURING PROTEIN SYNTHESIS WITH POSITRON EMISSIONS TOMOGRAPHY IN HUMAN SUBJECTS. THAT INVOLVES INJECTING A RADIO LABELED TRACER AMINO ACID AND THEN THROUGH KINETIC MODELING AND THE DYNAMIC SCAN PROCESS. WE ACTUALLY CAN COME UP WITH A RATE OF INCORPORATION OF THE AMINO ACID DO PROTEIN IN ALL PARTS OF THE BRAIN. SO OUR PROCEDURE, BECAUSE THEY HAVE TO SLEEP DURING OUR PET SCAN, WE HAVE TO SLEEP DEPRIVE THEM BEFORE THEY DO THE STUDY. AND THEN WE TRAIN THEM ON THE DISCRIMINATION TEST IN THE MORNING AND THEN THEY GET A PET SCAN WITH OR WITHOUT SLEEP SO WE HAVE ONE COHORT THAT SLEEPS, ONE COHORT THAT WE CONTINUE TO KEEP AWAY AND THEN RETESTED ON THE DISCRIMINATION TEST. SO JUST TO SHOW YOU A LITTLE MORE ABOUT THAT TASK, SO THE COMPUTER TASKS SO THEY'RE WALKING A SCREEN. THE FIRST SCREEN HAS CROSS HAIRS IN THE CENTER AND THEY ARE TOLD TO FOCUS ON THE CENTER OF THE VISUAL FIELD AND WATCH THAT CROSS HAIR. THEN THEY GET A BLANK SCREEN AND THEN THEY GET THE TEST SCREEN. SO THESE ARE SOME EXAMPLES OF TEST SCREENS. AND THE TEST SCREEN HAS A LETTER, L OR T IN THE CENTER OF THE VISUAL FIELD AND THEY'RE TOLD THAT THEY HAVE TO IDENTIFY WHAT THAT LETTER IS. AND THEN WE HAVE THESE LINES AND HERE THESE THREE LINES ARE ORIENTED VERTICALLY IN HERE, AND ORIENTED HORIZONTALALLY. HERE THEY'RE ON THE LEFT AND HERE THEY'RE ON THE RIGHT. WHAT WE WANT THEM TO DO IS TELL US WHAT'S THE LETTER IN THE CENTER OF FIELD AND ARE THESE LINES ORIENTED VERTICALLY OR HORIZONTALLY. SO THEY SEE THAT SCREEN FOR A VERY SHORT TIME AND THEN THEY GET THIS SCREEN WHICH IS JUST A MASS SCREEN AND THEN A BLANK SCREEN AND THEN THEY ANSWER, THEY PUT IN THEIR ANSWER. WHAT VARIES IS THE INTERVALS BETWEEN THESE TWO. AND IN THE BEGINNING, THEY GET ALONG, LONGER INTERVAL AND IT GETS PROGRESSIVELY SHORTER AND WE LOOK AT THAT TO DECIDE THEIR PERFORMANCE. SO JUST AGAIN TO EXPLAIN THE EXPERIMENT TO GO BACK TO THIS DIAGRAM, SO WE CAN SEPARATE THE TWO HEMISPHERES. WE CAN MEASURE PROTEIN SYNTHESIS IN EITHER, IN BOTH HEMISPHERES. IF WE ALWAYS PUT THE STIMULUS IN THE LEFT VISUAL FIELD, WE WILL BE ADDRESSING THE RIGHT HEMISPHERE. AND IF WE PUT IT IN THE RIGHT VISUAL FIELD THAT'S HERE, WE WILL BE ADDRESSING THE LEFT HEMISPHERE. SO WE HAVE AN INTERNAL CONTROL. SO THIS JUST SHOWS YOU THE PET SCAN SO WE DO OUR SCANS ON EITHER LATENT SLEEPING SUBJECTS, THEY ARE MONITORED FOR POLY SOME MOGRAPHY WITH THAT TWO AND-A-HALF MILLIMETERS. SO TO START THE SCAN ONCE THE SLEEPING SUBJECTS HAVE GONE TO SLEEP WE INJECT THE RADIO LABELED TRACER WHICH IS ONE C11 LUCIEN AND WE FOLLOW ITS DISAPPEARANCE IN THE ARTERIAL BLOOD OVER THE WHOLE COURSE OF THE SCAN. AT THE SAME TIME WE'RE SCANNING THE BRAIN AND YOU CAN SEE THE TRACER IN THE BEGINNING IS VERY LOW. IT'S TAKEN TO THE BRAIN AND THEN CLEARED. WHAT'S LEFT IS MOSTLY WHAT'S IN PROTEIN. AND WE VIEW AN MRI, WE GET THE ANATOMY AND THEN WE CAN IDENTIFY ALL THE REGIONS OF THE BRAIN AND TRANSFER THOSE OUTLINES TO THE PET IMAGE AND GET THE RATE. SO FIRST WE WANTED TO KNOW WELL, IF WE SLEEP DEPRIVE THESE GUYS, CAN THEY DO THE TASK AND CAN THEY SHOW IMPROVEMENT AFTER A NAP. SO WE JUST COMPARE THE SLEEP DEPRIVED GROUP OF THE CONTROL GROUPED, SLEEP SATED GROUP AND THEY ALL HAD A NAP BETWEEN THE MORNING AND ADMINISTRATION OF THE TEST AND THE EVENING. AND I SHOULD BE SURE TO POINT OUT THAT IMPROVEMENT, WITH IMPROVEMENT, THIS SCORE GOES DOWN. SO THEY ALL SHOWED IMPROVEMENT. SO THAT WAS REASSURING. AND NOW IN OUR COHORT OF SUBJECTS THAT WE STUDIED SO FAR, WE CAN SEE THAT THOSE THAT DID NOT, WERE NOT PERMITTED TO SLEEP DURING THE PET SCAN, WERE NOT PERMITTED TO NAP, ACTUALLY DID WORSE ON THE SECOND ADMINISTRATION OF THE TEST. AND THOSE THAT DID GET A NAP, DID BETTER. AS I SAID, WE DON'T HAVE FINAL RESULTS BUT I JUST WANTED TO SHOW YOU A PICTURE OF WHAT RESULTS IN SOME OF THE SUBJECTS LOOK LIKE. SO THIS IS THE PRIMARY VISUAL CORTEX. THE -- GYRUS IS HERE, LEFT AND RIGHT. AND THE LINGUAL IS BELOW. SO THE LINGUAL IS RESPONDING TO THE UPPER PART OF THE VISUAL FIELD. THE CUNEATE IS LOWER ON THE RIGHT SO THE RIGHT VISUAL FIELD IN THE ACTIVATION, IT'S NOT AN ACTIVATION LIKE WE USUALLY THINK. THIS IS AN INCREASE IN PROTEIN SYNTHESIS IS OCCURRING IN THE LEFT HEMISPHERE. SO THIS IS