>> MASUR AUDITORIUM AND THOSE WATCHING BY VIDEOCAST FOR OUR MAJOR LECTURE OF THE WEEK. WE'RE FORTUNATE TO HAVE DR. BRIGITTE KIEFFER WITH HER TRAINING IN IN BIOCHEMISTRY IN STRASBURG, FOR INSTANCE AND POST-DOCTORAL TRAINING IN BASEL BEFORE RETURNING TO STRASBURG. IN 2014 DR. KIEFFER MOVED TO McGILL UNIVERSITY IN MONTREAL WHERE SHE IS CURRENTLY A PROFESSOR IN THE DEPARTMENT OF PSYCHIATRY WHILE CONTINUING TO HOLD A PROFESSORSHIP AT THE UNIVERSITY IN STRASBERG IN FRANCE. SHE'S A GIANT IN THE STUDY OF THE OPIOID SYSTEM IN THE BRAIN. A TOPIC OF CONSIDERABLE CURRENT INTEREST AND IMPORTANCE GIVEN THE CRISIS WE FACE IN THIS COUNTRY. OF OPIOID OVERUSE AND ADDICTION AND OPIOID OVERUSE DEATHS. SHE'S THE FIRST PERSON TO ISOLATE A GENE ENCODING OPIOID RECEPTOR BACK IN THE EARLY 1990s AND THAT OPENED UP AN ENTIRE RESEARCH FIELD THAT LEADS TO MUCH OF OUR CURRENT UNDERSTAND OF THE MOLECULAR BASIS OF HOW OPIOIDS WORK AND WHAT THE ENDOGENOUS PURPOSE OF THAT RECEPTOR IS AND HOW MEDICATIONS AFFECT IT IN WAYS THAT CURRENTLY WE'RE ALL TRYING TO UNDERSTAND AND IN MANY INSTANCES TRYING TO MANAGE. HER GENETIC DIE -- DISSECTION HAS LED TO UNDERSTANDING PAIN DISORDERS AND CONTRIBUTED TO PHARMACOLOGY AND G PROTEIN COUPLE RECEPTOR RESEARCH SINCE THE OPIOID RECEPTOR IS IN THAT FAMILY. SHE'S BEEN WIDELY COLLABORATIVE AND SHARED MUCH OF HER INFORMATION AND GENETIC TOOLS AND CREATED RESEARCH LINES THAT HAD IMPORTANT IMPACT IN NEUROSCIENCE. WE'RE PROUD TO SAY SHE SAY GRANTEE OF THE NATIONAL INSTITUTES OF HEALTH DESPITE HER CURRENT LOCATION ABOVE THE BORDER AND RECEIVED NUMEROUS AWARDS FROM THE FRENCH AND U.S. ACADEMIES OF SCIENCE. THE FRENCH ACADEMY OF SCIENCE ELECTION IN 2013 AND IN 2014 RECEIVED AN AWARD FOR WOMEN IN SCIENCE. SO HER WORK HAS BEEN TRULY TRANSFORMATIVE FOR UNDERSTANDING THE OPIOID SYSTEM BUT WE HAVE LOTS OF WORK TO DO HERE. WE AT NIH ARE FOUND OF SAYING WE HAVE ALL HANDS ON DECK TRYING TO BRING THE BEST SCIENCE TO THE PROBLM WE CURRENTLY FACE. WITH OPIOID ADDICTION AND SHE WORK SHE'LL TELL YOU ABOUT TODAY WILL BE A CRITICAL PART OF ADDRESSING THAT. HER TITLE IS OPIOID RECEPTORS AND BRAIN FUNCTIONING SO JOIN ME IN WELCOMING DR. BRIGITTE KIEFFER. >> THANK YOU. VERY NICE INTRODUCTION. OKAY. GOOD AFTERNOON, EVERYONE. GOOD AFTERNOON TO YOU AND THANKS FOR COMING AND FOR ALL THOSE ON THE WEB. I AM VERY HAPPY AND HONORED TO GIVE THIS LECTURE THE WALS LECTURE AND HAPPY ALSO BECAUSE I MEET MANY COLLEAGUES HERE AT NIH, LONGSTANDING COLLEAGUES. I WANT TO TALK ABOUT THE OPITATE SYSTEM. I HAVE SLIDES. YOU DON'T HAVE THEM. CAN YOU TURN ON THE LIGHTS, PLEASE, THANK YOU. SO THIS IS A KIND OF TIMELY TOPIC AS YOU SAID. OPIUM IS A KIND OF MAGICAL SUBSTANCE THAT SEEMS TO RELIEVE ANY KIND OF PAIN. PHYSICAL PAIN, MENTAL PAIN, SOCIAL PAIN AND PRODUCES EUPHORIA. AND SINCE THE USAGE OF OPIUM WHICH GOES BACK TO THOUSANDS OF YEARS THE ACTIVE INGREDIENT WAS ISOLATED 200 YEARS AGO CALLED MORPHINE BY A GERMAN CHEMIST AND FROM THE ISOLATION OF MORE TEEN TEEN -- MORPHINE WAS USED TO TREAT SEVERE PAIN AND IT WAS OBVIOUS IT PRODUCED HIGHLY EFFICIENT ANALGESIA AND A STRONG PAINKILLER AND STILL THE STRONGEST PAINKILLER TODAY. AND HAD STRONG ACTIVE LIABILITY. SO SINCE THAT TIME THE FIELD IS AS A PAINKILLER AS MORPHINE BUT VOID OF ADDICTIVE LIABILITY. SO THE PROBLEM SEEMED TO BE SOLVES IN 1898 WHERE A GERMAN COMPANY A CHEMIST IN BAY ERN DECIDED TO GET A CHEMICAL REACTION. COMMERCIALIZED AS THE FIRST NON-ADDICTIVE COMPOUND AND IT WAS CALLED HEROIN. YOU KNOW TODAY IT'S MORE ADDICTIVE THAN MORPHINE BECAUSE IT CROSS THE BLOOD BRAIN BARRIER FASTER AND SINCE THAT TIME HEROIN BECAME A PROBLEM. IN FACT, OPIOID WHEN OPIUM AND OPIATE DRUGS HAVE BEEN A CHALLENGE SOCIETAL CHALLENGE SINCE MANY MANY YEARS. FROM THE OPIUM WARS YOU MAY KNOW ABOUT WHERE BRITISH TRADING POLICIES DEVASTATED CHINESE ECONOMY LEADING TO MILLIONS OF OPIOID-ADDICTED INDIVIDUAL TO THE PROHIBITION IN THE 1990 TO NOW WE ENTERED THE ERA OF THE OPIATE CRISIS WHERE THE OVER PRESCRIPTION OF OPIATES TO TREAT PAIN HAS ACTUALLY LED TO GET OUT OF CONTROL LEADING MANY PEOPLE TO BECOME ADDICTED TO OPIOIDS AND TO TRANSITION TO HEROIN OR FENTANYL USE AND INCREASED OVERDOSE AND I'M SURE MANY HAVE SEEN THIS. THAT'S AN ACUTE PROBLEM HERE. NOW, DURING ALL THESE YEARS, NEUROSCIENCE HAS DRAMATICALLY EVOLVED. WE HAVE DISCOVERED A FASCINATING SYSTEM THAT ACTUALLY I WILL TALK ABOUT TODAY. THE OPIOID SYSTEM. SO MORPHINE ACTS BY BINDING ON THE BRAIN ON RECEPTOR SITES. THE FIRST RECEPTOR SITES WERE DISCOVERED IN 1973 AND THEY WERE CALLED MU RECEPTOR SITES. THESE WERE BINDING SITES. THEN BECAUSE THEY WERE BINDING SITES IN THE BRAIN