SO IT'S MY GREAT PLEASURE TO INTRODUCE TODAY'S SEMINAR SPEAKERS, MING GOU FROM UCLA. SO I JUST GOT A NOTE THIS MORNING THAT MING AND I GRADUATED FROM MEDICAL SCHOOL--NOT IN SAME SCHOOL BUT NEXT TO EACH OTHER, IT'S ONLY A FEW MILES AWAY IN SHANGHAI BACK TO THE 1980S AND MIDDLE 1980S. SO MING GOU RECEIVED HIS M. D. DEGREE IN SHANGHAI CHINA AND THEN SHE CAME TO U.S. AND PURSUED Ph.D. DEGREES AT UCSF. AND SEE DID IT AT THE EXTREMELY VERY SUCCESSFUL [INDISCERNIBLE] WORK AND I JUST LISTED THE 1--MENTION 1 OF THE STUDIES AND SHE CONTRIBUTES AND IDENTIFY A NOVEL AND SIGNALING, NOTCHED SIGNALING PATHWAYS WHICH WAS CRITICAL FOR REGULATING THE CELL DIVISION, WITH STEM CELL DEVELOPMENT. AND AFTER Ph.D. RESEARCH AND THEN SHE DECIDED TO RETURN TO THE CLINICAL [INDISCERNIBLE] BY FINISHING THE NEUROLOGY RESIDENCE TRAINING IN SURVEYS SON AND FOLLOWED BY FELLOWSHIP TRAINING IN DEGENERATIVE DISEASES IN THE UCLA AND BEFORE SHE BECAME AN ASSISTANT PROFESSOR AT THE DEPARTMENT OF THAT YOU'ROLOGY. SO--NEUROLOGY. SO ABOUT 3 OR 4 YEARS AGO, MING WAS PROMOTED TO TENURE. AND DURING HER CAREER IN? MING RESILIENCE CEIVED A NUMBER OF SCIENTIFIC---RECEIVED A NUMBER OF AWARDS WHICH WAS FROM THE NEUROSCIENCE AWARD, AND NEUROLOGICAL SOCIETY AWARD. AND SHE IS THE ELECT MEMBER AS AMERICAN NEUROLOGICAL ASSOCIATION. SO MING ALSO ACTIVELY SERVED TO THE NEUROSCIENCE COMMUNITIES AND SO, I JUST LIST A FEW, AND SHE WAS A REGULAR MEMBER [INDISCERNIBLE] FOR SEVERAL YEARS AND SHE RECENTLY, WITH THE BSC AND BOARD OF SCIENTIFIC COUNSELOR IN OUR INSTITUTE IN IDEAS AND SHE ALSO SERVED AS THE ASSOCIATE DIRECTOR FOR TRAINING FOR MEDICAL TRAINING BETWEEN UCLA AND CAL TECH IN THE PROGRAM. SO MING'S LAB IN THE PAST DECADE MADE A NUMBER OF IMPORTANT CONTRIBUTIONS TO OUR UNDERSTANDING FOR THE 2 MOST COMMON NEURODEGENERATIVE DISEASES, INCLUDING ALZHEIMER'S DISEASE AND PARKINSON'S DISEASE, I'M SURE MING WILL TALK TODAY ABOUT THESE STORIES. AND MING, BASICALLY USING DROSOPHILA AS A MODEL TO PROVIDE EVIDENCE TO SHOW THE UNDER SCORE AND IMPORTANCE THAT THE MITOCHONDRIAL DYSFUNCTION AS A CENTRAL MECHANISM FOR THE PARKINSON'S DISEASE AND PATHOGENESIS. THEIR STUDIES OPEN THE DOOR FOR THE STUDY OF THE INTERNATIONAL CLASSIFICATION TERNAL SIGNALING AND THE [INDISCERNIBLE] SIGNALING AND MITOCHONDRIAL FUNCTIONS IN THE DISEASE MECHANISM. SO PLEASE JOIN ME TO WELCOME MING. [ APPLAUSE ] >> THANK YOU SO MUCH FOR YOUR GENEROUS INTRODUCTION, I JUST WANT TO SAY THAT NIH IS 1 OF MY FLAIFORT PLACE TO GO ON AND THERE ARE THESE THINGS GOING ON AND SO NOT ONLY COMING HERE FOR SERVICE BUT FOR SCIENCE. I HAVE SEVERAL WONDERFUL COLLEAGUES HERE, ALWAYS FEEL VERY SIMULATED AND EXCITED WHEN I COME TO THIS WONDERFUL PLACE. SOILY MENTIONED I'M A NEUROLOGIST, SEEING PATIENTS WITH NEUROLOGICAL DISEASES AND ALSO RUN A RESEARCH LAB STUDYING MOLECULAR MECHANISMS OF THE 2 