THE RESULTS THAT WILL BE CONSISTENT THROUGHOUT, WE DON'T KNOW YET BUT THAT'S WHAT WE'RE, THAT'S WHAT WE'RE LOOKING FOR FOR SURE. IF THAT HOLDS UP, THAT WOULD BE A DIRECT DEMONSTRATION OF INCREASE IN PROTEIN SYNTHESIS DURING MEMORY CONSOLIDATION TASK. SO JUST TO SUM UP, I THINK I'VE TALKED A LITTLE TOO LONG, I'M SORRY, SUSAN. I WILL JUST KIND OF GO OVER. WE TALKED ABOUT DEFINITION, DISORDERS AND SLEEP DRIVES. AND THEN WE WENT INTO PLASTICITY AND MEMORY AND NOW OUR STUDY WHICH IS TO DETERMINE IF THERE IS A PROTEIN DEPENDENCY ASPECT TO THIS PROTEIN MEMORY. SO THANK YOU VERY MUCH. [APPLAUSE] >> YOU MENTIONED IN STAGES OF SLEEP [INDISCERNIBLE]. >> CORRELATION LIKE -- LEVELS, IS THAT WHAT YOU MEAN. >> IT CHANGES, YES I DON'T KNOW. DO YOU KNOW? >> IF I MAY ADD [INDISCERNIBLE]. THAT IS THE CORRELATE [INDISCERNIBLE] DECREASE DURING [INDISCERNIBLE]. BUT MAYBE YOUR QUESTION ALSO MIGHT HAVE BEEN GETTING -- >> YES, NEUROTRANSMITTERS [INDISCERNIBLE]. >> WOULD YOU LIKE TO ADDRESS THE QUESTION? >> SURE. THE QUESTION WAS WHAT NEUROTRANSMITTER CHANGES HAPPEN DURING IN THE BRAIN DURING SLEEP AND THAT'S REALLY UNDER A LOT OF STUDY RIGHT NOW. WE DO KNOW THAT [INDISCERNIBLE] I'LL TELL YOU ABOUT A MINUTE THAT DOPAMINE ENERGETIC LEVELS -- IT'S RATHER A HOT TOPIC IN SLEEP BECAUSE THINK ABOUT THE EXPERIMENTAL CHALLENGES INVOLVED. CAROLYN'S GOT A REALLY WONDERFUL SYSTEM TO MEASURE THESE LEVELS OF PROTEIN IN THE BRAIN. BUT HOW WOULD YOU MEASURE A NEUROTRANSMITTERS DURING SLEEP EVEN IF YOU'RE USING ANIMAL YOU'RE TALKING ABOUT YOU HAVE TO SACRIFICE THE ANIMAL. AREN'T YOU CHANGING THINGS WHILE YOU'RE DOING THAT. YOU HEARD OF PEOPLE MICROWAVING THEM, BOILING THEM AND FREEZING THEM IN NITROGEN SO IT'S NOT EASY TO DO. >> LET ME ASK YOU A GENERAL QUESTION ABOUT PET SCANNING AND MRI. SO YOU SHOW EVIDENCE OF ENHANCED PROTEIN SYNTHESIS IN A CERTAIN AREA OF THE BRAIN ASSOCIATED WITH VISUAL PHENOMENON. NOW THE VISUAL PHENOMENON IS AN INCREDIBLY COMPLICATED BUSINESS INVOLVING NOT JUST THE OPTIC SYSTEM AND THE GENETICULAL BODY AND FEEDING INTO THE WHOLE CORTEX. IT'S A NETWORK, IT'S NOT A CAMERA. HOW DOES THAT PLAY INTO THE FACT THAT YOU WOULD LOCALIZE THIS TO A VERY SPECIFIC AREA IN TERMS OF THAT'S WHERE THE PRIMARY EFFECT IS. MAYBE IT'S THE SECONDARY COMPARED TO THE WHOLE CASCADE OF EVERYTHING THAT'S HAPPENING. I I AM NAIVE ABOUT IT. >> SO THAT IS A VERY GOOD POINT AND OF COURSE IN OUR STUDIES THEY'RE FOCUSED ON THE PRIMARY VISUAL CORTEX BECAUSE THAT'S THE PLACE WHERE THERE'S CONSOLIDATION OF THE INFORMATION FROM THAT VERY SIMPLE TASK. THIS IS, REMEMBER, NOT A COMPLICATED VISUAL TASK, IT'S A PRETTY SIMPLE THING, LOCATING THE DISEASE LINES THAT ARE LIKE THIS AND LIKE THIS. SO WE'RE FOCUSED ON THAT AREA AND THAT ERA'S BEEN SHOWN ON MRI SCANS IS AN AREA THAT IS ACTIVATED BY THIS TASK. AND EVEN REACTIVATED DURING SLEEP AFTER TRAINING. SO THAT'S ANOTHER VERY INTERESTING OBSERVATION. BUT WE THINK WE'RE LOOKING AT A PARTICULARLY PERTINENT AREA. BUT WE'RE NOT LOOKING AT EVERYTHING IN THE VISUAL SYSTEM. >> [INDISCERNIBLE] DIFFERENCES DURING SAY REM SLEEP [INDISCERNIBLE]. THERE USED TO BE A DRUG CALLED QUALUDE BUT THEY TOOK IT OFF THE MARKET BECAUSE PEOPLE WERE -- I DON'T KNOW. >> I DON'T KNOW, DOES ANYBODY KNOW. >> SUSAN, DO YOU HAVE A COMMENT ABOUT QUALUDES. >> I THINK THIS GOES BACK TO THE QUESTION A LITTLE BIT ABOUT NEUROTRANSMITTERS. FIRST I THINK IT'S IMPORTANT TO MENTION THAT IN GENERAL THIS IS A VERY GENERAL STATEMENT I HAVE TO EMPHASIZE THAT IN ECHOING THE EARLIER COMMENT, THE EARLIER RESPONSE TO ABOUT NO TRANSMITTERS IN THAT GENERAL AREA. BUT DURING NON-REM SLEEP YOU GET PROGRESSIVE DECREASES IN SEROTONIN, NOR EPINEPHRIN AND THEN DURING REM SLEEP YOU HAVE THE [INDISCERNIBLE] OF THESE LEVELS OF NEUROTRANSMITTERS OR THE FIRING RATE. BOTH STUDIES HAVE BEEN MEASURED IN NON-REM SLEEP. AND THEN IN REM SLEEP, WHAT'S UNIQUE IS THEY STAY BUT YOU HAVE INCREASE IN ACETYLCHOLINE. SO YOU SAY WHAT DRUGS DO ACETYLCHOLINE RELEASE AND SURE ENOUGH THOSE DRUGS DO ININCREASE REM SLEEP. IF YOU GET SLEEP OR YOU INHIBIT IT THEN YOU GET DECREASES GENERALLY. >> OKAY. WE'LL HAVE TIME FOR MORE QUESTIONS AFTER DR. HARBISON. SUSAN. >> DID EVERYBODY HERE ME OKAY. THANK YOU FOR THE