DRUGS OF ABUSE. I THINK PROBABLY THE BEST WAY TO FIND OUT WHAT HE'S DOING IS TO HAVE HIM SPEAK TO YOU, TITLE TODAY IS "TARGETING RECEPTOR HETEROMERS FOR THE TREATMENT OF NEUROCRICK DISORDERS." >> THANK YOU VERY MUCH FOR THIS INVITATION. MY INTEREST FOR RECEPTOR HETEROMERS IS WITH MY INTEREST, MECHANISM OF ACTION RESPONSIBLE FOR THE FIX OF CAFFEINE. THE FAMOUS COCAINE AM FELT MEAN INCREASE PRODUCTIVITY ALLOWS ALSO HAS -- NEVERTHELESS CAFFEINE CAN CAUSE DEPENDENCE IT DOESN'T HAVE THE STRONG AFFECTS OF COCAINE AMPHETAMINE. IT'S VERY WELL ESTABLISHED THAT THE ACTIVATION AND EFFECTS OF STIMULANTS ARE DUE TO THE LIVIDITY TO INCREASE, TO STIMULATE THE SYSTEMS. THE ABILITY TO BIND TO THE DOPAMINE TRANSPORTER, IMAFEN MEET REVERSES IT WITH CONCOMITANT INCREASE IN ACTIVATION OF MULTIPLE SYNAPTIC RECEPTORS. WE HAVE NOW RECEPTORS. SINCE THE BEGINNING OF THE '90s WE KNOW THAT MAIN PHARMACOLOGICAL MECHANISM OF CAFFEINE IS THE COMPETITIVE ANTAGONISM. WE ALSO KNOW, WE KNEW THAT CAFFEINE IN CONCENTRATIONS REACHED IN THE BRAIN AFTER DRINKING COFFEE WOULD BE MAINLY RESPONSIBLE FOR THE EFFECTS WOULD BE THE BLOCK OF ENDOGENOUS ON A1 AND A2 RECEPTORS. A2 RECEPTORS ACTIVATED BY HIGH CONCENTRATIONS. HAVE NO AFFINITY FOR CAFFEINE. THE QUESTION WAS HOW MODULATING THE DOPAMINE SYSTEMS. THE FIRST CAME FROM SOME STRANGE UNEXPECTED RESULTS USING A MODEL. THE '23458 SYS OF THE BEHAVIOR IN RATS WITHIN NATURAL REGION OF THE PATHWAYS BY INJECTION. THEN WE STUDY BEHAVIOR INDUCED BY DIFFERENT DRUGS JUST BY LOOKING AT THE DIRECTION OF THE BEHAVIOR WE CAN TELL WE'RE DEALING WITH INDIRECT DOPAMINE AGONIST. IT'S LIKE AMPHETAMINE OR COCAINE. BECAUSE DOMINANT RELEASE OF DOPAMINE. THIS RESULT TRANSLATES TO SAWMILL AT THE TIME TRICK ACTIVITY WITH BEHAVIOR, IN THIS CASE. ON THE OTHER HAND LIKE D1 SELECTIVE AGONIST, SKS, 393 PRODUCE OPPOSITE EFFECT. CONTRA LATERAL AFFECT. IN THAT DUE TO OBVIOUSLY PREDOMINANT ACTIVATION OF THE DOPAMINE RECEPTORS. THEN IN THE INSTITUTE, IN BARCELONA IN THE UNIVERSITY WE FOUND OUT THAT CAFFEINE USING THIS MODEL WAS PRODUCING A CONTRA LATERAL TURNING BEHAVIOR. THAT WAS SURPRISING THAT WAS INDICATING THEY HAVE TO BE SOME KIND OF ACTIVATION OF DOPAMINE RECEPTORS, BUT WHAT KIND OF MECHANISM WOULD BE BEHIND THIS. THEN MY FIRST POST DOCTORAL TRAINING, IN THE INSTITUTE I HAD CHANCE TO WORK IN THE LABORATORIES AND WHAT WE DID WAS TO STUDY THE POSSIBILITY OF INTER-ACTIONS BETWEEN DOPAMINE RECEPTOR TO EXPLAIN THOSE RESULTS OF CAFFEINE. WE STARTED WITH THE VERY SIMPLE MODEL. WHICH IS RECEPTOR MODEL. PRODUCES STRONG MODEL. THIS IS COUNTER ACTED BY ADMINISTRATION OF DOPAMINE. FOR INSTANCE THE RESULTS WE HAD WITH THE D2 RECEPTOR. THEN WHAT WE FOUND IS THAT SEVERAL DIFFERENT AGONISTS WOULD PRODUCE DOSE DEPENDENT INHIBITION. THE DIFFERENCE, FOR INSTANCE, DIFFERENCE IN THE POTENT SEES WOULD TELL US THAT THE MAIN RECEPTOR INVOLVED. ALSO WHAT WE FOUND THAT IN CAFFEINE THEY WOULD PRODUCE THE OPPOSITE AFFECT AS EXPECTED. AFTER THESE RESULTS WE POSTULATED FOR THE FIRST TIME THE EXISTENCE OF POST SYNAPTIC INTEROCEPTORS. THE IDEA WOULD BE THAT THEN DOPAMINE D2 RECEPTOR ANTAGONIST WOULD WORK AS D2 AGONIST. WOULD BE THE SAME PHARMACOLOGICALLY THAN DOPAMINE D2 ANTAGONIST. IN FACT WE FOUND FOR THE FIRST TIME THAT THE CENTRAL ADMINISTRATION OF SELECTIVE A2 RECEPTOR AGONIST, PRODUCES DOSE DEPENDENT IN THE RAT. THEN IN THE LABORATORIES WE FOUND THE EVIDENCE FOR A MORE DIRECT INTER-ACTION BETWEEN THESE TWO RECEPTORS, A2A THAT WAS BY USING STRIATAL MEMBRANE PREPARATIONS. DOPAMINE VERSUS DOPAMINE D2 RECEPTOR. INCREASING CONCENTRATIONS OF DOPAMINE, THOSE DEPEND ON BINDING OF THE D2 RECEPTOR. WHAT HAPPENS THEN IN THE MEMBRANE PREPARATIONS, THERE IS SIGNIFICANT SHIFT TO THE RIGHT OF THIS CURVE. THERE'S DECREASE IN THE ABILITY OF DOPAMINE TO DISPLACE DOPAMINE D2 RECEPTOR ANTAGONIST. WHEN WE ANALYZE THE PARAMETERS OF THESE EXPERIMENTS WE FOUND THAT IS DUE TO DECREASE IN THE AFFINITY OF THE DOPAMINE D2 RECEPTORS FOR DOPAMINE. THESE RESULTS DEMONSTRATED THE KIND OF CLOSE INTER-A BETWEEN THE PREPARATIONS. BUT AT LEAST WHAT THEY DEMONSTRATED THAT THESE RECEPTORS ARE REALLY LOCALIZED