AND THIS WAS THE BEGINNING OF PHARMACOLOGY, IT BECAME OBVIOUS WE WERE NOT BORN TO LIVE IN POPPY FIELDS AND MAYBE THESE RECEPTOR SITES HAD A ROLE IN OUR PHYSIOLOGY AND THE FIRST PEPTIDE THAT ACTIVATE THESE OR BIND TO THESE RECEPTORS WERE ISOLATED TWO YEARS LATER AND THEY BELONG TO A FAMILY OF OPIOID PEPTIDES THAT DERIVED FROM FREE GENES YOU MAY HAVE HEARD ABOUT. THIGHS GENE CLEAVE TO SMALLER PEPTIDED. I SHOW YOU THREE MAJOR PEPTIDES. THESE ALL SHARE A COMMON SEQUENCE THAT INTERACT IN THE RECEPTOR SITE. AND IDENTIFYING RECEPTOR GENES IS MORE DIFFICULT SO IT TOOK 20 YEARS TO THE BINDING SITE AND THEIZELATION OF GENE AND -- ISOLATION OF GENE AND DONE BY EXPRESSION CLONING AND IN UCLA WE PUBLISHED THE SAME WEEK WHICH IS FORTUNATE SO WE GOT EQUAL RECOGNITION AND SINCE WE COLLABORATED WHICH IS GREAT. AND SOON AFTER THE CLOTH OF THE RECEPT -- CLONING OF THE GENE NOW WE HAVE THREE GENES ENCODING THE RECEPTORS AND IT TOOK ANOTHER 20 YEARS UNTIL WE COULD PRODUCE ENOUGH OF THE PROTEIN TO BE ABLE TO CRYSTALIZE THE RECEPTORS AND OBTAIN AND GET THE STRUCTURE BY ACCELERATED MICROSCOPY AND WE SAW THE RECEPTOR. I'M NOT GOING TO TALK ABOUT THE FOURTH RECEPTOR. THE RECEPTOR BOUND TO ANTAGONIST. WE HAVE THE ATOMIC STRUCTURE OF INACTIVE RECEPTORS AND YEARS LATER WE GOT THE FIRST STRUCTURE OF THE ACTIVE NEW OPIOID RECEPTOR. NOW WE KNOW A LOT ABOUT THIS PROTEIN. WE KNOW THEM ATOM BY ATOM. AND THESE RECEPTORS BELONG TO THE BIG G PROTEIN RECEPTOR FAMILY. WE HAVE THOUSANDS OF THE RECEPTORS. THEY'RE EXTREMELY IMPORTANT AS BIOMEDICAL TARGETS. HALF THE DRUGS ARE FOR USE TO TREAT PEOPLE AS ANTAGONIST OR MEMBERS OF THIS BIG FAMILY. IF YOU WANT TO THINK ABOUT THE OPIOID SYSTEM, WE CAN THINK OF THIS SYSTEM AS A SYSTEM THAT EXISTS IN OUR BRAIN TO TEACH US BENEFICIAL BEHAVIORS AND HELP US COPE WITH STRESS. SO THE SYSTEM IS IMPORTANT TO REGULATE REWARD AND AVERSION PROCESSES AND I'LL TALK A LOT ABOUT THAT IN THE UPCOMING TALK. IT'S A SYSTEM THAT'S EXTREMELY POTENT TO REDUCE PAIN AND COPE WITH STRESS. I WILL TALK A LITTLE BIT LESS ABOUT THAT AND ALSO REGULATES A NUMBER OF AUTONOMIC FUNCTION AND OTHER THINGS ON THE PERIPHERY. YOU SEE FROM THE SLIDE THE FREE RECEPTORS ARE DISTRIBUTED VERY DIFFERENTLY IN THE MOUSE BRAIN ON THE LEFT AND IN THE HUMAN BRAIN ON THE RIGHT. SO CLEARLY INDICATING THEY HAVE DIFFERENT FUNCTIONS. SO THIS LEADS ME TO THE OUTLINE OF THIS TALK. THE FIRST PART WILL BE FOCUSSED ON THE RECEPTOR BECAUSE THIS IS OUR PROBLEM IN THE OPIOID CRISIS. THE SECOND PART IS MORE GENERAL ABOUT THE ROLE OF THE FREE OPIOID RECEPTORS IN PHYSIOLOGY AND I WILL FOCUS ON MOOD STATE AND PATHOLOGY DRUG ABUSE. AND THEN ENTER SPECIFICALLY INTO BRAIN CIRCUIT TO SEE HOW THE RECEPTOR OPERATES WITHIN THE CIRCUIT AND THE FOURTH PART IS ABOUT A NEW APPROACH WE DEVELOPED IN OUR LAB WHICH I CALL TRANSLATION NEUROIMAGING WHERE WE TRY TO UNDERSTAND IN MICE HOW IT SHAPES BRAIN FUNCTION CONNECTIVITY. LETS START WITH THE FIRST PART. I TOLD YOU WE HAVE THREE RECEPTORS HIGHLY ANALOGOUS. WHICH RECEPTOR IS DOING THAT? IT'S WORK CREATING MICE LACKING A GENE. THE NEW OPIOID RECEPTOR GENE AND WE FOUND THAT ANY ACTIVITY OF MORPHINE WAS PURELY AND SIMPLY ABRATED IN THE ANIMALS. -- ABLATED IN THE ANIMAL AND WAS NOT PLEASURABLE. THERE WAS NO MORPHINE DEPEND EN AND NOTHING. AND WE INJECTED MORPHINE AND THE ANIMALS WOULD NOT DIE FROM OVERDOSE. IT WAS CLEAR FROM THE SIMPLE EXPERIMENT THIS RECEPTOR IS ACTUALLY THE SINGLE AND ESSENTIAL TARGETS THAT MEDIATES THE ADVERSE AND AFFECTS OF OPIOIDS WE USING IN THE CLINICAL OR ABUSED IN THE STREET. THIS IS THE ORIGINAL EXPERIMENT BUT THERE'S LATER THAT CON VIRTUAL THIS. WHAT YOU KNOW ABOUT OPIOID EFFECT IN THE CLINIC IN TERMS OF OPIOID USE DISORDERS ARE MEDIATED BY THIS PARTICULAR RECEPTOR. AND IF YOU USE MICE EITHER THE DEN TA OR CAPPA RECEPTOR THEY RESPOND PERFECTLY NORMALLY TO MORPHINE. PEOPLE THOUGHT IT WOULD HAVE BEEN NICE TO HAVE TWO RECEPTORS. ONE FOR PAIN OR NOCICEPTIVE CONTROL AND ONE FOR EUPHORIA AND WE WOULD DESIGN DRUGS THAT WOULD BE SPECIFIC TO ONE OF THE RECEPTORS. BUT WHAT IF NOW THERE'S A SINGLE RECEPTOR PROTEIN THAT MEDIATES ALL THIS AFFECT? HAPPILY, SCIENCE GOES ON AND THERE WERE A FEW REVOLUTIONS IN THE G PROTEIN RECEPTORS AND ONE REVOLUTION IS IN SIGNAL. LET ME GO THROUGH WHAT HAPPENS WHEN AN OPIOID RECEPTOR SIGNALS. THE AGONIST IT BINDS TO THE RECEPTOR WHICH IS THE BRAIN RECEPTOR, THIS WILL RECRUIT A NUMBER OF EFFECTERS SOME ON G PROTEINS AND IN THIS CASE THEY'RE INHIBITORY GENE PROTEINS WHICH RECRUIT OTHER EFFECTERS OR THERE MAY BE A RECRUITMENT OF NORMAL PROTEINS SO TWO MAIN PATHWAYS. CLEARLY THERE'S A WHOLE COMPLEX