OPPORTUNITY TO SPEAK TO YOU TODAY. WE'RE GOING TO DO SOMETHING A LITTLE BIT DIFFERENT. WE'RE GOING TO TALK ABOUT GENETICS AND DROSOPHILA. AS CAROLYN VERY NICELY POINTED OUT, THERE ARE A LOT OF -- OF CURBING YOUR SLEEP OR DEPRIVING YOURSELF OF SLEEP FULLY AND SLEEP DISORDERS ALSO LEAD TO PROBLEMS WITH YOUR HEALTH. SO FOR EXAMPLE, SHE MENTIONED THAT CURBING YOUR SLEEP CAN LEAD TO A HIGHER INCIDENCE OF OBESITY, DIABETES AND CARDIOVASCULAR DISEASE AND SLEEP DISORDERS LIKE INSOMNIA, A HALLMARK OF DEPRESSION AS WELL AS SLEEP APNEA BEING A RISK FACTOR FOR HYPERTENSION. SO WE WANT TO KNOW HOW SLEEP IMPACTS HUMAN HEALTH BUT WE REALLY DON'T UNDERSTAND THE FUNCTION OF SLEEP. THERE'S SOME CHALLENGES IN HUMAN SLEEP STUDIES. ONE IS SIMULTANEOUS MEASUREMENTS ARE ONLY POSSIBLE IN GENO TYPES IF WE HAVE MONO ZYGOTIC. YOU CAN'T FOR EXAMPLE CONTROL THE ENVIRONMENT THAT PEOPLE EXPERIENCE. WE ALL EXPERIENCE DIFFERENT ENVIRONMENTS AND YOU PROBABLY ARE VERY WELL AWARE THAT SLEEP, YOUR ENVIRONMENTAL CONDITIONS CAN AFFECT YOUR SLEEP. YOU SLEEP A LOT DIFFERENTLY THE NIGHT BEFORE YOUR THESIS PRESENTS OR YOUR VACATION ON THE BEACH FOR EXAMPLE. SO IT'S SORT OF EXTREME BUT YOU CAN UNDERSTAND WHEN YOU'RE DOING A HUMAN STUDY AND YOU'RE SAMPLING NUMBERS OF THE POPULATION, YOU COULD BE GETTING ONE AT ONE OF THE GOOD TIMES AND ONE OF THE BAD TIMES AND THERE'S NO REAL WAY TO KNOW THAT. THEN FINALLY ONCE YOU COME UP WITH GENES -- VERY DIFFICULT TO TEST. IT'S HARD TO TEST THEM IN HUMANS BUT THEY MAY NOT BE IN A GENERAL POPULATION OR NOT IN HIGH FREQUENCY DEFINITIVELY CLAIM A CERTAIN GENE AFFECTS THIS OR NOT. SO THAT'S WHERE DROSOPHILA HAS SOME SIGNIFICANT ADVANTAGES. THAT'S BECAUSE WE CAN TAKE POPULATIONS OF GROUPS OF FIVE AND INBREED THEM COMPLETELY AND THIS INBREEDING ALLOWS US TO CREATE GENO TYPES THAT ARE BASICALLY IDENTICAL WITHIN A LINE OF FLIES. EVERY FLY YOU TEST IN THAT LINE HAS THE SAME BASIC PHENOTYPE AND YOU CAN MEASUREMENTS SIMULTANEOUSLY. SO THAT'S A REAL ADVANTAGE. WE CAN COMPLETELY CONTROL THE ENVIRONMENT. I'VE GOT TEMPERATURE -- THE FOOD, THE MATING STATUS, THEIR CROWDING, HOW THEY HAVE IN THEIR BIORHYTHMS. WE HAVE GENETIC TOOLS IN DROSOPHILA. THERE ARE TOOLS LIKE THE -- UAS SYSTEM THAT ALLOWS US TO DRIVE -- WITH A ALSO HAVE MUTATIONS AND TRANSGENICS AS WELL AS DELETIONS THAT CAN BE TESTED. SO YOU HAVE A REGION OR GENE THAT'S PRETTY EASY TO JUST ORDER SUPPLIES FROM THE DROSOPHILA STOCK CENTER AND THEN TEST YOUR MUTANTS. SO THE CHARACTERISTICS OF SLEEP AND I'LL JUST TOUCH ON THEM BRIEFLY HERE. SHE MENTIONED AN INCREASED ARRIVAL THRESHOLD. THAT'S A REDUCED RESPONSE TO EXTERNAL STIMULI. HOMEOSTATIC DEPRIVATION. YOU WANT TO MAKE UP FOR THE SLEEP YOU LOST. THIS IS SPONTANEOUS AND REVERSIBLE. IT SEEMS TO BE RELATIVELY STABLE CHARACTERISTIC WITH AN A SPECIES. SO A GROUP SHOWED THAT [INDISCERNIBLE] HAVE THESE BEHAVIORAL CHARACTERISTICS OF SLEEP. INCLUDING SLEEPING IN THE BED. AND SO AS FAR AS IT CAN BE USED A POWERFUL MODEL ORGANISM FOR THE STUDY OF MAMMALIAN SLEEP. SO WE MEASURE SLEEP IN FLIES USING THE DROSOPHILA ACTIVITY MONITOR OR GLASS TUBES THAT FIT INTO THESE LITTLE HOLES. THERE'S A SINGLE FLY IN THOSE GLASS TUBES SO FOR EACH MONITOR THEY MEASURE 31 OR 32 OF THESE. THE MONITOR HAS AN INFRARED BEAM THAT BISECTS IT SO WHEN THE FLY WALKS BACK AND FORTH [INDISCERNIBLE] RECORDED ON THE COMPUTER. WHEN YOU GET A DATA FILE WHICH GIVES YOU A NUMBER OF ACTIVITY COUNTS PERMANENT OF RECORDED TIME. AND I DON'T HAVE TIME TO REALLY GO INTO WHY THIS IS BUT YOU CAN ASK ME THE QUESTION FIVE MINUTES WITHOUT AN ACTIVITY COUNT IS DEFINED AS -- AND THAT'S DIFFERENT FROM JUST A TRANSIENT REST PERIOD AS ONE MINUTE. SO IF WE HAVE ALL OF THESE MEASURES FOR THE FLIES, WE CAN CALCULATE THE DURATION, THE NUMBER OF SLEEP OUTS OR ANALYSIS THE FLY TAKES, THE AVERAGE DURING THE DAY OR NIGHT AND THE WAKING ACTIVITY WHICH IS THE NUMBER OF ACTIVITY COUNTS BEING PROCESSED OVER TIME. TO APPROACH THE STUDY OF SLEEP USING SYSTEMS GENETICS WE START FIRST OF ALL WITH A WILD DERIVED -- POPULATION AND I'LL TELL YOU ABOUT OUR POPULATION IN A MINUTE. THE MEASURE OF THE SLEEP PHENOTYPES ON THESE FLIES AND THEN WE OBTAINED DNA SEQUENCE FOR ALL OF THE INDIVIDUAL LINES IN THE POPULATION. SO