IN THE SAME NEURAL ELEMENTS. THE INPUTS AND END PUTS IN THE TYPES MSN, WHICH CONNECT THE -- OUTPUT STRUCTURES OF THE BASAL GANGLIA. WE HAVE -- WHICH CONNECT WITH THE OUTPUT STRUCTURES, THESE NEURON CONTAINS PEPTIDES SUBSTANCE AND ALSO DOPAMINE RECEPTORS SELECTED WITH THE D1 THEN WE HAVE INDIRECT MSN WHICH CONNECTS THIS WITH THE -- WE FOUND FROM THE STUDIES OF THE GROUP THAT A2 RECEPTORS ARE SELECTIVELY LOCALIZED IN THE D2 RECEPTORS IN THE INDIRECT. BY THEN WE WERE STARTING RECEPTORS ARE LOCALIZED PARTICULARLY IN THE BRAIN, THE EXPRESSION MRNA AND LOCALIZED SICKLY IN THE DIFFERENT SPECIES. THE TECHNIQUE THAT WORK FOR FIRST TIME DOING EXPERIMENTS WITH THE APPROACH. WE WERE IN -- USE THAT PROBE TO PERFUSE DIFFERENT SUBSTANCE OF DOPAMINE RECEPTORS. THAT WOULD BE LOCALIZATION OF THE TERMINALS OF THE INDIRECT MSN WITH THE IDEA FUNCTIONALLY FUNCTIONALLY -- THE A2A AGONIST. THE RECEPTORS WOULD COUNTERACT THAT AFFECT. WHEN WE WOULD PERFUSE WITH THAT WE WOULD INCREASE THE AFFECT. THESE EXPERIMENTS WERE DEMONSTRATING A NON-CORRELATE IN THE INDIRECT MSN OF THOSE INTER-ACTIONS BETWEEN ANTAGONISTIC INTER-ACTIONS. LET'S START WITH RECEPTOR. THE RECEPTOR CONCEPT WAS INTRODUCED IN 1878 INDEPENDENTLY AND SIMULTANEOUSLY BY JOHN NEWPORT LANGLEY AND PAUL EHRLICH. WE KNOW NOW THAT THESE RECEPTORS PARTICULARLY THEY HAVE COMPLEXES WHICH INCLUDE AUTO RECEPTORS WITH THE POSSIBILITY TO FORM RECEPTOR HETEROMERS. AT LEAST TWO FUNCTIONAL RECEPTOR UNITS, WITH CHEMICAL PROPERTIES, THIS IS VERY IMPORTANT WITH CHEMICAL PROBLEMS THAT ARE DIFFERENT FROM THOSE OF COMPONENTS. WE CREATE NEW FUNCTIONAL ENTITY. THE DIFFERENCE OF BIOCHEMICAL PROPERTIES INCLUDE SPECIFIC LIGAND GENE PROTEIN AND ALSO SPECIFIC SIGNALLING. AND THIS BY CHEMICAL PROPERTIES MIGHT BE LIGAND INDEPENDENT, JUST BY FACT THAT THEY GET RECEPTORS, THEY ACQUIRE THE NEW PROPERTIES OR CAN BE DUE TO THE ACTIVATION OF LIGAND MEDIATED PROCESS. BECAUSE ONE OF THE -- IS BINDING A LIGAND WHICH I WANT TO REPRESENT. IN THIS CASE IT IS LIGAND THAT INDUCES CHANGES IN LIGAND RECOGNITION, FOR INSTANCE STIMULATION OF THIS RECEPTOR CHANGES THE BINDING CHARACTERISTICS. IT WILL HAVE SPECIAL NAME WHICH WE'LL CALLALOSTERIC INTER-ACTION. THIS IS HETEROMERS. THE DISCOVERY OF THE FIRST. THOSE WERE THE RECEPTOR HETEROMER. THE FIRST WAS THE GABA-B1. BUT WE CANNOT ACCORDING TO THE -- CANNOT INCLUDE HIM AS RECEPTOR BECAUSE THIS GAB BEEN B1 POET MER IS LEADED -- NEEDED FOR THE BINDING. YOU NEED TWO POET MERS TO HAVE FUNCTIONAL UNION OTHERWISE THEY'RE NOT FUNCTIONAL IF THEY'RE NOT -- PROBABLY FIRST RECEPTOR HETEROMER WE SHOULD SAY ONE DISCOVERED IS THE OPENNOID RECEPTOR HETEROMER THAT WAS IN 1999. THE NEXT YEAR WE CAME -- DISCOVERY WE AND ANOTHER GROUP RESTEP FOR INCLUDE -- A DOPAMINE AND DOPAMINE D2. SINCE THEN, SINCE 2005 HAS INCREASED VERY SIGNIFICANTLY. BUT LET'S TALL IT PUTATIVE, LIST OF POUTTIVE RECEPTORS STILL SOME OF THEM THEY DON'T COMPLY WITH THE CRITERIA FOR IDENTIFICATION OF RECEPTOR HETEROMERS, LIKE TRANSECTED CELLS OR EVEN UNLESS -- THIS CRITERIA IS PUBLISHED IN A MEETING OF LEADERS IN THE FIELD THAT WAS ORGANIZED BY NATIONAL INSTITUTES OF HEALTH, IN PARTICULAR BY NIDA. OF COURSE ONE IS D2 RECEPTOR. IN THIS STRAY @TIM IS THE BASAL ANGRY LA THIS IS PART OF BRAIN CIRCUITS INVOLVED IN THE LEARNING. THAT INCLUDES PARKINSON'S DISEASE, SCHIZOPHRENIA, OBSESSIVE COMPULSIVE DISORDER AND SO ON. WE GO BACK TO THE MSN, THIS CONSTITUTES MORE THAN 95% OF NEURAL POPULATION, TOO MANY INPUTS. THIS INPUTS CONVERGE IN THE DENDRITIC SPINES, HAVE MULTIPLE DENDTRITIC SPINES. IT MAKES CONTACT WITH THE HEAD OF THE DENDRITIC SPINE AND DOPE NEWER QUICK MAKES SIGH NAP PARTICULAR CONTACT WITH THE SPINE. THEN THESE EARL DENTS, TERMINAL AND PROCESSES THAT WRAP AROUND THE SYNAPSE CONSTITUTE WHAT WE CALL TRY A LITTLE SPINE MODULE. DEFINE AS LOCAL MODEL. WHICH WE -- WHICH WE HAVE DEFINED AS ONE OR MORE NEURONS OR ONE OR MORE GLIAL CELLS THAT OPERATE AS AN INDEPENDENT UNIT. WHY IS THIS SO IMPORTANT FOR ME. IT'S BECAUSE THIS CONCEPT THAT ALLOW US TO UNDERSTAND QUITE WELL WHAT IS THE MEANING, FUNCTIONAL MEANING OF EXTRA SYNAPTIC TRANSMISSION, IS