MOST COMM3Eb/9 NEURODEGENERATIVE DISEASES AND I TODAY I WILL TELL YOU THE STORY RELATED TO PARKINSON'S DISEASE. >> COULD THE LIGHTS BE DIMMED A LITTLE. SO THESE ALL HAVE THINGS IN COMMON AND AFFECTING 5% OF PEOPLE OVER THE AGE OF 80, SO WHAT ARE THE SYMPTOMS OF OF DISEASE, WHAT I USUALLY WHEN I GO TO VISIT PLACES, PEOPLE ALWAYS WANT THE TO TAKE A LOOK AT IT, THIS SUBJECT WORKING OUT SO MAYBE SHY JUST PLAY THESE 2 PATIENTS SIDE BY SIDE, SO WHAT WE DO IS ASK THEM TO TAP THE FINGERS AS FAST AS YOU CAN AND THEN YOU SEE IF YOU EVER HAVE A DEFICIT AND THEN LOOKING AT FACIAL EXPRESSION, WE SAY POSITIVE FACIAL EXPRESSION, LOOK AT FACIAL EXPRESSIONS OR MASKED PHASING, AGAIN, SLOWNESS IN MOVEMENT, BECAUSE YOU'RE SUPPOSED TO DO IT REALLY FAST AND THEN ON THE RIGHT HAND SIDE WE ASK PATIENT TO WRITE HIS NAME AND THEN DRAW A SINGLE LINE IN BETWEEN THESE SPIRAL LINES ANDEN IN THE LEFT-HAND SIDE NOTICE THERE IS RESTING TREMOR RELATED TO HIS HAND AND DIFFICULTIES WITH GAIT WHICH IS WALKING AND I WANT TO TELL YOU IS ALSO SLOWNESS OF MOVEMENT, RESTING TREMOR, MUSCLE STIFFNESS, ALL THESE SYMPTOMS TOGETHER IN A CLINICAL SENSE, WE CALL THEM MOTOR SYMPTOMS AS THEY RELATE TO MOTOR. LET'S SEE HOW THIS GENTLEMAN DID, NOT VERY GOOD, VERY, YOU KNOW--NOT STRAIGHT LINE. AGAIN WE ASK THE PATIENT TO PERFORM A TASK THAT IS TO POOR WATER FROM 1 GLASS INTO ANOTHER. TREMENDOUS DIFFICULTY. AS YOU CAN SEE THIS WOULD FOR SURE COMPROMISE HIS ACTIVITIES OF DAILY LIVING SUCH AS DRINKING OR EATING AND I WANT TO DRAW YOUR ATTENTION THAT THIS IS NOT THE END STAGE OF PARKINSON'S DISEASE, THIS IS ONLY THE MIDSTAGE IN THE END, IT'S A DEBILITATING PROGRESSIVE DISEASE IN THE END, THEY ALL ENDED UP BEING BEDRIDDEN. WHEELCHAIR BOUND. SO THE PATHOLOGY OF PARKINSON'S DISEASE IS ROUGHLY KNOWN. FOR LONGEST TIME, PEOPLE KNOW THAT THERE ARE THESE NEURONS LOCATED IN MIDBRAIN WHICH IS PART OF THE BRAIN STEM, CALLED DOPAMINERGIC NEURONS AND SUBSTANTIAL NIGRA, SO IF YOU HAVE YOUR HEAD AND IF YOU WERE TO CUT THIS AND FLIP IT, THIS AREA IS A SUPERIMPOSED BRAIN STEM AREA IN THIS MRI REGION AND HERE IS JUST THE FRONT THIS, IS YOUR EYE BALLS AND THIS IS YOUR NOSE AND THIS IS THE BACK, SO THE PIGMENTED NEURONS DIMINISH AS YOU GENERATE, AS YOU DEVELOP PARKINSON'S DISEASE. SO THEN, ISN'T THAT EASY, ANOTHER TREATMENT IS JUST PARKINS--DEPRIVATIONA MIN REPLACEMENT THIS, IS INDEED WHAT HAS BEEN EXCUSED IN CLINICAL WORLD. HOWEVER THERE ARE SEVERAL PROBLEMS, 1 THING IS THESE PATIENTS, ONCE YOU SEE A PATIENT, THEN YOU WOULD REALIZE IT. PATIENTS ALSO DEVELOP A VARIETY OF NONMOTOR SYMPTOMS INCLUDING PSYCHIATRIC SYMPTOMS SUCH AS DEPRESSION, COMPULSIVE BEHAVIORS AND ANXIOUS AND THEN OTHER SYMPTOMS. SOME OF THEM EVEN OUTSIDE OF THE NERVOUS SYSTEM, SO 30% OF PEOPLE AT LEAST DEVELOP DEMENTORSHIP SKILLIAL. SO WHAT DOES THAT TELL US? IT'S NOT THE ONLY STORY BECAUSE