WE MAY HAVE SINGLE POLYMORPHISMS, YOU MAY HEAR ME REFER TO THEM. LET'S A SINGLE POLY MORPHIC DIFFERENCES. WE MEASURE INSERTIONS AND DELETIONS AND CHARACTERIZE THEM AS WELL THAT MAY BE PRESENTED. WE MEASURE THE LEVEL USING THE RNA. THEN WE CAN ASSOCIATE OUR DNA SEQUENCE POLYMORPHISMS WITH OUR PHENOTYPES IN ORDER TO IDENTIFY QUANTITATIVE -- WE CAN ALSO REGRESS THE -- AND THEN WE CAN ALSO ASSOCIATE THESE POLYMORPHISMS WITH THE TRANSCRIPT ABUNDANCE TO IDENTIFY EXPRESSION QUANTITATIVE. SO THESE ARE THE POLYMORPHISMS THAT INFLUENCE THE TRANSCRIPT ABUNDANCE. I'LL TALK ABOUT THE FIRST ONE HERE WHICH IS -- WITH THIS PHENOTYPE. WE USE THE DROSOPHILA PANEL -- FATING FOR 20 GENERATIONS SO EACH LINE OF FLIES WHICH I HAVE HERE IS A SOLID COLOR HAS AN IDENTICAL GENOTYPE. BUT BETWEEN LINES THERE'S A LOT OF GENETIC VARIATION AND THAT'S AN IDEAL SCENARIO FOR A PHENOTYPE. JUST FOR THE SAKE OF TIME I'M GOING TO FOCUS ON NIGHT SLEEP AND DAY SLEEP. THAT'S THE DURATION. I'LL SAY NIGHT SLEEP OR DAY SLEEP AND TALK ABOUT THE DURATION IN MINUTES -- SO IF WE LOOK AT A HISTOGRAM OF THE DROSOPHILA GENETICS PANEL -- WE CONSIDER THERE'S AN ENORMOUS AMOUNT OF VARIATION. SO IF YOU THINK ABOUT THE NIGHT'S PERIOD, 12 HOURS. SO THAT'S ZERO 720 MINUTES. WE HAVE A LINE AROUND 90 MINUTES FEMALES AND THESE MALES ARE OVER 11 HOURS. SO IT'S QUITE EXTREME. THE ABILITY HERE IS QUITE HIGH INDICATING THAT THERE'S A LARGE GENETIC COMPONENT FOR THIS PARTICULAR TRAIT. THOSE ARE HIGHLY VARIABLE. YOU NOTICE THE DIFFERENCE IN THE PATTERN OF THE HISTOGRAMS HERE, THE FEMALES IS A LOT LESS DURING THE DAY THAN THE MALES. THE MALES SLEEP A LOT MORE. WE OFFER SOMETHING THAT WAS REALLY INTERESTING IN OUR LINES OF FLIES. WE COUNTED EVERY FLY IN THE EXPERIMENT SO THAT THE FEMALES ARE SHOWN IN PINK AND THE MALES ARE IN BLUE. THIS IS EVERY FLY. THE HOLES ON THE LINES HERE ARE EACH A LINE OF FLIES FROM THE DROSOPHILA GENETIC REFERENCE PANEL. THERE ARE 168 OF THESE. AS YOU CAN SEE HERE IF WE ORDER THIS FROM LONGER SLEEPING TO SHORTER SLEEPING FOR THE NIGHT SLEEP AND THE DAY SLEEP, WE SEE THAT THE LONG SLEEPERS ARE FAR LESS VARIABLE THAN THE SHORT SLEEPERS. WHICH IS PRETTY REMARKABLE. AND THAT SAYS A LOT ABOUT HOW MUCH VARIATION YOU CAN GET IN FLIES WHO ARE IDENTICAL GENO TYPES. YOU CAN SEE THESE GUYS REALLY ARE QUITE EXTREME IN THEIR VARIATION. THE SLEEP IS QUITE LOW. WE CAN CALCULATE THIS PARTICULAR LEVEL OF VARIATION OF THE COEFFICIENT OF ENVIRONMENTAL VARIATION OR CVE. THAT IS THE STANDARD DEVIATION -- IF YOU HAVE A LINE HERE WHICH IS CONSOLIDATED I GUESS YOU WOULD SAY ITS PHENOTYPE THEN YOU CAN SEE THE CVE IS RELATIVELY LOW. THOSE FLIES ARE ROBUST IN PERTURBATION. IF YOU LOOK HERE AT THE SHORT SLEEPERS, THEY HAVE A BROAD RANGE, THEIR CVE IS HIGHER AND THEY ARE SENSITIVE TO ENVIRONMENTAL CHANGES. SO IT TURNS OUT THAT THIS COEFFICIENT VARIATION IS -- YOU CAN SEE THAT THE MALE HERE IN THIS PARTICULAR GROUP ARE RELATIVELY SENSITIVE AND SO ARE THE FEMALES. WE HAVE SOME VERY SENSITIVE FEMALES OUT HERE. THEY'RE SENSITIVE TO CHANGES IN THE ENVIRONMENT. THE SAME IS TRUE TO -- WITH THE MALES HERE BEING A LITTLE BIT LESS SENSITIVE THAN FEMALES. AND THAT MIGHT EXPLAIN WHY THEY'RE ABLE TO SLEEP MUCH DURING THE DAY. THEY'RE RELATIVELY INSENSITIVE TO DIFFERENT ENVIRONMENTAL CHANGES LIKE THE LIGHT CENTERS FOR EXAMPLE. IT TURNS OUT THESE MEASURES ARE CORRELATED. IF YOU CORRELATE NIGHT SLEEP WITH THE RESPECTIVE COEFFICIENT OF THE VARIATION, THE CORRELATIONS HERE ARE PRETTY HIGH. FROM THAT TELLS US IS THAT IF YOU FIND A POLYMORPHISM [INDISCERNIBLE] VARIATION IN NIGHT SLEEP AS WELL. THE SAME FOR DAY SLEEP. SO WE DID A GENOME-WIDE ASSOCIATION ON THESE FLIES. I'M JUST SHOWING YOU KIND OF A CARTOON HERE. JUST TO REMIND YOU THAT EACH OF THE LINES ARE HOMOZYGOTE. THIS IS ANOTHER THING THAT MAKE THE ANALYSIS OF THESE COMPLAD THAN IN THE HUMAN SITUATION WHERE YOU SEE A LOT OF HETEROZYGOTE. YOU SEE NO ASSOCIATION IF THERE'S A SLEEPER OF A PARTICULAR GENOTYPE HAVE A PARTICULAR GENOTYPE AND SHORT SLEEPERS HAVE A PARTICULAR GENOTYPE THAT WOULD BE A PARTICULAR ASSOCIATION. THERE ARE POLYMORPHISMS IN THE REFERENCE PANEL THAT'S BEEN FULLY SEQUENCED. HERE I'M JUST SHOWING YOU THE DISTRIBUTION. THESE