RESPONSIBLE TO INHIBIT THE ACTIVITY. THESE EFFECTERS CAN BE CHANNELS CMP, EVERY EVENT OF ACTIVATING AN OPIOID RECEPTOR IN THE NEURON AND THEN THERE'S A CASCADE THAT ARE ENGAGED AND GO TO THE NUCLEUS AND MODIFY TRANSCRIPTION. WE HAVE A GI COUPLED RECEPTOR. SO WHAT IS IMPORTANT IS NOT ONLY THE RECEPTOR, IT'S THE AGONIST RECEPTOR/EFFECTER COMPLEX. THIS IS AN EXAMPLE OF A SEARCH IN THE LITERATURE OF SIGNALLING EFFECTERS THAT HAVE BEEN IDENTIFIED AS THE EFFECTERS OF THE NEW OPIOID RECEPTOR. IT'S ABLE TO ACT ON ALL THE PATHWAYS. SO THIS LEADS US TO WHAT WE CALL BIASSED SIGNALLING OF FUNCTIONAL CONNECTIVITY. IT'S BEEN DEVELOPED AND IT'S NOT UNIQUE TO OPIOID RECEPTORS. IT'S GENERAL FROM THE RECEPTORS. THE CONCEPT SAYS THAT DEPENDING ON THE DRUG, THE ACTIVE CONFIRMATION OF THE RECEPTOR WILL DIFFER AND DIFFERENT SIGNALLING EFFECTERS WILL BE ENGAGED. THIS IS JUST A THEORETICAL EXAMPLE WHERE DRUG ONE WOULD BE WITH A AND B AND EFFICACY IS LOW AND DRUG TWO WOULD HAVE OPT MAT AND DRUG THREE WOULD BE BAD BECAUSE IT WOULD MAKE AN ADVERSE EFFECT. IT LED PEOPLE TO THINK ABOUT TRYING TO DESIGN A DRUG BY TARGETING SPECIFIC SIGNALLING PATHWAY AND SEPARATE GOOD AND BAD EFFECTS. SO REGARDING THE OPIOID RECEPTORS THERE'S A NICE RECEPTOR HERE. YOU SEE THE TWO PATHWAYS AND THE GIGO AND BETA ARRESTIN AND IF YOU DELETE IT IN THE MOUSE, MORPHINE HAS A BETTER EFFECT. NICE ANALGESIA AND LESS TOLERANCE SO IT SEEMED IF YOU AVOID THE BETA RESTING PATHWAY YOU MAY BE ABLE TO HAVE A BETTER DRUG. SO THIS IS HOW THE COMPANY DEVELOPED THE DRUG FOR CLASSICAL STRAINING EFFORTS BIASSED AND BINDS SPECIFICALLY TO THE GIGO AND DOESN'T HAVE BETA RESTIN AND IS NOW ENTERING CLINICAL TRIAL PHASE 3. THERE'S ANOTHER DRUG DEVELOPED AT SCRIPPS HIGHLY GI BIASSED. IT'S GOT TRAINING AND CHEMISTRY EFFORT AND I'D LIKE TO MENTION THE APPROACH USED. IT'S VERY IMPORTNT BECAUSE IT'S A FIRST CASE WHERE VIRTUAL DOCKING WAS PERFORMED. THIS IS EXTREMELY EFFICIENT BECAUSE NOW YOU CAN DOCK MILLIONS OF MOLECULES ON THE ATOMIC STRUCTURE OF THE RECEPTOR AND SELECT THOUSANDS OF COMPOUNDS AND RANK THEIR ABILITY TO BIND TO THE RECEPTOR AND SPECIFICALLY PICK CHEMICAL STRUCTURES THAT ARE TOTALLY NEW. THIS WAY, THIS GROUP WERE ABLE TO ISOLATE CHEMICAL STRUCTURES THAT WERE POTENTIALLY GOOD AND NOW YOU GO TO THE LAB AND SYNTHESIZE THE FIRST 20 AND WORK WITH THEM AND DEVELOPED THE MOLECULE PCDM21 GI BIASSED AND DOES HE -- DEPRESSION SEEMS TO BE LOWER. WHY IS THIS IMPORTANT? THIS APPROACH ACCELERATES THE PACE OF DISCOVERY. THIS IS WHERE THE RECEPTORS ARE IMPORTANT. WE NOW HAVE DRUGS THAT HAVE BINDING TO THE NEW NEUROSCIENCE AND I DON'T KNOW IF YOU CALL THEM OPIOIDS AND THERE'S A HOTLY DEBATED ISSUE IN THE FIELD. IS THE FACT THE COMPOUND IN THE CELL SYSTEM SHOW BIASSED ACTIVITY WHICH MEANS THEY DON'T ACTIVATE ALL THE EFFECTERS BUT PART OF THEM. IS THIS RESPONSIBLE FOR THE DIFFERENTIAL EFFECTS IN VIVO. THERE'S A BIG GAP BETWEEN THE TWO. WE'RE NOT SURE. AT THE MOMENT CORRELATION IS MADE AND WE HOPE IN THE FUTURE TO ADDRESS THIS BY LOOKING AT THE BRAIN CONNECTIVITY. I'LL GO BACK TO THIS IN MY LAST THIS LAST SLIDE IS TO TELL YOU THERE'S DIFFERENT WAYS TO MODULATE THE NEW OPIOID RECEPTOR SIGNALLING AND TRY TO DEVELOP NOVEL DRUGS BEYOND BIASSED AGONIST. ONE IS TO DEVELOPED THE REGULATOR YOU SEE ON THE LEFT SO THEY BASICALLY FACILITATE ANTIGEN SIGNALLING AND HAVE MODULATORS THAT DON'T WORK IN VIVO AS YET SO TO BE DEVELOPED. THERE'S ANOTHER WAY WHICH IS TO INCREASE THE LEVEL OF ANDOGEN PEPTIDE AND THIS SAY BLOCKER AND ANOTHER WAY TO INCREASE NEURORECEPTOR ACTIVITY BUT RIGHT NOW THERE'S NOTHING REALLY ON THE MARKET OR AN EFFORT TO DEVELOP WHAT DOESN'T ENTER THE BRAIN AND TO DEVELOP THE LIGAND AND USE OPIOID RECEPTORS TO MODULATE THE ACTIVITY. THE TAKE-HOME MESSAGE IS THERE'S MANY STRATEGIES UNDERWAY TO TRY TO REDUCE THE ADVERSE EFFECT MITIGATED BY ACTIVATED NEURORECEPTORS. I'LL STOP WITH THIS PART AND MOVE TO THE SECOND PART. >> WHEN YOU USE NEW RECEPTOR KNOCK-OUT MICE. THEY HAVE NO NEW RECEPTORS, I TOLD YOU THEY DON'T RESPONSE TO MORPHINE. SO MORPHINE IS NOT PLEASURABLE, FINE. YOU WILL SEE THE MICE ARE NOT INTERESTING OR THE DRUGS ARE NOT RE-ENFORCING WHEN THE NEW RECEPTOR IS GONE. IT TELLS US NEW RECEPTORS ARE NOT ONLY MITIGATING REWARDING ORDER S OF THE OPIATE THAT BINDS TO IT BUT REWARDING ORDER S OF OTHER DRUGS OF ABUSE LIKELY THROUGH THE ANDOGEN RELEASE. WE HAVE A MEDIATOR OF DRUG REWARD AND IF WE PUSH FURTHER WE CAN LOOK AT NATURAL REWARD. WHAT IS NATURAL REWARD? IT'S STIMULI OR SITUATION IMPORTANT TO LEARN BUT WE NEED TO DO THIS FOR OUR SPECIES AND SURVIVAL LIKE EATING IS A NATURAL REWARD. SEXUAL