IN NATURAL PARTICULAR RECEPTORS, AND ALSO RECEPTOR -- WE GIVE YOU EXAMPLE OF WHAT I MEAN. IN OUR TRY A LITTLE MODEL, DOPAMINE IS NOT ONLY RELEASED SYNAPTICALLY GOES IN PREAND POST SYNAPTIC ELSEMENTS. GLUTAMATE IS NOT ONLY RELEASING SYNAPTICALLY BUT STILLS OVER THE SIGH NAPS TO STIMULATE RECEPTORS BOTH AT THE DOPANERGIC SYNAPSE. THIS IS IMPORTANT FOR OUR -- THAT ALSO GLIAL PROCESSES OF THE MAIN SOURCE OF -- THAT COMES MOSTLY FROM ATP THAT IS RELEASED FROM THIS PROCESSES, CONVERTED THEN BINDS TO RECEPTORS THAT ARE LOCATED IN DIFFERENT ELEMENTS OF THE TRY A LITTLE SPINE MODEL. WE HAVE THE TRANSECTED CELLS. SECOND, IDENTIFY UNIQUE BIOCHEMICAL PROPERTY OF THESE RECEPTOR HETEROMER WHICH WE USE TO IDENTIFY. WE'LL SEE COUPLE OF EXAMPLES. AND THIRD WE FRY TO DEMONSTRATE THE FUNCTIONAL SIGNIFICANCE OF THE RECEPTOR IN THE BRAIN. WE REDEMONSTRATE IN OUR CASE WE USE ESTABLISHED METHODS -- YOU HAVE HERE IS MOLECULE, THE DONOR IS A MOLECULE THAT -- WE HAVE SUBSTRAIT AND THESE MOLECULE USUALLY REALLY LEVEL, EMISSION OF LIGHT MAKES TRANSFER OF ENERGY TO AN ADJACENT MOLECULE USUALLY GFB IN THIS CASE YFP WHICH ALSO TRANSMITS LIGHT BUT IN ANOTHER WAVE LENGTH. WHAT IS IMPORTANT ABOUT THIS PHENOMENON ONLY HAPPENS IF THE MOLECULE ARE AS CLOSE AS TEN NANOMETERS. APPLY THIS TO HETEROMERS. USUALLY IN THE C TERMINAL OF THE RECEPTORS. THE OTHER ONE, MOLECULE IN THIS CASE GFP TO THE OTHER RECEPTOR. THEN TRANSECT THEM TO MAMMALIAN CELLS THEN HAD SUBSTRAIT FOR VOW SIEVE RACE PROBABLY NOT IN CONTACT THEY ARE NOT CLOSE ENOUGH TO SEE THE BRAIN. TWO RECEPTORS REALLY IN CONTACT WE SEE EMISSION OF LIGHT FROM THE -- TRANSFER OF INFORMATION THAT MAKES GFB TO ELITE LIGHT AS WELL. POSITIVE VALUES IS NOT ENOUGH TO SAY THAT YOU HAVE RECEPTOR HETEROMERIZATION. IT COULD BE RANDOM CONDITION OF THE RECEPTORS IN THE TRANSECTED CELL. IN THAT CASE WE WOULD BE TALKING ABOUT NONSPECIFIC INTER-ACTION BETWEEN THE MOLECULES. IN ORDER TO DEMONSTRATE, WE SAY A SEPARATION OCCURS. THAT'S MADE OF SEVERAL EXPERIMENTS WHICH WE ARE TRANSECTING THE SAME AMOUNT OF THE RECEPTOR FUSED TO THE DONOR. AND INCREASING AMOUNTS OF THE RECEPTOR FUSED TO THE ACCEPT OR MOLECULE F. WE END UP WITH A LINEAR CURVE THAT MEANS IT'S NOT SPECIFIC. OTHERWISE WE SEE THIS KIND OF SEPARATION CURVE THAT WOULD INDICATE SPECIFIC THE HETEROMERIZATION. THAT'S QUITE ACCEPTED. WHAT WE NEED WAS TO FUSE TWO RECEPTORS WITH LUCIFERASE AND THE RECEPTOR TRANSECTING THE CELLS, FIND VERY NICE CURVE. WE ALSO FOUND IN OUR EXPERIMENTS TWO -- THAT WERE REALLY IMPORTANT IN THE INTER-ACTIONS IN THE HETEROMERIZATION. ONE OF THESE IS THE LOOK OF THE RECEPTOR THE OTHER CONTAINS PHOSPHORYL PLATED THAT IS 374. THESE WERE ESTABLISHED VERY STRONG ELECTROSTATIC INTER-ACTIONS. WHEN WE SUBSTITUTE THIS THEN THERE IS STRONG REDUCTION INSTEAD OF THE WILDTYPE. THEN WE SEE VERY STRONG REDUCTION BUT NOT IN THE APPEARANCE STILL HAS SEPARATION. THIS INDICATED THAT THESE ELECTROSTATIC INTER-ACTION NOT REALLY COMPLETELY NECESSARY BUT TO GIVE THE STRUCTURE OF THE HETEROMER. THEY HAVE CHANGED EITHER ORIENTATION OR THE DISTANCE BETWEEN THEM. IN ANY CASE, THAT WILL INDICATE THAT THESE TWO ARE INVOLVED IN METRO MER SAKES AFTER ALL. SECOND STEP WAS IDENTIFICATION OF THE BIOCHEMICAL PROPERTIES OF THE RECEPTOR HETEROMER. WE CAN USE THEM AS BIOCHEMICAL FINGERPRINT WITH A CHARACTERISTIC WHICH CAN BE USED FOR IDENTIFICATION IN AN ACTIVE TISSUE. OUR STUDY OF INTER-ACTIONS IS A COMMON PROPERTY OF RECEPTOR HETEROMER WE HAVE FOUND MANY EXAMPLES. IT'S INTER-MOLECULAR INTER-ACTION WHICH BINDING OF A LIGAND TO ONE RECEPTOR UNIT IN THE RECEPTOR HETEROMER CHANGES THE BINDING PROPERTIES OF ANOTHER RECEPTOR UNIT. THE BEST INTER-ACTION THAT WE DISCOVERED IN PREPARATIONS BY WHICH STIMULATION OF THE A2A RECEPTOR DECREASES AFFINITY OF DOPAMINE. WE HAVE EVIDENCE NOW THAT THESE EVENTS ON HETEROMERIZATION BECAUSE IN OUR CASE ALSO GROUP HAS DEMONSTRATED IF ELECTROSTATIC, WE DISRUPT THE INTER-ACTION. FOR EXAMPLE, IS RESULT IN PREPARATION, THIS IS D2 RECEPTOR AGONIST. BINDING OF THIS AGONIST IS DECREASED BY INCREASING CONCENTRATIONS. NOW IF WE USE WILDTYPE HAVE MUTATED