COLORS HERE REPRESENT THE FIVE CHROMOSOME ARMS OF DROSOPHILA, AND THEN THIS IS THE COUNT IN MILLIONS OF SITES FOR ENERGETIC ELECTRONIC POLYMORPHISMS AND THESE ARE POLYMORPHISMS THAT ARE SYNONYMOUS -- WHAT THAT MEANS IS THAT THESE GUYS ARE IMAGING AND THESE GUYS ARE OUTSIDE OF IT. YOU CAN SEE THE POLYMORPHISMS OUTSIDE THE GENES ARE FAR MORE NUMEROUS THAN THE ONE INSIDE THE GENES. AND OF COURSE WE EXPECT THAT DUE NATURAL -- MORPHISM INSIDE THE -- OF GENES. SO THERE'S 2,490,165 POLYMORPHISMS. THAT WE ASSOCIATED A SLEEP PHENOTYPES WITH. WE FOUND THAT MANY SNPs ARE SIGNIFICANTLY ASSOCIATED WITH NIGHT AND DAY SLEEP. HERE I'M GOING TO EMPHASIZE NIGHT SLEEP. THE VERIFICATION ON THOSE SO I'LL EMPHASIZE THOSE IN MINUTE. WE FOUND 160 SNPS -- I MEAN ABOUT 5% MINOR LOW FREQUENCIES. THE POPULATION HAD THE MAJOR -- SO MOST OF OUR SNPs ARE THESE KIND OF LOW FREQUENCY SNPs. THERE ARE 18 HUMAN -- FOR NIGHT SLEEP AND 43 FOR NIGHT SLEEP CVE. THE DAY SLEEP AT OUR THRESHOLD LEVEL WE DIDN'T FIND ANY SNPs WHICH IS VERY DEPRESSING. FOR DAY SLEEP CVE WE FOUND 71SNPs IN 23 CANDIDATE GENES. THESE ARE PREDOMINANTLY -- HOMOLOGUES. SO MOST OF OUR SNPs OVERLAP BETWEEN NIGHT SLEEP AND NIGHT SLEEP CVE. IN FACT ALL BUT ONE GENE PRETTY MUCH OVERLAPPED BETWEEN THESE TWO TRAITS. SO THAT WE REFLECT THAT HIGH GENETIC CORRELATION BETWEEN THE TWO TRAITS THAT I MENTIONED BEFORE. SO THAT'S WHAT WE EXPECTED TO FIND. AND A LOT OF THESE SNPs ARE FOUND IN HOMOLOGUE OF HUMAN SLEEP GENES. SO THERE WAS A GENOME-WIDE ASSOCIATION STUDY DONE BY -- WHO WAS THE FRAMINGHAM POPULATION. HE LOOKED AT DAY TIME SLEEPINESS. USUAL BEDTIME IS SLEEP DURATION AND THIS IS DONE WITH A DIARY ASSAY WHERE THE CORE BASICALLY YOUR SLEEP PATTERNS OVER A PERIOD OF ABOUT SEVEN DAYS. AND WE HAVE FOUR GENES THAT OVERLAP WITH GENES WE FOUND IN THAT STUDY. AND THERE'S ALSO BEEN STUDIES IN NARCOLEPSY AND SLEEP APNEA WHERE WE FOUND SOME OF OUR GENES OVERLAPS. SO THIS IS QUITE PROMISING BECAUSE IT SUGGESTS A GENETIC BASIS OF SLEEP BETWEEN SIZE AND HUMANS. SO I JUST SHOWED YOU A LOT OF SNPs THAT WERE ASSOCIATED WITH SLEEP. IN HUMAN STUDIES, THIS IS WHERE THE REAL DIFFICULTY BEGINS. YOU MIGHT FIND SOME SNP THAT YOU THINK ARE INTERESTING BUT HOW DO YOU GO ABOUT PROVING THEM. WITH SLIDE WE CAN TAKE AN APPROACH TO CONFIRM THAT THE SNPs FROM OUR GENOME WIDE STUDY. WE HAVE TO START WITH A LARGE POPULATION OF FLIES. I'LL SHOW YOU HOW WE CREATED THIS. WE CAN SELECT FOR SHORT SLEEPER, FOR LONG SLEEPER -- AND SO WE'RE GOING TO MEASURE SLEEP IN EACH GENERATION TO DO THAT SELECTION. BUT OF COURSE WE WANT TO KNOW WHAT'S HAPPENING TO THE UNDERLYING GENOTYPES. AND THEN ONCE WE GET THE GENO TYPIC -- THAT WILL ALLOW US TO IDENTIFY THEM. THAT'S THE HOPE. SO WE BEGIN WITH THE TEN MOST EXTREME -- LINES. ON THE LEFT HALF OF THE GRAPH OUR SHORT SLEEPERS. WE'RE GOING TO DO THE SELECTION ON NIGHT SLEEP. THESE ARE THE REDDISH PURPLE BARS THAT SHOWED YOU NIGHT SLEEP. WITH THAT DAY SLEEP IN THE COLOR IN 24 HOUR SLEEP IN SOCIAL OF BLUISH PURPLE COLOR. THAT'S JUST TO HE SHOW YOU THE CONTRASTED DIFFERENCE. SO EVEN IF YOU LOOK AT THIS HOUR SLEEP THE SHORT SLEEPERS ARE LESS THAN THE RIGHT SIDE OF THIS GRAPH ARE THE LONG SLEEPERS. AND THESE GUYS HERE ARE ALMOST UNIFORMLY SLEEPING ABOUT 11 HOURS AT NIGHT ON AVERAGE. SO THEY ARE LONG SLEEPERS. YOU TAKE THOSE LINES AND I'M JUST SHOWING YOU KIND OF A CHROMOSOME SCHEMATIC OF THE SLIDES. AND THE LONG SLEEPER SNPS ARE IN THE BLACK COLOR AND THE LONG ONES ARE IN YELLOW. YOU CAN CROSS THESE TOGETHER. SO WE TAKE THE MALES OF EVERY LINE AND CROSS THEM WITH THE FEMALES OF EVERY OTHER LINE. AND WE CAN TAKE THE PROGENY OF THAT AND THEN ALLOW THEM TO MATING TO OCCUR. WE DID THIS FOR 21 GENERATIONS IN SIZE. THAT'S EACH GENERATION IS ABOUT TWO WEEKS, CLOSE TO A YEAR BUT JUST ALLOWED THESE GUYS TO MIX. THIS GIVES US A HIGHLY VARIABLE MIX POPULATION. WE KIND OF SEE WHAT WE'RE DOING NOW. WE'VE TAKEN THE EXTREME LINES AND WE'RE JULY -- JUMBLING UP THOSE AND SEE WHAT WE GET OUT IN THE END. IT'S A NICE WAY TO EPICATE POPULATION. SO WE SPLIT THIS MIXED POPULATION INTO THE REPLICATE POPULATIONS. WE HAD TWO LONG SLEEPER POPULATIONS, TWO SHORT SLEEPER POPULATIONS AND TWO CONTROLLED. WE MEASURE NIGHT SLEEP AND