ACTIVITY, IS A NATURAL REWARD. SOCIAL INTERACTION IS EXTREMELY REWARDING AND SOCIAL REJECTION IS EXTREMELY PAINFUL. AND I'D LIKE TO MENTION WE TEST THE POSSIBILITY THAT SOCIAL INTERACTIONS ARE MEDIATED -- SOCIAL REWARDS, SORRY, REWARDED PARTLY THROUGH THE OPIOID RECEPTOR AND IT WAS DEMONSTRATED BY A PAPER WE PUBLISHED YEARS AGO THAT SHOWS RECEPTOR PUPS FOUR DAYS OLD SHOW REDUCED MATERNAL ATTACHMENT. TO EXPLAIN SIMPLY, MATERNAL BEHAVIOR IS REWARDING FOR THE PUP AND THIS IS IMPORTANT FOR THE PUP SURVIVE. THEY REACT TO THIS TO HAVE THEM DOING THIS AGAIN AND AGAIN. THIS IS HOW BONDING TAKE PLACE. THESE DO NOT EXPERIENCE THE PLEASURABLE FEELING OF MATERNAL BEHAVIOR AS A CONSEQUENCE, BONDING DOESN'T TAKE PLACE AS AS WELL. THIS HAS QUENCES AND WE DEMONSTRATED THAT MICE SHOW AUTISTIC LIKE BEHAVIOR. THERE'S AN IMPORTANT ROLE OF THE OPIOID RECEPTOR BEYOND DRUG ABUSE WHICH HAS TO DO WITH NATU NATURAL REWARD. IT'S VERY IMPORTANT. BRIEFLY, IF WE DO ALL THESE EXPERIMENTS USING THE DRUG RECEPTOR KNOCK-OUT MICE WE FIND THESE MICE SHOW HIGH LEVELS OF ANXIETY AND DISPARATE BEHAVIOR WE DON'T SEE IN THE CAPPA RECEPTORS. WE FOUND AT THE TIME IN 2000 THAT IN FACT THESE RECEPTORS HAVE ANTI-DEPRESSANT ACTIVITY AND ANXIETY AND LED DRUG COMPANIES OR CHEMISTS TO MOVE THE DRUGS DEVELOPED TO TREAT CHRONIC PAIN INTO PSYCHIATRY AND WERE SUCCESSFUL UP TO NOW TO DEVELOP THE AGONIST AS AN ANTI-DEPRESSANT. I BASICALLY SUMMARIZED STUDIES MADE BY MANY LABS. WE KNOW ACTIVATING THE NEURORECEPTOR IS PLEASURABLE AND IT'S CLEARLY IMPORTANT IN THE INITIATION OF ADDICTION. I SHOW THIS TO YOU IN THE NEXT SLIDE. WE KNOW OTHER THE OTHER HAND, I DIDN'T TALK ABOUT CAPPA. ACTIVATING THE CAPPA RECEPTOR IS HIGHLY DYSPHORIC AND THEY OPPOSE MODULATION OF THE TRANSMISSION WE ALSO KNOW IF YOU SWITCH TO A MOOD STATE REPEATED STRESS OR REPEATED EXPOSURE TO DRUG INCREASES CAPPA RECEPTOR AFFINITY AND THIS CONTRIBUTE TO THE NEGATIVE EFFECT OF SAY STRESS AND ADDICTED INDIVIDUALS. ON THE OTHER HAND, WE KNOW ACTIVATING RECEPTORS AND ANTI-DEPRESSANTS SO IT'S RECOGNIZED BLOCKING THE CAPPA IS A STRATEGY TO HELP TO IMPROVE MOOD. SO THIS IS PHYSIOLOGY. LET'S NOW GO TO PATHOLOGY AND THE ADDICTION CYCLE. APOLOGIZE TO THOSE WHO KNOW THES BY HEART BUT ADDICTION IS A COMPLEX BRAIN DISORDER AND DEVELOPS FROM CASUAL USE TO BINGE USE AND IT'S FOLLOWED BY WITHDRAWAL STATE AND THIS IS AN AB VERBIVE STATE AND FOLLOWED BY AN ANTICIPATION PERIOD KNOWN AS CRAVING AND THEN CRAVING LEADS YOU TO CONSUME AGAIN AND GO IN A KNEW BINGE INTOXICATION RESULT. AS YOU GO THROUGH THIS CYCLE, THE BRAIN ADAPTS AND THE DRUG IS LESS REWARDING BUT AVERSIVE EFFECT OF WITHDRAWAL ARE GETTING STRONGER. CRAVING IS GETTING STRONGER. MOTIVATION GETS BIASSED TOWARDS THE DRUG INSTEAD OF NATURAL REWARDS. SELF-CONTROL IS REDUCED AND ALL THIS LEADS TO REINFORCE THE ADDICTION CYCLE. WITHIN THIS PATHOLOGICAL PROCESS IT IS CLEAR NOW WE KNOW AND THE NEURORECEPTOR IS INVOLVED IN THE EARLY STEP OF DRUG CONSUMPTION. IT'S WELL ESTABLISHED THAT THE CAPPA RECEPTOR AND DIMORPHINE ARE INVOLVED IN THE MOOD OF WITHDRAWAL AND IT IS LIKELY THE RECEPTOR IS INVOLVED AND THERE'S EVIDENCE FOR THAT IN THAT PART. I DID PUT ON THE OTHER SIDE OF THE CYCLE, CAPPA BECAUSE THERE'S LITERATURE OF THE RECEPTOR ACTING IN HIGHER-ORDER PROCESSES BUT THERE'S MORE TO DO IN THIS ASPECT. I THINK THIS IS A FIELD OF RESEARCH WE'LL DEVELOP IN THE FUTURE. THE TAKE HOME FOUR IS THE RECEPTORS CONTRIBUTE VERY DIFFERENTLY TO ADDICTION. SO WHERE DO THE RECEPTORS ACTIVATE? UP TO THAT TIME WE TOOK THE RECEPTOR AWAY AND LOOKED AT BEHAVIOR AND IN THE MIDDLE IS A BIG BLACK BOX AND WE SAY IT'S IMPORTANT FOR THIS BEHAVIOR BUT WE CANNOT UNDERSTAND THE CHANGE IN BEHAVIOR IF YOU DON'T UNDERSTAND THE BRAIN'S CIRCUITS INVOLVED. THERE'S TWO WAYS TO DO THAT. YOU CAN MOVE IN AND THEN LOOK AT EVERY LITTLE TINY PATHWAY AND THESE ARE APPROACHES YOU CAN USE USING CONDITIONAL GENE KNOCK-OUT APPROACH, FOR EXAMPLE, AND OPTOGENETICS TO MANIPULATE THE NEURONS AND I'LL SHOW YOU A LITTLE BIT OF THAT. ANOTHER WAY IS TO ZOOM OUT AND LOOK AT THE WHOLE BRAIN AND SEE WHAT HAPPENS AND HOW THE NEURORECEPTOR ACTIVITY SHAPE THE BRAIN CONNECTOME. WE'RE DOING BOTH. I'LL SUMMARIZE IT IN THE NEXT SLIDE. BEFORE YOU DO A CONDITIONAL KNOCKOUT OF A RECEPTOR YOU KNOW YOU NEED TO USE MICE THAT EXPRESS IN SPECIFIC POPULATIONS AND WHERE YOU'LL TARGET THE KNOCKOUT. IN THE CASE OF OPIOID RECEPTOR IT'S A PROBLEM BECAUSE WE DON'T KNOW IN WHICH CELLS THE RECEPTORS ARE EXPRESSED. IT'S A GENERAL PROBLEM IN THE RECEPTOR FIELD AND ANTIBODIES IN VIVO ARE NOT GOOD AND IT'S DIFFICULT TO GET CELLULAR RESOLUTION. AND THIS