CONTAINED, WHEN WE SEE IS VERY DECREASE POTENCY IN THE AFFECT OF SGS. THESE RESULTS DEMONSTRATE THAT THIS OWE ELECTROSTATIC INTER-ACTION IS VIDEO INVOLVING TWO THINGS. THE HETEROMER AND ALSO IN THE INTER-ACTION. WE LOOK FOR THIS BEHAVIOR IN DIFFERENT TISSUES, IN DIFFERENT MAMMALIAN CELL OR WITH THE RECEPTORS CONTAINING DIFFERENT RECEPTORS. WE ALWAYS FOUND THIS KIND OF INTER-ACTION BUT WE FIND INTER-ACTION WE FIND HETEROMER BECAUSE THE PROPERTY. HETEROMER. AMONG THOSE EXAMPLES WE HAVE STRIATAL AND HUMAN TRY A LITTLE TISSUE. THE DEMONSTRATION OF THE FUNCTIONAL SIGNIFICANCE. I TOLD YOU SEVERAL RESULTS THAT INDICATED THAT THESE TWO INTER-ACTIONS MIGHT COME FROM THE HETEROMERS. THESE EXPERIMENTS, WHAT I MEAN THAT EXPERIMENTS THAT DEMONSTRATE REALLY SUPPORT THAT THERE IS A FUNCTIONAL PROPERTY OF, IN THIS CASE, THE MSN THAT DEPENDS ON A2D 2 DIMERIZATION. THIS IS THE LABORATORY, WE SLICES OF MICE THAT EXPRESS GFP IN THE D2 RECEPTOR. THESE ARE DT MICE. WE CAN IDENTIFY THE D2 NEURON. WE APPLY AN MDA IN THIS PREPARATION THAT INDUCES VERY STRONG FIRING, COUNTERACTED BY THE D2 AGONIST. CGS WHICH IS A2 RECEPTOR AGONIST WHICH BY ITSELF WAS NOT DOING ANYTHING. COMPLETELY COUNTERACTED AFFECT OF THE D2 RECEPTOR AGONIST. LIKE D2 ANTAGONIST-LIKE BEHAVIOR. THIS IS DEMONSTRATING THE A DID-D 2 RECEPTOR. WHEN WE APPLY THIS PEPTIDE WHICH IS CORRESPONDS TO THE EPITOPE THAT IS PHOSPHORYLATED OF THE TAIL OF THE A2 RECEPTOR WE COUNTERACTED THE AFFECT OF CGS. INDICATING MOST PROBABLY THIS INTER-ACTION, RESPONSIBLE FOR THE FUNCTION. THE LAWS OF THE FUNCTION HERE INDICATES THAT THIS AFFECT IS RELATED TO A2-D 2 HETEROMERS TO MAKE A LONG STORY SHORT THERE ARE MANY EXPERIMENTS NOW THAT QUITE CONVINCINGLY SUGGEST AND WILL DEMONSTRATE THAT THE RECEPTOR RETRO MER IN THE INDIRECT IS RESPONSIBLE FOR MULTIPLE DEFICIT DEFECT FOR THE MODEL ACTIVATING, JUST TO -- I DON'T WANT TO GO IN DETAIL. ANOTHER AFFECT THAT WE SEE WITH MANY OTHERS MODELS. WHAT ABOUT THE SIGNIFICANCE OF THE RECEPTOR HETEROMER. WELL, WE FOUND IN THE LAB USING THE MODEL WE FOUND THAT FOR INSTANCE THAT THE A2A RECEPTOR ANTAGONIST SELECTIVELY D2 RECEPTOR AGONIST INDUCED -- BUT NOT ASK YET WHICH IS D1 SELECTED AGONIST THAT WAS DEMONSTRATING THAT IT CAN BE SELECTED IN THIS MODEL WHICH IS USED FOR PARKINSON'S DISEASE. THEN A GROUP THAT WORKS A LOT ON THIS ISSUE DEMONSTRATED THAT ALSO L-DOPA, WHICH CONVERTS TO DOPAMINE AGO TAKE VETS RECEPTORS VERY MUCH, CONTRA LATERAL IN -- WITH THE ADDITION OF ANTAGONISTS LOOK AT THE VERY STRONG POTENTIATION. ALL THESE PRE-CLINICAL DATA WE CAN USE A2 RECEPTOR AGONIST TO PROCEED TEN SHIITE THE AFFECTS OF L-DOPA TO INCREASE THERAPEUTIC INDEX. THERE HAVE BEEN SEVERAL THAT IS BASED ON THE REF STEP FOR HETEROMERS. I'LL JUMP TO ANOTHER HETEROMER WHICH IS ANOTHER A2A ALSO THE TRY @TIM. INVOLVED AS WELL THAT'S WHAT THE A1 RECEPTOR. NOW INSTEAD OF INHIBITING THE ABILITY OF DOPAMINE THE AGONIST TO BIND TO THE RECEPTOR NOW DOING THE SAME WITH THE STIMULATION OF -- DECREASES AFFINITY. SHOWING YOU JUST FEW EXPERIMENTS. THIS WOULD BE THE BINDING OF RP AN AGONIST TO FOR THE RECEPTOR. INCREASING CONCENTRATION ONLY WITH BUT NOT THE RECEPTORS. WHAT WAS NICE TO SEE THAT AGAIN THIS BY CHEMICAL FINGERPRINT CAN BE USED THAT THE SAME INTER-ACTION, SAME KIND OF BEHAVIOR COULD BE OBSERVED TO AIN'T CANADA THAT WE HAVE THIS A1 AND A2 RECEPTOR HETEROMER. BUT WHERE. THESE ARE RESULTS FROM THE GROUP, IMMUNOCYTOCHEMICAL STUDIES OF NERVE TERMINAL PREPARATIONS. THE A2A RECEPTOR YOU FIND IN GREEN. A1 IN BLUE. THEN MERGING OF THIS IMAGES GIVES WHERE YOU HAVE THE CO-LOCALIZATION. THIS INDICATED THAT AT LEAST A LITTLE BIT MORE THAN 50% OF TERMINALS THAT CONTAIN -- THAT MEANS TERMINALS HAVE BOTH A1 AND A2 RECEPTORS IN THE TRY @UM. AND EXPERIMENTS SHOWING IN THE SAME PREPARATION SHOWING THAT THE RECEPTORS ARE FROM COMPLEXES. TO MAKE LONG STORY SHORT GIVE YOU SOME OF THE RESULTS HERE FOR TO YOU SEE THE SIGNIFICANCE. I DON'T HAVE TIME TO GO IN DETAIL WITH OTHER EXPERIMENTS BUT WHAT KIND OF FUNCTIONAL SIGNIFICANCE DOES THIS HETEROMER HAVE. IT SEEMS THAT IT ACTS AS CONCENTRATION DEPENDENT SWITCH. LOW CONCENTRATIONS, THAT