A HUNDRED MALES AND A HUNDRED FEMALES FROM EACH POPULATION. AND THEN WE USE THE LONGEST 25% OF THE NEXT GENERATION FOR THE LONG SLEEPERS. THE SHORTEST SLEEPING -- NEXT GENERATION FOR THE SHORT SLEEPERS AND FOR THE CONTROL WE JUST CHOSE 25% AT RANDOM. SO WE'RE LOOKING HERE AT THREE CHARACTERISTICS IN OUR REPLICATE POPULATIONS SELECTION. THE ONLY THING I WANT TO SHOW YOU HERE IS THAT FOR OUR CONTROL, A LONG SLEEPER AND OUR SHORT SLEEPER BUT THERE WASN'T ANY DIFFERENCE BECAUSE IT'S SOMETHINGANT AND THAT'S GOOD. WE HAVE NO BIAS IN THE POPULATION. NOW LET'S LOOK AT THE GENOTYPING BECAUSE THAT'S REALLY WHAT WE WANT TO GET TO. SO WE DID THIS A COUPLE WAYS. I'M GOING TO TELL YOU ABOUT THE WAY WE'RE GOING TO HAVE DATA AND THEN I'LL TELL YOU AT THE END THE OTHER DATA WE'RE LOOKING AT THAT. WE FIRST CHOSE MARKER LOCATIONS THAT WERE RELATIVELY EQUALLY SPACED ON THE FIVE CHROMOSOME LINES OF DROSOPHILA. WE TRIED TO PICK SNPs FOR GENES. THAT WAS FOR A -- ASSAY WHICH IS A PCR-BASED ASSAY. AND I'M GOING TO DESCRIBE THAT IN A MINUTE. SO THIS JUST SHOWS YOU DISTRIBUTION OF MARKER ALLELES IN OUR MOST EXTREME LINE. SO AGAIN OUR LONG SLEEPERS ARE AT THE TOP AND OUR SHORT SLEEPERS ARE AT THE BOTTOM. IF WE LOOK AT THE FLIES THAT HAVE THE MAJOR ALLELE VERSUS THE MINOR ALLELE, WE CAN SEE THAT THE LONGER SLEEPERS TEND TO HAVE THE MAJOR ALLELE AND THE SHORT HAVE THE -- RELEVANT IN A SECOND ALSO. BUT TO DO THE TAQMAN ASSAY, YOU CAN DESIGN DNA PROBES THAT WILL AWE -- TO YOUR SNPS. THE MAJOR ALLELE A AND WE HAVE THE PROBE -- AMEAL TO THIS PARTICULAR A. THIS WILL AMEAL TO THE MINOR ALLELE. EACH OF THESE PROBES HAS A FOUR FLOOR ON IT. THIS ONE HAS A GREEN -- AND IT HAS A CLENCHER ON IT ALSO. THAT MEANS THAT WHEN YOU PUT THESE PROBES INTO A SOLUTION, YOU DON'T REALLY HAVE ANY FLORESCENCE. AND THEN AS TTR PROCEEDS, THIS PROBE SPECIFICALLY TO YOUR SNPs. YOU'VE GOT -- HERE TOO AND HOPEFULLY WHAT YOU SEE IS A LOT OF GREEN FLORESCENCE IN THIS CASE. WE DO THIS ON A GRAND SCALE WHICH WAS FLUORODINE. THIS ALLOWS TO PUT INDIVIDUAL FLIES INTO ONE SIDE OF THE CHIP AND OUR TAQMAN ASSAY FROM 96 ON TO THE OTHER SIDE. THESE WELLS HERE, EACH ONE IS 9 MILLILITERS. WHAT WE GET OUT OF IT IS ABOUT 10,000 GENOTYPES. AND WE CAN PICK OUT THE HOMOZYGOUS MAJOR, HOMOZYGOUS MINOR AND HETEROZYGOUS GENOTYPES. SO THIS IS WHAT OUR ALLELE SEQUENCES WERE PRIOR TO SELECTION IN THE DGR PEAK WHY -- AND IN OUR MIXED POPULATIONS. I'M JUST SHOWING YOU, I DON'T HAVE MY NUMBERS HERE BUT ON THE Y AXIS IT REALLY GOES FROM ZERO TO A HUNDRED PERCENT IN 20% INCREMENTS. YOU CAN SEE THE VAST MAJORITY OF THE EACH OF OUR SNPs WHICH IS VERTICAL LINES IS THE MAJOR ALLELE. THAT'S WHY IT'S THE MAJOR ALLELE AND THE BLUE LINES OF THE ALLELES TAKE UP THE REST. THEY ARE SIMILAR BETWEEN THE PARENTAL LINES AND THAT INITIAL POPULATION. BUT OF COURSE IN THIS POPULATION IF WE LOOK AT THE GENOTYPES NOW, I'M SHOWING THE HOMOZYGOUS MAJOR IN GREEN AND THE HOMOZYGOUS MINOR IN BLUE AND THE MINOR IN PURPLE AND OF COURSE WE HAVE MORE HETEROZYGOUS. NOW WE HAVE TO DEAL WITH HETEROZYGOUS. THAT'S NOT EASY ANYMORE. SO LET'S LOOK AT A PHENOTYPIC RESPONSE TO SELECTION. NIGHT SLEEP SELECTION HERE SHOWS YOU THE GENERATIONS FROM -- 12 SO WE HAVE 12 GENERATIONS OF SELECTION. THEY SLEEP A LOT LONGER. THE SHORT SLEEPER POPULATION SLEEPS A LOT LESS AND THE CONTROL IS RELATIVELY FLAT. THE DAY SLEEPERS HAVE THE SAME TREND, THE LONG SLEEPERS SLEEP LONGER, THE SHORT SLEEPERS SLEEP LESS AND THE CONTROLS APPLY. I JUST WANT TO QUANTIFY THIS FOR YOU BECAUSE IT'S PRIVATE STRAIN. I WAS REALLY PLEASED WITH THE WAY THIS TURNED OUT. THE LONG SLEEPERS SLEEP ABOUT THREE AND-A-HALF HOURS MORE AFTER 12 GENERATIONS AND THE SHORT SLEEPERS SLEEP ABOUT SEVEN HOURS. SO ALWAYS GOT ABOUT TEN HOURS DIFFERENCE BETWEEN THE LONG SLEEPER AND SHORT SLEEPER LINES. SO THAT'S REMARKABLE. WITH DAY TIME SLEEPS, BECAUSE WE HAVE A POSITIVE GENETIC CORRELATION BETWEEN NIGHT SLEEP AND DAY SLEEP, WE -- CORRELATED RESPONSE TO SELECTION. SO DAY SLEEP DOES INCREASE AND NIGHT SLEEP DOES DECREASE ALTHOUGH NOT SO MUCH FOR THIS. WE SAW ANING TREATMENT DECREASE IN THE PHENOTYPIC VARIANCE AND WE SORT OF EXPECT THAT IN A SELECTION EXPERIMENT. YOU'RE BASICALLY -- TAKING A SMALL