IS AN OPPORTUNITY IN MY LAB DOING WELL NOW. HE'S BECOME TRULY AMERICAN. AT THAT TIME WE DEVELOPED THIS COLLABORATION AND WE DEVELOPED A KNOCK-IN MOUSE MODEL WHERE WE WOULD INTRODUCE A FLUORESCENT FRAME SO THE ANIMALS WOULD NATURAL LISH PRODUCE A -- NATURALLY PRODUCE A RECEPTOR THAT IS VISIBLE. WE HAD TO BE SURE THE RECEPT SER SER -- RECEPTOR IS FUNCTIONAL AND THIS IS THE FIRST ATTEMPT. THIS WORKED EXTREMELY WELL. SO WE CREATED THESE MICE THAT EXPRESS THE FUNCTIONAL RECEPTOR WHICH IS EXPRESSED AT PHYSIOLOGICAL LEVELS THROUGHOUT THE NERVOUS SYSTEM AND YOU CAN SEE THE RECEPTOR. IT WAS A MIRACLE AT THE TIME. IT TOOK SO MUCH TIME TO CLONE THE GENE AND YOU CAN SEE IT WAS A MIRACLE. THE CELL IS COMING FROM THE BRAIN OF THE MICE AND YOU CAN SEE THE RECEPTOR WHICH IS CONCENTRATED ON THE SURFACE AND NOW I THROW IN AN OPIOID AND YOU CAN SEE THE RECEPTORS IN REAL TIME CLUSTER AND THEN THESE GO I. SO THIS IS HAPPENING IN 20 MINUTES. I JUST SHOWED YOU THE CONTRACTING 8 MINUTES. AFTER THAT, YOU CAN TRY AND COME WITH AN AGONIST AND THERE IS NO RESPONSE. THE ANIMAL IS TOTALLY TOLERANT. THE RECEPTOR HAS DISAPPEARED. IT'S INSIDE THE CELL. IT'S NOT ACCESSIBLE ANYMORE. THAT IS AN INTERESTING PROCESS. JUST TO SUMMARIZE, MANY EXPERIMENTS WE DID THIS TOOL ALLOWED US TO REVISE THE RECEPTOR ANATOMY AND THERE'S MANY THINGS THAT WERE REVISITED ONCE WE COULD SEE THE RECEPTOR. SECOND, WE COULD SEE REAL-TIME TRAFFICKING WHICH ALLOWED US TO DIFFERENTIATE BETWEEN AGONISTS THAT WERE ABLE TO REDUCE ANXIETY BUT INTERNALIZING THE RECEPTORS, SOME WERE NOT AND TOLERANCE DEVELOPED DIFFERENTLY DEPENDING ON WHERE THE DRUG IS ABLE TO IN TERRIBLIZE T TERRIBLIZE -- INTERNALIZE THE RECEPTOR AND YOU CAN SEE THE INTERNALIZATION INTRODUCED BY THE PERIOD IS DIFFERENT FROM PHARMACOLOGIC INTERNALIZATION AND IN PHYSIOLOGY IT'S DIFFERENT. WE SHOWED THAT IN FACT IN THE PHYSIOLOGICAL SITUATION WHEN THE ANDOGEN PEPTIDE IT DISAPPEARED AND APPEARS IN A SMALL PROPORTION OF NEURON AND NEVER WIPE IT FROM THE SURFACE. THERE'S NO REAL TOLERANCE USING ENDOGENOUS PEPTIDE. WE CREATED NEW RECEPTORS AND WE HAVE NEW ONES THAT ARE RED AND YELLOW. WE HAD SOME THAT WERE FUNCTIONAL AND THIS ALLOWS US TO NOT KNOW WHICH CELLS THE RECEPTORS ARE EXPRESSED. TO MAKE A LONG STORY SHORT WITH THE KIND OF STUDIES WE'VE BEEN DOING. I SHOW YOU A SIMPLISTIC VIEW OF THE BRAIN AND SHOW YOU BRAIN REGIONS INVOLVED IN THE DIFFERENT STAGES OF THE ADDICTION CYCLE. SO BRAIN REGION ENGAGED IN THE BINGE INTOXICATION STAGE IN ORANGE AND BRAIN REGION INVOLVED IN THE AGGRESSIVE STATE OF WITHDRAWAL IN BLUE AND THE CIRCUITS INVOLVED IN THE CREATING ANTICIPATION STAGE. AND ALL THE REGIONS EXPRESS OPIOID RECEPTORS. SO WHEN THE BRAIN IS EXPOSED TO OPIOIDS YOU HAVE NEURONS FOR ALL THESE CIRCUITS. SO WHICH RECEPTOR IS THOUGHT TO BE RESPONSIBLE FOR PROCESSING? WELL, THE RESEARCH OVER 50 YEARS SHOWS NEURORECEPTORS IN THE VTA WHICH HOLDS THE NEURONS ARE ABLE TO DIFFERENTIATE DOPAMINE AND RELEASE IT AND IT CONTRIBUTES TO REWARD PROCESSING. NOW, IN TERMS OF GENETIC MANIPULATION. WE'VE BEEN ABLE WITH COLLEAGUES TO DELETE THE RECEPTOR IN THE DOPAMINE FIELD AND SEE WHAT THE RECEPTORS DO THERE AND TO DELETE THE RECEPTOR AND ARREST THE RECEPTORS IN THE NEURONS SO A SPECIFIC POPULATION WHERE RECEPTORS ARE MOSTLY EXPRESSED AND WE WERE ABLE TO SHOW THE RECEPTORS HERE ARE EXTREMELY IMPORTANT IN MOTIVATIONAL STATES. NOW, I LIKE TO SAY AT THIS STAGE THAT THERE'S OVERSIGHTS FOR OPIOID REWARD THAT HAVE NOT BEEN STUDIED THERE. YET. AND THESE MICE ARE BEAUTIFUL NEW RECEPTORS EXPRESSING NEURONS IN THE HYPOTHALAMUS AND THERE'S MANY PLACE WHERE'S IMMUNE RECEPTORS COULD PRODUCE OR FACILITATE REWARD PROCESSING. NOW, SO THE TAKE-HOME IS THAT FACILITATE DRUGS WITHIN REWARD CIRCUIT. THE RECEPTORS ARE ALSO EXPRESSED IN WHAT WE CALL AVERSION CENTERS. THE BRAIN CENTERS IN BLUE. THE QUESTION IS WHAT DO THEY DO THERE? WHAT HAPPENS IF THEY'RE EXCITED BY A DRUG OR OVERSTIMULATED BY CHRONIC EXPOSURE TO OPIATES. TO TELL YOU TWO STORIES. ONE IS THE STORY YOU MAY HAVE HEARD ABOUT THE TINY BRAIN STRUCTURES WHICH CONNECT FOUR BRAIN REGIONS AND IT'S CONSIDERED A BRAIN AVERSION CENTER THE AMYGDALA IS ACTIVATED IN DIFFERENT SITUATIONS. YOU SEE RECEPTORS IN THE AMYGDALA AND THIS IS THE AREA THAT HAS THE HIGHES NUMBER OF RECEPTORS. NO ONE STUDIED THEM HERE SO WE HAVE AN AVERSION CENTER AND WE WERE ABLE TO DELETE NEURORECEPTORS IN THE MEDIUM AMYGDALA AND THE RECEPTORS ARE THERE INTACT WHEREVER IN THE ANYWHERE IN THE BRAIN AND WHAT IS NORMALLY AVERSIVE IS MUCH LESS AVERSIVE. THE WAY WE INTERPRET THE RESULTS IS WE BELIEVE THE NEURORECEPTORS IN THIS AVERSION CENTER