WOULD BE GLUTAMINE RELEASE AGAIN FROM THE SAME TYPE OF PREPARATIONS. NERVE TERMINAL PREPARATIONS THAT YOU STIMULATE WITH POTASSIUM. YOU INHIBIT THAT GLUTS MINI LEASE WITH LOW CONCENTRATIONS. THEN WHEN YOU INCREASE CONCENTRATION THERE'S SWITCH INSTEAD OF INHIBITING WHAT YOU GET IS RELEASE OF GLUTAMATE. IN THIS MECHANISM WITH THE INTER-ACTION -- BY MECHANISM IS THEN SINCE AS I SAID IT SEEMS THAT AROUND 50% OR A BIT MORE OF THE NERVE TERMINALS CONTAINING INTEROCEPTORS THAT WOULD HAVE DIFFERENTIAL EXPRESSION OF THIS HETEROMERS IN THE MSN. EXPERIMENTS AGAIN WITH D2 GFP MOUSE, THAT WAS IDENTIFY DOPAMINE D2 RECEPTOR CONTAINING THAT WOULD BE INCORRECT MSN AND RECEPTOR IS LOCATED IN THE D1, IN THE DIRECT MSN. RUN EXPERIMENTS AND IDENTIFIED DIRECT AND INDIRECT MSN. THAT WOULD BE IN THE LABORATORY. THIS EXPERIMENT SHOWED VERY CONVINCINGLY THAT ONLY IN THE DIRECT MSN WE COULD SEE MODULATION BY INTRODUCTION OF EITHER APPLICATION OF EITHER THE A2A AGONIST. OF THE POST SYNAPTIC CURRENTS. THAT WAS IN THE SENSE OF CGS INCREASING. AND SHARING THE ANTAGONIST DECREASING. THAT WAS ONLY IN THE DIRECT MSN NOT IN THE INDIRECT. ANALYSIS OF DIFFERENT PARAMETERS LIKE MEDIAN VARIANCE AND SO FORTH WOULD INDICATE THAT THESE WOULD BE MODULATION. THEN WE MADE ANALYSIS THAT RESULTS IN THE LAB THAT ALSO WOULD SUPPORT THIS SELECTIVE LOCALIZATION RECEPTORS IN THE GROUP OF TERMINALS. WE WOULD USE THESE EXPERIMENTS THAT WERE LABELING WITH AMINO -- FOR D1 RECEPTOR WHICH DIFFUSE STAINING. ALSO AT THE SAME TIME PARTICLE STAINING FOR THE RECEPTOR. WHAT YOU SEE HERE IS DIRECT MSN THAT EXPRESSES D1 BUT NOT A2 RECEPTORS. THIS WOULD BE DENDRITE THAT EXPRESSES A2 RECEPTORS, THE GOLD PARCEL BUT NO DIFFUSE THAT WOULD BE IMAGE OF DENDRITE OF MSN. A NICE PROFILE WHERE YOU CAN SEE NERVE TERMINALS THAT ARE MAKING IS IN NATURAL PARTICULAR CONTENT WITH SYMMETRIC SIGN UPS OF DENDRITIC SPINE. WE HAVE MANY MORE ELEMENTS LIKE THESE THAT WOULD BE IN INDIRECT MSN. THEN LOOK FOR IN VIVO MODEL TO STUDY THESE RECEPTORS. THEN WE IMPLANT IN THE STRAY YACHTS TARGETING AREA. WHAT WEIGH QUO SEE IS INCREASE IN GLUTAMATE RELEASE IN THE LATERAL TREE @UM, NOT IN THE MEDIAL TREE @UM, THAT WOULD NOT WORK. THE AFFECT OF THIS CORTICAL STIMULATION, GLUTAMATE RELEASE. WHAT WAS VERY INTERESTING ABOUT THIS EXPERIMENTS IS THAT IN ADDITION WHEN WE WERE PERFUSING WITH MS6 THE RECEPTOR ANTAGONISTS THOSE MOVEMENTS WHAT HAPPENS CORTICAL AREA WE CAN SEE MOVEMENTS OF THE -- IT DISAPPEARS. WE COULD ESTABLISH A MORE QUANTITY TATUM MODEL. WE'RE GOING TO DO HERE IS TO CORRELATE THE SIGNALS OF CORTICAL STIMULATION WITH THE ELECTROMYOGRAPHIC RECORDING FROM THE MASSES. HERE YOU WOULD SEE THE CORTICAL INPUT, THE RECORDING, EVERY TIME YOU SEE OPENING THE MOUTH. IS IT WORKING? THERE. YOU CAN SEE THIS IS INPUT, OUTPUT, THIS IS CORTICAL STIMULATION, THIS IS THE -- THEN STUDY CORRELATION BETWEEN THESE TWO SIGNALS AND VARIABLE THAT WE CALL POWER COLAKES COEFFICIENT. TURN OUT TO BE REALLY GOOD VIRAL KIND OF VERY STABLE FOR EACH ANIMAL EVEN BETWEEN ACROSS ANIMALS. THAT WOULD ALLOW US TO STUDY THE DIFFERENT COMPONENTS. WHAT WE FOUND, THESE ARE THE CONTROLS, FOUND THAT ANYTHING THAT WOULD DECREASE TRANSMISSION THROUGH THE PATHWAY LIKE A2 RECEPTOR ANTAGONIST. OR POST SYNAPTICALLY WITH THE D1 ANTAGONIST. VALUE FOR THE RECEPTOR. THEN WHAT WE DID STUDY EFFECT OF DIFFERENT TO RECEPTOR ANTAGONIST. IN THE MARKET THERE ARE MANY, WE CHOSE -- WE CHOSE SIX D2 RECEPTOR ANTAGONISTS AND COMPARED THE POTENT SEES TO -- ALSO COMPARE THAT TO POTENT SEES. WHAT WE FOUND IS THAT THERE WAS SOME COMPOUNDS LIKE -- THAT PRODUCES VERY NICE MOTOR ACTIVITY. DO NOT DECREASE PCC AND ALSO FOUND ANOTHER COMPONENT THAT WAS MORE INTERESTING FOR US THAT WAS SHARING 24416 THAT WAS VERY EFFECTIVE AT DECREASING WHILE WAS NOT PRODUCING MOTOR ACTIVITY. THAT WAS ALSO CHECKED WITH EXPERIMENTS THAT I TOLD YOU BEFORE. IMPLANTED ELECTRODES IN THE CORTEX. WE COULD SEE HOW THAT AFFECT WAS SIGNIFICANTLY COUNTERACTED BY SCH44216 ONE THAT WAS -- AND WAS NOT COUNTERACTED BY KW6002. THAT'S THE IDEA THAT I FORGOT TO SAY. THIS SYNAPTIC RECEPTOR. THEN IDEA WAS, WELL THEN WE