BIT OF THE POPULATION AND USING IT EACH TIME. SO YOU EXPECT THAT TO DECREASE AND IT DOES FOR A LONG LINE THAT I'M SHOWING YOU HERE. ALMOST ACCORDING TO 245E RENT CULL PREDICTION -- SOLID LINE. THE SHORT SLEEPERS HOWEVER DID NOT INCREASE SO MUCH. I THINK THAT'S A REFLECTION OF THAT VARIABILITY WE SAW EARLIER AMONG THE INDIVIDUAL FLIES. SO THE SHORT SLEEPERS ARE QUITE INTERESTING. WE DID SEE VERY LITTLE EVIDENCE HOWEVER OF INBREEDING. THIS OTHER GRAPH IS MEANT TO SHOW YOU, I'M A LITTLE NERVOUS HERE BECAUSE I'M RUNNING OUT OF TIME. WHAT I WANT TO HE SHOW YOU IS THAT NUMBERS OF FLIES THAT WERE VIABLE REALLY DIDN'T DO -- THROUGH GENERATIONS OF SELECTION EVEN THOUGH WE'RE INBREEDING. LET'S LOOK AT THE GENETIC RESPONSE TO SELECTION. THESE POPULATIONS CAN BE SUBJECT TO GENETIC DRIFT WHICH IS SIMPLY A RANDOM OF CHANGE IN THE GENETIC COMPOSITION OF OUR POPULATION OVER TIME. SO WE HAVE A RED HOMOZYGOTE, BLUE HOMOZYGOTE AND -- YOU CAN GET A DECREASE IN HOMOZYGOTE. LONG ENOUGH THE POPULATION GOES TO FIXATION AND IT DOESN'T MATTER WHAT FLY MATES WITH ANOTHER. YOU'RE NOT GOING TO CHOKE THIS. THE SMALLER POPULATION SIZE WE HAVE, THE MORE YOU EXPECT DROSOPHILA -- AND SO WE LOOK AT OUR MARKERS HERE. WE'RE IN GOOD SHAPE. WE DON'T REALLY EXPECT GENETIC DRIFT TO BE -- 12 GENERATIONS OF SELECTION AND I'M JUST TELLING YOU 12 SO WE'RE NOT QUITE AT THE POINT WE'RE LOOKING AT. SO THIS IS THE RESULTS THAT WE HAVE FOR GENOTYPES. THIS IS THE LONG SLEEPERS AT GENERATION ZERO, FIVE AND TEN. AND AGAIN I'M SHOWING YOU THE PERCENTAGES HERE OF THE MAJOR MINOR AND HETEROZYGOUS ALLELES OR GENO TYPES RATHER. AND WE CAN SEE HERE THAT THE LONG SLEEPERS TEND TO GET MORE HETEROZYGOUS. I THINK THAT'S A REFLECTION OF THE FACT THAT THOSE MAJOR ALLELES ARE MORE PREVALENT IN THE LONG SLEEPERS AS WELL AS POSSIBLE DOMINANTS. SO HERETO -- HETEROZYGOTE IS SIMPLY DOMINANT. IN SLEEPERS THE SITUATION IS A LITTLE BIT MORE STRANGE. WE ACTUALLY HAVE QUITE A FEW MORE GENOTYPES TO GO. BUT WE SAW QUITE A FEW MINOR ALLELES IN THE BEGINNING. THEY KIND OF WENT AWAY BY GENERATION FIVE AND BY GENERATION TEN THEY'RE COMING BACK. I CAN ONLY SAY WE CAN'T EXACTLY EXPLAIN THIS YET BECAUSE WE STILL HAVE ABOUT 137,000 GENO TYPES TO RESOLVE. I THINK WHAT THE NUMBERS HERE -- WE SAW SOMETHING SIMILAR IN THE CONTROLS. THE MINOR ALLELES DECREASED A LITTLE BIT AND THEN KIND OF CAME BACK IN GENERATION TERMS. SO IF YOU DO THESE CALCULATIONS -- WE FOUND SIX SNPs THEY WERE SIGNIFICANTLY DIFFERENT BETWEEN THE SHORT SLEEPER AND CONTROL AND THE LONG SLEEPER AND CONTROL. THOSE GENES FALL INTO THIS PARTICULAR GENE FOR A LONG SLEEP IT'S IN THE -- AND THE FUNCTION'S UNKNOWN BUT WE FOUND ALSO FOR SHORT SLEEP A SNP IN THE COATING REGION,LY SNPs -- ONE IS IN THE TRANSCRIPTION FACTOR AND ONE IS THE GENOME -- AND ONE IS INVOLVING ADHESION AND THEN ANOTHER GENE WHICH HAS AN UNKNOWN FUNCTION. THERE'S A SNP UP STREAM THAT'S ASSOCIATED. SO WE FOUND THESE LARGE CHANGES IN PHENOTYPES WITHIN A FEW GENERATIONS. IT'S REALLY REMARKABLE BUT IT SEEMS LIKE THE GENO TIMING IS A LITTLE BIT MORE SUBTLE. ONE WAY THE GENOTYPING USING THE -- -- WE ARE GOING TO LOOK AT THAT. WE ARE GOING TO LOOK AT OTHER SNPS IN THE GENOME. THE DIFFERENCE IS YOU CAN'T ASSOCIATE THE SNP TO A -- WE HAVE TO ASSOCIATE IT TO AN AVERAGE -- WE'RE FOLLOWING UP ON THAT ANALYSIS. AND -- ARE IMPORTANT HERE -- SO THAT'S EVERYTHING. I DO WANT TO THANK THE PEOPLE WHO ARE DOING THE WORK. -- SHE'S MY TECHNICIAN IN THE LAB AND SHE DID THE WORK OF THE SLIDE SHOW I JUST SHOWED YOU AND AMANDA DID THE VIABILITY STUDY. THANK YOU VERY MUCH. [APPLAUSE] >> THE HUMAN [INDISCERNIBLE]. >> THE HUMAN HOMOLOGUE ASSOCIATED WITH COOL CUT PHENOTYPES IN HUMANS. SO THESE, ARE YOU TALKING ABOUT [INDISCERNIBLE]. SO NO. THE POLYMORPHISMS AND THESE ARE CANDIDATE GENES. SEVERAL OF THESE ARE ACTUALLY IN BETWEEN GENES AND THESE ARE THE CLOSEST LOCI TO THEM, RIGHT. AND THERE'S MORE OR LESS THIS ONE IS ONLY -- SO THE CHANGE IN PROTEAN FUNCTION ISN'T NOBLE AT THIS POINT. IT DOESN'T MEAN IT'S NOT DOING ANYTHING. THAT'S WHAT MAKES IT MORE DIFFICULT IN HUMANS, YOU FIND THIS ASSOCIATION THAT'S STATISTICAL, RIGHT. SO YOU DON'T REALLY KNOW WHAT THE CAUSAL IS. THAT'S WHAT WE'RE TRYING TO DO. I