NORMALLY INHIBIT AVERSIVE ENCODING AND THIS VERY IMPORTANT. AND THIS IS A NEW ROLE OF THE NEURORECEPTOR WHICH COULD ACT AS AN AVERSION RECEPTOR AND CAN BE ANTI-AVERSION IN AVERSION CIRCUITRY. ANOTHER LAST THING I'D LIKE TO TALK ABOUT IS THAT WE HAVE BEEN DEVELOPING A MODEL WHAT WE CALL A MOUSE MODEL OF PROTRACTED ABSTINENCE TO OPIOID. YOU CAN EXPOSE MICE TO REPEATED OPIOIDS AND MAKE THEM HIGHLY PHYSICALLY DEPENDENT FOR A WEEK, COULD BE MORPHINE OR HEROIN AND THEN TAKE THEM THROUGH SPONTANEOUS WITHDRAWAL FOR AN A WEEK OR TWO WEEKS OR FOUR WEEKS AND YOU'LL SEE THE DEVELOPMENT OF DESPAIR LIKE BEHAVIOR AND SOCIAL WITHDRAWAL. SO WE WERE ABLE TO SHOW THAT THIS IS VERY LONG LASTING. IT WILL LAST FOUR WEEKS, 10 WEEKS FOR A MOUSE. IT'S VERY LONG. WE ACTUALLY DON'T KNOW HOW LONG. IT CAN BE PREVENTED BY AN UPTAKE INHIBITOR AND CAN BE BLOCKED WHICH IS BLOCKAGE OF THE CAPPA RECEPTOR. THE INTERESTING EXPERIMENT I WANT TO TELL YOU ABOUT IS WE DELETE THE NEURORECEPTOR IN THE NUCLEUS WHICH IS THE CENTER THE HOME FOR THE NEURON AND THEN SXOERD THE ANIMALS REPEATEDLY TO THE OPIOID AND LET THEM GO FOR FOUR WEEKS AND WHAT HAPPENS IS THAT THE SOCIAL WITHDRAWAL THAT WE SEE IN CONTROL ANIMALS DOES NOT DEVELOP IN THESE ANIMALS. WHILE OTHER SIGNS OF ABSTINENCE DEVELOP. SO THIS MEANS THE ACTIVATION OF THE NEURORECEPTOR IN THE CENTER OF THE BRAIN IS RESPONSIBLE IN THIS CASE FOR THE SOCIAL INTERACTION DEFICIT FOR THE EMOTIONAL DEFICIT WE FIND IN OPIOID ABSTINENCE. THIS IS TO SAY IT'S NOT ONLY A REWARD BUT WHEN YOU ACTIVATE NEW RECEPTORS YOU ACTIVATE AND MODIFY MANY CELLS AND NEURONS IN THE BRAIN AND THOSE THAT ARE IN THE AVERSION CENTER ARE AS IMPORTANT AND CONTRIBUTE TO THE PROBLEM. NOW I'LL MOVE TO THE LAST PART. CAN WE MOVE NOW FROM WHY WOND WE LOOK AT THE WHOLE BRAIN? AND MAYBE EVEN TRANSLATIONAL. THIS IS THE KIND OF PROJECT I ENTERED INTO ABOUT 10 YEARS AGO OR MAYBE LESS, SEVEN YEARS AGO. IN HUMAN RESEARCH, NEUROIMAGING IS INSTRUMENTAL AND ONE OF THE WAYS TO LOOK AT BRAIN FUNCTIONING. THERE'S A HOST OF LITERATURE ABOUT STUDIES THAT ARE ABLE TO LINK GENOTYPE TO PHENOTYPE TO SHOW BIOMARKER FOR DISEASE AND EVEN TO SHOW DRUG EFFECTS. IT'S A LITTLE BIT MORE DIFFICULT IN HUMANS. WHY WOULDN'T WE TRANSFER THIS TYPE OF SCIENCE INTO RODENTS? AND IN ADDITION, THE CHERRY ON THE CAKE, WE WOULD BE MECHANISTIC BECAUSE IN MICE, FOR EXAMPLE, YOU CAN DELETE ANY GENE YOU WANT, ANYWHERE YOU WANT AND CAN MODIFY OR ACTIVATE ANY NEURON POPULATION AND LOTS OF MANIPULATION TO UNDERSTAND WHAT HAPPENS AND LOOK AT THE CONSEQUENCES ON THE WHOLE BRAIN. WE DECIDED TO GO FOR GENE AND DRUG EFFECTS. SO WBFC MEANS WOLE BRAIN FUNCTIONAL CONNECTIVITY. HOW NETWORKS OF NEURONAL POPULATIONS TALK TO EACH OTHER. WE HAVE A LITTLE BIT OF A PROBLEM BECAUSE YOU SEE A MOUSE BRAIN AND A HUMAN BRAIN. THE MOUSE BRAIN IS VERY TINY. YOU NEED A VERY BIG MAGNET AND I I WAS WITH THE KING OF THE MRI EUROPE AND HAD A BIG GROUP AND THERE WERE SOME BRAVE PEOPLE WORKING IN ROW RODENTS AND CREATED WAYS TO LOOK AT THE MOUSE BRAIN AND ONE WAY WAS THE RESTING STATE SO YOU LOOK AT SLOW BLOOD LEVEL FUNCTIONATIONS IN THE BRAIN AND -- FLUCTUATIONS IN THE BRAIN AND RESTING STATE AND THE APPROACHES SHE USED WERE TRACKING AND I SHOW YOU THIS IS A STRUCTURAL MRI TO GIVE YOU AN IDEA OF THE TYPE OF RESOLUTION. HER RESOLUTION IS SIMILAR TO WHAT YOU CAN GET IN HUMANS. SO WE DECIDED TO TRY AND COMPARE CONTROL ANIMALS WITH KNOCK-OUT ANIMALS. SO BASICALLY A NORMAL ANIMAL WITH AN ANIMAL LACKING ONE GENE AND IS THERE ANYTHING WE CAN SEE AND COMPARING THE RESTING STATE. AND WE HAVE NO HYPOTHESES AND WE LOOKED AT THE BRAIN AND HAVE NO PRE-CONCEIVED HYPOTHESES AND SHE USED A COMPONENT ANALYSIS TO FIND ABOUT 100 COMPONENTS WHICH MEANS THEY FIT TOGETHER. THEY HAVE A SYNCHRONOUS ACTIVITY AND CORRESPOND TO ANATOMICAL BRAIN REGIONS WHICH IS GOOD. NOW WE CAN LOOK AT LONG-RANGE COMMUNICATION. WE LOOK AT SYNCHRONY AND HOW THE COMPONENTS FIT WITH TIME AND WHETHER WE HAVE SYNCHRONY OR NOT AND WE HAVE WILD TYPE AND KNOCKOUT AND HOW DOES EACH COMPONENT TALK TO THE COMPONENT AND OBSTRUCT THE MATRICES AND HAVE THE FINGERPRINT. THE FINGERPRINT SHOWS YOU THE DIFFERENCES BETWEEN THE WILD TYPE AND KNOCK-OUT ANIMAL. THE FACT THE OPIOID RECEPTOR WAS DELETED AND RESHAPES THE CONNECTOME AND IF YOU USE STATISTICAL ANALYSIS YOU'LL SEE MOST CHANGES OCCUR IN BRAIN AREAS THAT PROCESS NEGATIVE BALANCE INFORMATION. SO NEGATIVE EFFECT. YOU HAVE PAIN REGION, YOU HAVE THE AMYGDALA AND RING -- CINGULE T AND WE MADE A VOLUMETRIC ANALYSIS AND FOUND CHANGES IN THE KNOCKOUT THAT CORRESPOND TO THE FUNCTIONAL CHANGES THAT MADE SENSE AND WE CAN SEE