SHOULD SHOW SOME TYPE OF CORRELATION WITH -- WE ASK OURSELVES IF DIFFERENCE IN POTENT SEES WOULD BE DUE TO DIFFERENCES IN -- THE RECEPTOR THAT IS PRESYNAPTIC WE HAVE VERY NICE CORRELATION WHEN WE STUDIES DISPLACEMENT OF THESE A2 RECEPTOR ANTAGONIST WITH 44246 IS PRESYNAPTIC A2 RESYSTEM FOR BIND TO -- FORMING HETEROMERS. WE WOULD COMPARE THE BINDING FROM CELLS ONLY CONTAINING A2A RECEPTORS AND OTHERS. WE FOUND THAT THIS WAS VERY POORLY TO THE CELL THAT WOULD BE EXPRESSED. IN COMPLETE AGREEMENT WITH THE DIFFERENTIAL POTENCY, OF THIS COMPOUND WOULD BE DUE TO DIFFERENTIAL ABILITY TO BIND TO -- DEPENDING ON THINK DIMERIZATION. CHEMICAL SIGNIFICANCE OF THAT WE'RE EXPLORING THAT. IN DRUG ADDICTION, THEY SHOW QUITE CONVINCINGLY THAT THE DIRECT PATHWAYS ARE IMPORTANT IN MODULATING STATEMENTS. WHAT WOULD BE -- IDEA THAT ANYTHING THAT WOULD DECREASE TRANSMISSION IN THE PATHWAY COULD BE OF THERAPEUTIC IMPORTANCE IN DUG ADDICTION. WE HAVE YEARS MODEL WHICH IS THE MONKEY MODEL IN THE THC. WHAT WE STUDY FIRST WAS THE ABILITY TO ANTAGONIST TO BLOCK, COUNTERACT ON ADMINISTRATION OF THC. THIS IS THE CONTROL THAT WOULD BE DOSE DEPENDENT INCREASE IN INJECTIONS PER SESSION. WHEN YOU PRODUCE DECREASE, SIGNIFICANT DECREASE IN ADMINISTRATION. WHAT HAPPENED WHEN WE INCREASED THE DOSE OF MSX WE SAW THE OP SIT AFFECT. A SHIFT TO THE LEFT. INDICATING WE THOUGHT THAT LOW CONCENTRATIONS, LOW DOSES OF MS6 WHAT IT'S DOING IS ACTING -- HIGHER DOSES IN A2D2 RECEPTOR THAT CAFFEINE POE TEN SHE YATES THE AFFECTS. SO WHAT WE'RE DOING IS EXPLORE THE AFFECT THAT HAVE SELECTIVE A2A ONE RECEPTOR ANTIS. SO FAR DIFFERENT DOSES OF -- ONLY THING THEY'RE DOING -- FEIGNLY MY LAST RECEPTOR HETEROMER. I INTRODUCE YOU FAMILY OF DOPAMINE RECEPTORS THAT YOU PROBABLY KNOW THAT THEY ARE DIVIDED IN D1 AND D2 RECEPTORS. D1 RECEPTOR -- WHAT IS KNOWN IS THAT MANY STUDIES SHOWING HETEROMERIZATION BETWEEN MANY TYPES OF RECEPTORS. ALSO SAME GROUP, WE HAVE SOME RESULTS ON -- ALSO BETWEEN D2 AND D3 THERE WERE NO STUDIES INCLUDED IN THE RECEPTOR WE THOUGHT MAYBE WOULD BE INTERESTING BECAUSE IT HAS VERY -- FUNCTIONS HAVE BEEN SHOWN VERY MUCH. IT BOUND WITH A LOT OF AFFINITY, CLOSE. ALSO LIKE FEW YEARS LATER WAS FOUND THAT THE D4 DOPAMINE D4 RECEPTOR GENE CONTAINS LARGE NUMBER. MOST INTERESTING ARE THE ONES THAT ARE LOCATED IN X AND 3 WHICH GOES FOR 13 OR D4 RECEPTOR THIS CONSISTS OF NUMBER OF REPEATS. THAT WE HAVE REPEATED THE GOAL FROM TWO TO 11 IN HUMANS. THESE ARE FREQUENCY, GLOBAL FREQUENCIES, MOST COMMON IS D44 WITH 65%, 65% OF GLOBAL FREQUENCY. THAT'S ABOUT 10%. NOT REALLY STUDIES SHOWING ANY FUNCTIONAL DIFFERENCES BETWEEN THESE DIFFERENT VARIANCE OF D4 RECEPTORS. WHAT WAS IMPORTANT IS THAT ABOUT VARIANCE AT LEAST IT'S QUITE CLEAR, EPIDEMIOLOGICAL STUDIES HIGH PREVALENCE. WE STARTED BY TESTING DIFFERENT CELLS AGAIN, POSSIBILITY OF HETEROMERIZATION BETWEEN D44 AND D47 THE THREE MAIN VARIANCE IN D2 RECEPTORS, AND D1. WE COULD FIND, THAT WAS REALLY UNEXPECTED WE COULD FIND BETWEEN THEM THERE WAS POSSIBILITY THAT WOULD MAKE SENSE. THE D47 WAS NOT DIMERIZING. ALSO NEITHER DID ANY OF THE VARIANCE WERE HETEROMERIZATION. THERE WAS VARIABLE DEPENDENT OF D4. WHAT WE ADDRESS WAS ACTIVATION. THIS IS PHOSPHORYLATION, WHAT WE FOUND THAT D4 RECEPTOR AGONIST THOSE DEPEND ON PRODUCE ACTIVATION AND PHOSPHORYLATION. INTERESTING FINDING WAS THAT THIS IS -- THE D2 RECEPTORS WHICH IS ONE THAT IS NOT -- THE CONTROL, IT WOULD BE SHIFTED TO THE LEFT WITH THE ADDITION OF THE D2. THERE WOULD BE D2-D4 RECEPTOR. OR D2 AND D44. BUT IT WOULD NOT HAPPEN WITH THAT WOULD BE TRANSECTED. THIS LAW OF FUNCTION IS ASSOCIATED WITH LOSS OF -- SUPPORTED STRONGLY THAT THIS D2-D4 INTER-ACTION IS BY CHEMICAL PROPERTY OF THE RECEPTOR HETEROMER. THEN WE WANTED TO SEE DEMONSTRATE HETEROMERS IN THE BRAIN. AND IN THE LAB THEY CREATED A KNOCKING MOUSE, VERY INTERESTING, THAT INSTEAD OF HAVING THE D4 MOUSE WE -- BUT ALSO D47 HUMAN WOULD NOT -- THERE WERE CONTROLS WE NEEDED TO REACH A CONCLUSION I'LL TELL YOU LATER. SAME KIND OF CORRELATION THAT DID 2S MOUSE WOULD LOSE ALSO THAT BIOCHEMICAL PROPERTY. WE DON'T HAVE HIT REAR SAKES