SHOULD MENTION THAT YOU KNOW, IN HUMANS, WE'RE NOT LOOKING AT ALL OF THE GENE SEQUENCE, WE'RE LOOKING AT THE CHIPS THAT GENOTYPES ACROSS THE GENOME. SO WE'RE NOT GETTING EVERY SNP. THAT MEANS NECESSARILY THAT THERE'S PROBABLY BETWEEN THE CHIP AND THE ACTUAL CAUSAL SNP. IN FLIES THE SITUATION IS DIFFERENT. THERE'S VERY LITTLE LINKAGE IN FLIES. IT ACTUALLY IS OVER ABOUT 10 TO 30 BASE PAIRS WHICH IS UNBELIEVABLE. IF YOU FIND A SNP INSIDE A CANDIDATE GENE THAT SHOULD BE YOUR CAUSAL SNP. >> [INDISCERNIBLE]. >> OKAY, SO YOU'RE ASKING -- I'M SORRY, I'M NOT REPEATING. YOU'RE ASKING IF THERE'S A SNP ASSOCIATION IN SOME GENE WITH DAYME SLEEPINESS. THAT'S CORRECT. WE ALSO FOUND ONE IN THE SAME CANDIDATE GENE IN -- AND SO THAT HAS SOMETHING TO DO WITH THE ENERGY SIGNALING WHICH IS SIMULATED AT LEAST IN HUMANS -- THE RELATIONSHIP THERE. >> ARE THERE OTHER WAYS IN WHICH YOU CAN DEFINE SLEEPING OTHER THAN MOVING. FOR EXAMPLE YOU COULD BE SELECTING FOR OTHER PHYSICAL ACT DEFICIENCY RATHER THAN SLEEP. HOW DO YOU KNOW IT'S SLEEP AND NOT JUST MOTION THAT YOU'RE MEASURING? >> ONE OF THING WE DO IS WE LOOK AT OTHER PARAMETERS. ACTIVITY IS REALLY AN IMPORTANT ONE. IF WE CAN SHOW THE FLY'S MOVEMENT DURING THE DAY IS NOT RELATED TO SLEEP THEN YOU FEEL MORE CONFIDENT THAT THERE'S NOT ACTIVITY. IN THAT'S THE CASE THERE'S NO CORRELATION BETWEEN ACTIVITY AND SLEEP PARAMETERS -- SLEEP DURATION -- ANOTHER ASPECT. OH, EVERYBODY'S LOOKING FOR THAT. THERE ARE TIMES IN THE DISCUSSION THAT THE CORRELATES -- SO THE ONLY LIMIT I KNOW OF IS ONE THAT PAUL -- HAS COME UP WITH AND THAT'S AMYLASE. BOTH IN FLIES AND IN HUMANS. THERE'S UP REGULATION OF AMYLASE WHEN YOU'RE AWAKE AND THEN IT GOES DOWN IN YOUR SLEEP. THE SAME THING HAPPENS WITH HUMANS. THEY SHOW THAT DURING AWAKE OR SLEEP DEPRIVATION THE LEVELS OF AMYLASE IN A LOT OF HUMANS ARE UP AND THEN IT COMES DOWN. THERE'S ONE POTENTIAL BIOMARKER THAT EVERYONE'S LOOKING FOR A BIOMARKER THE TYPE YOU DESCRIBED. >> [INDISCERNIBLE] ARE THEY DEFINING SLEEP BY WALKING UP AND DOWN THE TUBE OR SOME OTHER WAY. >> IN HUMANS? IN THE FLIES. SO FOR THAT PARTICULAR STUDY, YES, THE SAME WAY IN THE FLIES. >> [INDISCERNIBLE] >> I HOPE SO. >> ANY OTHER QUESTIONS? >> SO I GATHER WHAT YOU'RE TRYING TO DO IS YOU'RE TRYING TO IDENTIFY GENETIC MARKERS IN THE HUMAN GENOME THAT THEN YOU COULD DEVELOP TREATMENTS FOR DISORDERS. >> THAT'S GENERALIZED YES. WE START WITH FLIES. THEN WE HAVE TO SHOW THAT THE GENE THAT WE FIND HAVE HOMOLOGOUS FUNCTIONS IN HUMANS. AND THEN WE HAVE -- YES POTENTIALLY FOR SLEEP DISORDERS, THAT'S RIGHT. I JUST WANTED TO SAY ONE OTHER THING ABOUT THIS ACTIVITY ASSAY. I MENTIONED BEFORE THAT FIVE MINUTES WITHOUT AN ACTIVITY COUNT WAS DEFINED AS SLEEP. WHAT PEOPLE DID TO SHOW THAT WAS THEY WAITED UNTIL THEY HAD RESTED FOR ONE MINUTE AND THEN STIMULATED THEM. AND THEY RESPOND. THEY HAD THE SAME -- TWO MINUTES, THREE MINUTES FOUR MINUTES. BY FIVE MINUTES THEY WEREN'T RESPONDING SO THEY HAD THE INCREASED THE THRESHOLD. YOU HAD TO KNOCK ON THE TUBE TO GET THEM TO ARISE. SO THAT WAS ONE OF THE WAYS THAT WE SHOW THAT WE'RE NOT JUST SITTING QUIESCENT. NOT SITTING QUIETLY AS IT WERE, THEY'RE ACTUALLY SLEEPING. >> HOW DO YOU VISUALIZE STUDIES IN DROSOPHILA IMPACT THE QUESTION OF WHY DO WE HAVE SLEEP. WHY SLEEP. DO YOU HAVE SOME IDEAS AS TO HOW STUDIES IN FLIES IMPACT ON THE QUESTION OF WHY DO ALL SPECIES SLEEP. >> WHAT WE HAVE FOUND SO FAR ONE VERY INTERESTING THING IS WE FOUND A LOT OF GENE THAT HAVE TO BE -- TURNOVER WHICH I THINK IS REMARKABLE THAT SUPPORTS THIS IDEA THAT THERE'S SOME CHANGES IN PROTEIN SYNTHESIS THAT ARE -- DURING SLEEP AND WAKE. ANOTHER IS DIFFERENT LEVELS OF NEUROTRANSMITTERS ALTHOUGH WE'RE NOT MEASURING THEIR LEVELS, THIS GENOME WIDE ASSOCIATION IS DIFFERENT SNPs TRANSMITTERS WITH THE -- SEROTONIN FOR EXAMPLE. ANOTHER CLASS OF GENES THAT CAME UP THAT HAS TO DO WITH CENTRAL NERVOUS SYSTEM DEVELOPMENT. AND THAT BEGS THE QUESTION DOES YOUR BRAIN DETERMINE HOW MUCH YOU SLEEP, DO YOU NEED IT BASED ON THAT OR IS IT SOMETHING THAT THESE GENES ARE DOING IT AT THE ADULT PHASE. WE DON'T HAVE THE ANSWER TO HAT QUESTION YET BUT I THINK IT'S REALLY INTRIGUING. >> OKAY. WELL LISTEN, THANK YOU BOTH VERY MUCH.