FROM THE PREFRONTAL CORTEX AND HOW IT TALKS TO THE REST OF THE BRAIN AND WHAT WE CAN SEE IS THE MAJOR CHANGES IN THE KNOCKOUT MICE IS THE DECREASED DIALING BETWEEN THE NUCLEUS CIRCUMENS AND WE CAN USE A KNOCKOUT ANIMAL FOR ANOTHER RECEPTOR FOR A DIFFERENT FUNCTION AND WE FIND A VERY DIFFERENT FINGERPRINT IN THIS CASE, MAJOR CHANGES ARE IN THE CORTEX THAT FIT WELL WITH THE ANIMALS WITH AN HGD PHENOTYPE AND THE TAKE HOME IS THE MAPPING IS AVAILABLE IN THE MOUSE AND WITH WE'RE GOING APPLY THE TECHNOLOGY TO ABSTINENT ANIMALS OR ANIMALS EXPOSED TO OPIOIDS BECOME ABSTINENT AND TRY TO TREAT THESE ANIMALS AND SEE WHAT IS CHANGING IN THE BRAIN AND WE HAVE STUDIES WITH HUMANS AND HAVE COMPARED THEM WITH THE OPIOID RECEPTOR GENES AND WERE ABLE TO FIND IN POSTMORTEM TISSUE IT'S EXPRESSED IN THE HUMAN AMYGDALA AS COMPARED TO THE MOUSE AND WERE ABLE TO COMPARE DIFFERENT PHENOTYPE OF HUMAN INDIVIDUALS AND ABLE TO FIND THE AMYGDALA COMPONENT AND IF YOU COMPARE THE GROUP IT TALKS DIFFERENTLY TO OTHER PARTS OF THE BRAIN IN THE GG VARIANT INDIVIDUALS. SO THIS IS PROSPECTIVE AND WHAT WE'RE DEVELOPING. AND THE LAST PARTIED LIKE TO TALK ABOUT IS THE DRUG EFFECTS. WE'RE TRYING TO SEE NOW HOW NEW OPIOID AGONISTS MODIFY THE BRAIN CONNECTOME. WE STARTED A BIG STUDY WHERE WE COMPARE HOW MORPHINE, FENTANYL AND BUNEBUPONEFRINE AFFECT THE BRAIN. THEY'RE NOT ALL AS REWARDING WHEN YOU DO BEHAVIORS AND SOME HAVE BIASSED SIGNALLING PROPERTIES. THEY DON'T EFFECT SIGNALLING EFFECTERS THE SAME WAY. THE WAY WE DESIGNED THE EXPERIMENT IS TO TRY TO CAPTURE THE EFFECT OF AN ACUTE INJECTION. THE ANIMAL IS RECORDED IN THE RESTING STATE AND RECEIVE THE DRUG AND ACQUIRE IMAGES. WE DO THIS IF WE DO THIS IN KNOCKOUT MICE WE CAN TRACK EFFECTS AND LOOK AT RECEPTOR MEDIATOR EFFECTS. WE'RE ANALYZING THE DATA FOR NOW. WE CAN FOR EXAMPLE LOOK AT 14 DIFFERENT BRAIN REGIONS WHICH ARE INVOLVED AS SEEN WITH THE COLOR CODE CORRESPONDS TO THE DIFFERENT ASPECT OF OPIOID USE DISORDERS. AND EACH TIME AND NOW WE CAN COMPARE LOOK AT THE DRUG EFFECT. EACH TIME YOU SEE A LINE HERE IT TELLS YOU CONNECTIVELY BETWEEN THE TWO REGIONS IS DECREASED BY THE DRUG. NOTHING HAPPENS IN THE IS A LINE CONTROL AND MORPHINE CHANGES CONNECTIVITY DRAMATICALLY THE FIRST TIME PERIOD AND THEN GRADUALLY FADES AWAY AND BUNE BUE PREN ORE COMES UP LATER. WE CAN SEE HOW TO THE DRUG CHANGES WITH IT TALKS TO THE REST OF THE BRAIN AND FENTANYL AND BUPRENORPHINE HAS DIFFERENT EFFECTS BUT IF WE DO THE SAME THING IN THE KNOCKOUT MICE AND THEY LACK THE RECEPTOR, MORPHINE DOES NOT, FENTANYL DOES NOTHING SO THEY'RE SELECTIVE AND THE BUPRENORPHINE HAS ANOTHER EFFECT NOT MEDIATED BY THE NEW RECEPTOR AND THIS IS INTERESTING BECAUSE WE KNOW BUPRENORPHINE ALSO ACTS ON CAPPA RESENTERS. WE CAN THEN FOR EXAMPLE, FOCUS AND COMPARE AND SUPER IMPOSE DRUG EFFECTS ON THE SAME BRAIN AND YOU CAN SEE IMAGES LIKE THESE WHERE YOU CAN SEE, FOR EXAMPLE, MORPHINE, FENTANYL AND BURPRENORPHINE EFFECTS AND THE WAY THE AMYGDALA TALKS TO THE REST OF THE BRAIN. WE'RE AT THIS STAGE AT THE MOMENT. IT'S PRELIMINARY DATA AND LOOKING AT NEW APPROACHES TO CHARACTERIZE THE DATA AND CHARACTER IVESING -- CHARACTER CHARACTERIZING THE BRAIN STATE OF THE MICE TO BETTER UNDERSTAND THE EFFECTS OF THE BRAIN. WE HOPE TO BE ABLE TO PREDICT. WHAT IS GOING TO BE THE BEHAVIORAL EFFECT OF THE DRUG BY LOOKING AT FINGERPRINTS ON CONNECTIVITY AND HOPE IT WILL BE TRANSLATIONAL IN THE FUTURE. FOR NOW THERE'S ONLY ONE STUDY COMPARING BUPRENORPHINIC EFFECTS ON THE LAB RAT. I WILL CONCLUDE, THE FIRST PART TOLD YOU HA -- AND THIS TOLD YOU THIS THE NEURORECEPTOR IS THE GUILTY ONE LOOKING AT THE EFFECTS OF OPIOIDS AND MANY STRATEGIES ARE ON THE WAY TO REDUCE THESE ADVERSE EFFECTS AND THERE'S THINGS WE SHOULD DO. THERE SHOULD BE MANY MORE STUDIES FOR THESE DRUG DEVELOPMENT. THE SECOND POINT WAS THE RECEPTORS PLAY DIFFERENT ROLES IN HEDONIC STUDIES AND EMOTIONAL CONTROL AND I DIDN'T TALK ABOUT PAIN. IT'S A WHOLE FIELD. I THINK WHAT IS UNDERESTIMATED AND UNDER STUDIED ARE THE OPIOID PEPTIDES. WE KNOW MUCH LESS ABOUT THE PEPTIDES COMPARED TO THE RECEPTORS. THE FIRST PART WAS ABOUT THE OPIOID RECEPTOR IN ADDICTION. THINGS ARE MORE COMPLEX AND THEY DON'T ONLY FACILITATE REWARD, THEY DO OTHER THINGS IN OTHER PLACES OF THE BRAIN. IT'S GOING TO BE IMPORTANT TO BE ABLE TO TACKLE THE DIFFERENT NEURONS THAT RESPONSE TO THE OPIOIDS AND I INSISTED REVERSING THE NEGATIVE EFFECT OF PROTRACTED ABSTINENCE IS VERY IMPORTANT AND THE FOURTH PART IS METHODOLOGICAL AND WE CAN WORK IN THE MOUSE IN CONNECTING TO COULD BEECT OMS AND -- CONNECTOMES AND WE'RE AT THE CROSS ROADS OF DEVELOPING