YET. WITH THE D4 WOULD SHOW THAT KIND OF INTER-ACTION I WAS TELLING YOU BEFORE. VERY NICELY NEITHER THE D4 RECEPTOR AGONIST, OR D2 RECEPTOR WERE NOT SELECTED. THERE IS TREND BUT NO SIGNIFICANT ACTIVATION. VERY IMPORTANT TO SEE THAT THE CURRENT ACTIVATION OF D2 AND D4 RECEPTORS PRODUCE A VERY STRONG DOSE DEPENDENT INCREASE MECHANISM. THIS COULD BE ALREADY DEMONSTRATED -- DEMONSTRATION OF THE HETEROMERS. ANY DOUBT IN THE D47 MOUSE THERE WAS NO INTER-ACTION WHATSOEVER. WHAT THE FUNCTION OF SIGNIFICANCE LOCATED D4 RECEPTOR HETEROMERS, D4 RECEPTORS IN THE TRY @UM, WE SHOW THE AFFECTS THERE, D4 RECEPTORS THEY WERE PREVIOUSLY FOUND TO BE LOCATED IN TERMINALS OF CORTICAL NEURONS. THIS COMES WHERE STUDY BINDING OF D1 RECEPTORS AND AFTER ABLATION OF THE FRONTAL IT WOULD BE SIGNIFICANT DECREASE OF D4 RECEPTOR BINDING. WHAT THE FUNCTION, THERE WAS NOTHING KNOWN ABOUT THE FUNCTION OF D4 RECEPTORS. THIS IS DONE IN THE LABORATORY WHAT STUDY IS AFFECT OF POTASSIUM, GLUTAMINE POTASSIUM AND AFFECT OF D4 RECEPTOR AGONIST THAT SHOWED VERY SINCINGLY DECREASE GLUTS MEN. THAT WAS COMPLETELY COUNTERACTED TO THE ADMINISTRATION OF D4 SELECTIVE ANTAGONIST RESULT CELLS. I'M NOT SHOWING RESULTS BUT THAT WAS SELECT FOR GLUTAMATE NEITHER DOPAMINE NOR GAB BEEN WAS MODULATED BY THE RECEPTORS. A SELECTIVE MODULATION. THEN WHAT WE FOUND -- WHAT WE FOUND IN HIS LAB FOUND IS THAT THE RESULTS OF D2 RECEPTORS, FINDING OTHER EXPERIMENTS AND OTHER STUDIES CAN MODULATE. SO WHAT I'M SHOWING NOW HERE IS DOPAMINE RELEASE, INCREASING CONCENTRATIONS OF DOPAMINE -- DOPAMINE RELEASE OF GLUTAMINE. THIS IS THE RESPONSE CURVE AND THAT COUNTERACTED PARTIALLY BY DOPAMINE TO D2 RECEPTOR ANTAGONIST A LITTLE BIT MORE BUT D4 ANTAGONIST, THAT'S WHAT I WANT TO YOU SEE COMPLETELY BY CO-ADMINISTRATION OF D4 AND D2 RECEPTOR ANTAGONIST. INDICATING BOTH ARE INVOLVED. WHAT WAS EVEN MORE IMPORTANT WAS THAT WE COULD SHOW THAT THOSE RESPONSE -- THIS IS D4 RECEPTOR ON INHIBITION. IS SHIFTED TO THE LEFT BY CO ADMINISTRATION OF D2 RECEPTOR AGONIST. ALMOST MY LAST SLIDE. THESE RESULTS START TO MAKE UNDERSTAND PROBABLY A LITTLE BIT MORE OF WHAT HAPPENS IN THE -- OR AT LEAST WHAT MECHANISM BEHIND THERAPEUTIC EFFECTS OF STIMULANTS. CAN ALL UNDERSTAND WHY MORE RESISTANCE TO THE TREATMENT IN PATIENTS THAT HAVE THE VARIANT. THAT WILL BE THE CONCLUSIONS SLIDE BUT I WANT TO CHALLENGE THE AUDIENCE, I WOULD LIKE TO FINISH NOT WITH A CONCLUSION BUT WITH A NEW TITLE. I WANT TO CALL IT RESTEP FOR METRO MERS AN OPENING TO A BETTER UNDERSTANDING OF RECEPTOR PHYSIOLOGY AND PHARMACOLOGY I WOULD LIKE TO FINISH WITH THESE THREE STATEMENTS. BOTH FROM PHYSIOLOGICAL AND PHARMACOLOGICAL POINT OF VIEW. SHOULD NOT BE CONSIDERED ANY MORE -- BOTH FROM A SYS LOGICAL AND PHARMACOLOGICAL POINT OF VIEW, RECEPTORS SHOULD NOT BE CONSIDERED ANY MORE AS SINGLE FUNCTIONAL UNITS. TWO, RECEPTOR HETEROMERS UNCOVER A PREVIOUSLY UNFORESEEN VAST NUMBER OF NEW POSSIBLE SUBPOPULATIONS OF GPCR SUBTYPES, WITH SPECIFIC NEURONAL LOCALIZATIONS AND FUNCTIONS. THEIR LIGAND SELECTIVITY IMPLIES THAT RECEPTOR HETEROMERS CONSTITUTE POTENTIAL NEW TARGETS FOR DRUG DEVELOPMENT. THEN I HAVE IMPORTANT COLLABORATORS AND INITIAL THAT I TRIED TO CITE EVERY TIME THAT I WAS SHOWING EXPERIMENTS. THANK YOU VERY MUCH. [APPLAUSE] >> I HAVE A QUESTION ABOUT YOUR LAST -- [ NOT AUDIBLE ] >> THE GLUTAMINE RECEPTOR. >> THEY RANK ORDERED RELATIVE BINDING OF THE -- THEY SHOWED RATIO OF THE -- IS IT SIMILAR. [ NOT AUDIBLE ] >> CAN YOU REPEAT THAT? THIS IS THE BROADCAST. >> I KNOW, THAT'S A LONG QUESTION. WHAT QUESTION WOULD SHE LIKE. >> IT'S BASICALLY THE ANSWER -- THE QUESTION IS, CITING ANOTHER WORK OF WORK RECENTLY PUBLISHED, THEY WERE SHOWING DIFFERENTIAL AFFINITY FOR THE -- TWO DIFFERENT ANTI-PSYCHOTICS THEY FOUND KIND OF A NICE CORRELATION DEPENDING ON WHICH ONE WOULD BE CLINICALLY MORE ATYPICAL, IF I REMEMBER CORRECTLY. IN THAT CASE WHAT HAPPENS -- REALIZE THAT EVERY SINGLE HETEROMER IS A NEW STORY. BUT IN GENERAL IT DEPENDS ON WHAT WE'RE TALKING ABOUT. IN MY CASE WE'RE FOCUSING IN THE AFFINITY OF ONE OF THE HETEROMERS AND THAT'S WHAT I BELIEVE A LOT IN TO TARGETING BECAUSE THAT'S WHAT I WANT TO -- AS MUCH AS POSSIBLE. IN THIS CASE IT WAS JUST THE A2A RECEPTOR COULD HAVE GONE TO THE D2 RECEPTOR AS WELL. WE DID NOT STUDY DIFFERENCES IN THE BINDING OF THE D2 RECEPTOR LIGANDS. WHAT YOU'RE SAYING THE SAME LIGAND, IF I UNDERSTAND CORRECTLY WAS BINDING TO THE TWO -- THAT'S ANOTHER EXAMPLE. BUT IN MY CASE I WAS JUST FOCUSING ON ONE OF THE ELEMENTS. FOR ME THE SIMPLE WAY -- SIMPLEST WAY TO GO JUST FOCUS ON ONE AND SEE IF WE CAN TARGET IT BETTER WITH OTHER LIGAND. BUT THEY ARE DIFFERENT APPROACHES. ALSO I'M NOT SO SURE -- THAT WAS BINDING. MOSTLY NOT LOOKING AT DIFFERENT AFFINITIES ON BINDING AS FAR AS I REMEMBER. THERE WERE MOSTLY LOOKING AT THE SIGNALLING. HOW THE FINAL RESULT OF THE AFFECT, THERE WAS THE EFFICACY IN TERMS OF SIGNALLING. WE ARE FOCUSING MORE IN BINDING. YOU CAN GO TO THE END, I WAS SHOWING THIS DIFFERENT BIOCHEMICAL PROPERTIES OF HETEROMERS ALL THESE THINGS CAN HAPPEN, CHANGES IN LIGAND SELECTIVITY, CHANGES IN SIGNALLING ANYTHING CAN HAPPEN. BUT I THINK THAT WHAT -- SEE IF I CAN HELP WITH THAT. WHAT I WANT FIRST TO FIND RECEPTOR HETEROMER THAT CAN BE A TARGET THEN STUDY ALL POSSIBILITIES, ALL THE BIOCHEMICAL PROPERTIES THEN FIND WAY TO DISSECT COMPARED TO RECEPTORS FROM SOMEWHERE ELSE. >> I WAS WONDERING JUST WHAT YOUR THINKING IN WHEN ME OF HETERODIE MANNED WHERE THE ENDOGENOUS LIGAND, LIKE DOPAMINE, HETERODIMERS IF THE IDEA THAT YOU THINK THAT RECEPTORS ARE PREFORMED AS HETERODIMERS OR IS THE IDEA THAT WHEN THEY BIND ONE OF THE HIGHER AFFINITY HAVE POSSIBILITY OF FORMING DIMER AFTER THAT, IF I COULD FOLLOW AS SECOND PART THAT HAVE QUESTION IS, VERY OFTEN THESE RECEPTORS ARE SEEN TO BE INTRACELLULAR, DO YOU HAVE ANY EVIDENCE THAT ONE MAY HELP THE OTHER ONE. >> AGAIN, DIFFERENT RECEPTOR METRO MERS, DIFFERENT FUNCTION, DIFFERENT BEHAVIORS, IT SEEMS THAT FOR MOST OF THE EXAMPLES THAT WE KNOW IN THE FIELD THAT -- THEY GO ALREADY PREFORMED TO THE MEMBRANE, TO THE PLASMA MEMBRANE. IN THAT RESPECT -- THERE ARE EXAMPLES, QUITE GOOD EXPERIMENTS SHOW SOME HETEROMERS OR LIGAND PROMOTES -- THAT'S STILL POSSIBLE. AGAIN WHAT THE RECEPTOR YOU'RE INTERESTED IN LET'S STUDY THEM. BUT IN GENERAL THEY WILL BE PREFORMED. >> WE'RE VERY INTERESTED IN MULTI-VEIL ANT LIGANDS. CAN YOU DESCRIBE ANY EXPERIMENTS EITHER WITH THE DOPAMINE SYSTEM OR ANY OTHER HETERO-RECEPTORS WHICH YOU GET A SELECTIVITY OR SELECTIVE ENHANCEMENTS IN A PARTICULAR HETEROMERIC PAIR. WITH MULTI-VEIL ANT. >> ONE PART OF THAT COMPOUND WOULD BE AN AGONIST FOR THE D2 AND OTHER PART WOULD BE ANTAGONIST FOR AT A 2A. THAT IS ONE WAY TO FIND COMPOUNDS. PROBLEM IT HAS NOT TO MY UNDERSTANDING SHOWN THAT YOU ARE REALLY BINDING TO THE TWO POET MERS AT THE SAME TIME. THEY ARE SUPPOSED TO BE TRANSIENT ENTITIES FOR WHAT WE START TO UNDERSTAND NOW. I THINK, I HAVE HEARD CRITIQUE THAT THE PHARMACOKINETICLY CAN BE BAD. IT WOULD BE DIFFICULT TO ADMINISTER. YOU PROBABLY KNOW BETTER THAN ME ABOUT THAT. IN PRINCIPLE THEY'RE GOOD TOOLS I WOULD SAY. BUT I'M NOT SO SURE THEY COULD BE GOOD THERAPEUTIC TOOLS. >> I HAVE TWO QUESTIONS. FIRST I WONDER IF ANY OF THESE HAVE BEEN GLIAL CELLS, ASTRO CITES IN PARTICULAR. ANY BETWEEN A CLASSICAL NEUROTRANSMITTER AND A PEPTIDE TRANSMITTER. >> RECEPTOR WITH THE CLASSICAL. >> WHERE YOU HAVE HETEROMER OF DOPAMINE AND ONE OF THE OPIATE RECEPTORS. >> THERE ARE. >> DOPAMINE -- >> THERE ARE. WE ARE WORKING NOW, WE DESCRIBE, IN THE HIPPOCAMPUS WE'RE LOOKING FOR THAT NOW. EVEN FARTHER, WE CAN FIND HETEROMERS THAT INCLUDE GPCRs AND LIGAND CHANNEL, IS THAT IS DESCRIBED, FOR INSTANCE, D1 -- ONE OF THE FIRST, BY THE WAY, IN "NATURE AND SCIENCE" PAPERS, D1 WITH -- AND R1 THEN D5 WITH THE GABA A. IT'S NEVER ENDING. ALL THE POSSIBILITIES ARE THERE. >> AND ARE THEY IN GLIAL CELLS? >> THE GLIAL CELLS. I'M SURE THEY ARE. WHAT I TEND TO BELIEVE IS THAT RECEPTORS THAT CAN BE FORMING HETEROMERS IN RESPECTIVE CELLS ARE CO-LOCALIZED FOR GOING TO FORM IN GENERAL. IN THE ASTRO SITES HAVING DESCRIBED, WE DESCRIBE THE RECEPTOR HETEROMER THAT COULD BE ONE. MANY OTHERS. >> OKAY. THANK YOU VERY MUCH FOR STARTING OFF OUR SERIES. [APPLAUSE]