METHODOLOGICAL PROCESSES. I'D LIKE TO THANK MY TEAM AT McGILL AND INSIST ON THE FACT THAT NIH HAS BEEN FUNDING ME SINCE I WAS IN FRANCE AND MOVED TO CANADA. NIH HAS ALLOWED ME TO DO VERY RISKY PROJECT. I THINK THIS WAS VERY PRODUCTIVE AND DEEPLY THANKFUL TO OUR OTHER FUNDING SOURCES AND DEEPLY THANKFUL TO MY GUYS. THEY DID ALL THE WORK. I LOVE THEM. THANK YOU VERY MUCH. ONE I STUCK THERE BECAUSE HE WASN'T THERE THAT DAY. >> DR. COLLINS HAD TO LEAVE. HE HAD A PHONE CALL BUT WE WANT TO THANK YOU FOR THE ILLUMINATING TALK. WE HAVE ABOUT SIX MINUTES FOR QUESTIONS AND THERE'S A RECEPTION IN THE LIBRARY EVERYONE IS INVITED TO. YOU CAN COME AND TALK TO DR. KIEFFER AT THAT TIME BUT NOW QUESTIONS. >> THANK YOU. VERY THOUGHT-PROVOKING LECTURE. FROM THE OVERALL EFFECT OF THE TALK ON OPIOID WHAT I PICKED UP IS THAT IT PROBABLY HAS A DIVERSE AND MULTI EFFECT ON THE NATURAL EXISTING ANALGESIC AND ON OTHER RECEPTORS AND THE EFFECT AND CONSTIPATION AND THE MOLECULAR OR IMMUNE LOGICAL SIDE OF IT IS PROBABLY SUGGESTING THE ANALGESIC SIDE OF IT PROBABLY INDUCE THE GENES NOT JUST SPECIFIC FOR ONE RECEPTOR BUT EFFECT OTHERS. AND THERE'S DATA THAT IF IT DOES EFFECT THE DIABETIC PATIENT AND REDUCE COMPETITION AND INDUCTION OF I DON'T THE, ENHANCEMENT OF IN LINS RECEPTOR -- INSULIN RECEPTOR. THIS IS JUST A THOUGHT. >> WE KNOW THE RECEPTORS ARE ALSO EXPRESSED PERIPHERALLY. AND SYSTEM AND MAYBE ORGANS. THERE'S LESS STUDIED HERE BECAUSE THE EXPRESSION IS LOW BUT THERE COULD BE CONSEQUENCES PERIPHERALLY. >> IS THERE ANY TIME FRAME WHEN THE DEPRESSION OCCURRING THROUGH MAYBE MITOCHONDRIAL DYSFUNCTION? >> NO. THE NETWORKS IN THE BRAIN STEM RESPONSIBLE FOR THE DEPRESSION ARE WELL KNOWN. THE NUCLEI IN THE BRAIN STEM. THE RECEPTORS ARE IN DIFFERENT NUCLEI AND INHIBIT THE CELLS AND RESTORATION DROPS. AND THEY ORIGINATE FROM RECEPTORS OF THE SENSOR PHASE AND THERE'S RECEPTORS THERE AND ONCE THEY'RE ENDOCYTOSIS THEY KEEP SIGNALLING AND THERE'S INTERNAL SIGNALLING MECHANISMS. >> THANK YOU. >> IT COULD HAPPEN IN ANY NEURONS OR CELLS. THANK YOU. >> AT THE GENE LEVEL AT THE THREE RECEPTORS IS THERE VARIABILITY LIKE SINGLE NUCLEOTIDE OR POLYMORPHISMS THAT PUT AN INDIVIDUAL INTO THIS SUBGROUP OR THAT SUBGROUP. >> I HAVE SHOWN YOU ONE SNIP WELL KNOWN FOR THE NEW OPIOID RECEPTOR. THIS IS CALLED A1ATG. IT'S CHANGING THE INTERNAL SEQUENCE OF THE PROTEIN FROM AN AS POTTIC ACID AND MODIFIES THE RECEPTOR ON THE SURFACE AND PROBABLY MODIFIES FUNCTION. THIS IS OCCURRING AND PRETTY FREQUENT AND OCCURS IN 10% TO 15% OF THE POPULATION AND THE MOST STUDIED. THERE ARE OTHERS LESS WELL STUDIED BUT THEY ARE FOR THE THREE RECEPTOR GENES, YES. >> OKAY. THANK YOU. >> SO THERE'S A LOT OF MEDICINAL CHEMISTRY EFFORTS GOING ON WITH NEW AND KAPPA MOLECULES AND MANY STUDY AGONISTS LIKE THE NEW CAP AGONIST. I'M WONDERING WHAT DO YOU THINK THE PROSPECTS ARE FOR THINGS LIKE THAT IN TERMS OF SUCCESSFUL DRUGS AS WELL AS AND MAYBE YOU CAN INTERPRET THE ANSWER IN TERMS OF KNOCK-OUT ANIMALS. >> WELL, ANIMALS HAVE SPECIFIC PHARMACOLOGIC PROPERTIES AND WE DON'T HAVE PROOF THEY BIND SECTORS. WHY NOT? AND THIS QUESTION COMES BACK ALL THE TIME IT'S VERY DIFFICULT TO SHOW IN VIVO. 'M NOT SURE THIS HAPPENS REALLY. >> OKAY. >> SO WHY NOT, I WOULD SAY. >> THERE'S NEW RECEPTORS THAT CAN MEDIATE A REWARDING EFFECT AS WELL ADVERSIVE EFFECT. WHY DOES THE REWARDING EFFECT WIN IN THE BEGINNING. SO INITIALLY REWARDING SO BUT IT SHOULD BE STIMULATED. >> BUT IN FACT IT DEPENDS ON HOW THE RECEPTOR IS EXPRESSED. WHAT I SAID IN THE NAIVE ANIMAL, THE RECEPTOR THAT IS FOR EXAMPLE IN NEURONS WILL PROBABLY BE REWARDING. IN THE HABENULA THE RECEPTOR PROBABLY INHIBIT OR SHUTS DOWN. THEY DO THE SAME THING -- NO, IT GOES IN THE SAME DIRECTION, OKAY. NORMALLY YOU HAVE AN ANTI-AVERSIVE AND PRO-REWARD MU. WHAT HAPPENS WHEN THE ANIMAL IS EXPOSED TO OPIOID THE RECEPTORS ADAPT. WE KNOW THE REWARDING EFFECT OF DRUGS IS GETTING LOW PERP WE DON'T KNOW WHICH -- LOWER. WE DON'T KNOW WHICH RECEPTORS GET TOLERANT WHERE. WHAT IF THE RECEPTORS IN THE HABENULA ARE NOT ABLE TO INHIBIT THE AVERSIVE NEURONS AND HAVE YOU A NEGATIVE EFFECT THAT DEVELOPS AND WHAT WE'VE BEEN ABLE TO DO NOW WHERE WE HAVE NICE WITH RECOMBINASE WE DON'T KNOW AND IT'S WHAT WE HAVE TO DO NOW. >> WHY AREN'T DOPAMINE AGONIST DRUGS OF ABUSE? THEY'RE NOT AT ALL, RIGHT? WHY AREN'T PSYCHO STIMULANTS ABUSED? THEY'RE DRUGS OF ABUSE. >> PURE AGONISTS ARE NOT. >> I CAN'T ANSWER THAT QUESTION. I'M SURE THERE'S AN ANSWER TO THAT. I HAVE TO THINK ABOUT IT. >> OKAY. I WANT TO THANK YOU AGAIN. IT'S A BRILLIANT TALK AND COME TO THE RECEPTION AND TALK TO BRIGITTE ONE-ON-ONE.