ONLY SOME OF THE MOTOR SYMPTOMS CAN BE SUCCESSFULLY TREATED BY DOPAMINE ONLY AT THE EARLY STAGE, FOR INSTANCE, GAIT AND STABILITY IS NOT GOOD FOR TREATMENT. SO THERE ARE OTHER SYMPTOMS MORE DEBILITATING AND WITHOUT GOING DETAILS THIS IS SUPPORTED BOY THE PATHOLOGICAL FINDS AND THEN A BARAK GROUP LOOKED AT THE PATHOLOGY OF THE BRAG AND FOUND OUT THAT THAT THIS START INDEED THE LOW BRAIN STEM, BY THE TIME IT REACHES THE SUBITANTIA NIGRA, IT'S REACHED THIS STAGE, SO THIS IS BEYOND DOPAMINERGIC NEURONS AND WE NEED TO DEVELOP A UNDERSTANDING OF THE MECHANISM TO DEVELOP NEW TREATMENT STRATEGY. FORTUNATELY, FOR THE PAST 15 YEARS, A SERIES OF GENE VS BEEN CLONED. I MEAN THERE ARE PLUS, AND MINUSES, HERE IS CERTAIN LISTS. THIS IS WHAT ALMOST EVERYBODY AGREED, SO, THESE ARE THE 2 NEW GENES. SO TOGETHER, THEY AFFECT ROUGHLY ABOUT 10% OF THE CASE. IT'S NOT A SMALL NUMBER AND TODAY I'M GOING TO TELL YOU, SOME OF THE STORIES REGUARDED TO PINK 1 AND PARKINS, AND LRRK2, USING DROSOPHILA AS A MODEL. SO, WHY STUDY THE CASE AND THIS CLINICAL CASES, THESE ARE SIMILAR TO A SPORADIC CASE, MAYBE HETEROGENEITY BUT BY FINDING THE FAMILIAL GENE MECHANISM, IT MAY BE SIMILAR. SO WE STARTED OUT THIS PROCESS BY LOOKING AT A PROTEIN CALLED PINK 1. P10 INDUCED KINASE, AND I WANT TO ACKNOWLEDGE THERE'S A LOT OF PROGRESS MADE BY MARK, RICHARD AND AMONG OTHER COLLEAGUES, MANY, SO ANYWAYS, WHAT IT IS IS THIS PROTEIN, WHEN IT JUST CLONED, I GOT EXCITED BECAUSE THIS WAS THE PROTEIN, IT'S VERY UNIQUE, HAS A MITOCHONDRIA TARGETING SEQUENCE AND HAS A SERIES OF KINASE, SO THESE 2 STRUCK ME AS BEING REALLY IMPORTANT. WORN IS THE MIGHT O KONDCHONDRIA TARGETING SEQUENCE SO WE KNOW IN THE PAST, THERE ARE IS STUDIES LINKING MITOCHONDRIA DISTONKS TO THE DISEASE, YET THOSE ARE MORE TOXIN MODELS, NOT REALLY DEGENERATION MODELS, BUT NONETHELESS IT'S JUST INTERESTING, AMONG ALL THESE LISTS OF GENES, THIS IS THE OLDSMOBILE 1 THAT HAS THE TARGETING SEQUENCE. AND I WAS SURPRISED TO SEE THIS, SO THIS IS JUST VERY INTRIGUING. TO START THIS PROCESS WE GENERATE A DROSOPHILA DELETION, MULTIPLE DELETIONS, THIS 1 IS NOT PUBLISHED BUT IT'S REMOVED ALL THE CODING REGIONS, NONE OF THE SURROUNDING GENES AND WE'RE COMPULSIVE, WE ALSO MADE A PINK 1 RESCUE GONE--GEAN ORDER OF MICRONSIC RUSE CUED FROM THIS GENOMIC RESCUE TRANSGENE. WHAT IS THE PHENOTYPE? IT'S THESE NONMUTANT FLIES THAT ARE VIABLE BUT THEY HAVE CERTAIN DEFECTS. SO INCLUDING DOPAMINEERGIN NEURONAL DEFECT ANDvXC STRIKINGLY, THEY HAVE MUSCLE DEFECT AND MALE STERILITY DEFECT AND THEY HAVE STRESS SENSITIVITY AND SHORTEN LIFE SPAN AND ALL OF OF WHICH CAN BE ATTRIBUTED TO MITOCHONDRIA DYSFUNCTION AS I'LL TELL YOU LATER. SO, FOR THE TIME SAKE WE'RE GOING TO FOCUS ON TODAY FOR MUSCLE AND THE GERM LINE BECAUSE THESE 2 TISSUES RELY HEAVILYOT MITOCHONDRIA TO MEET THEIR ENERGY DEMAND. SO THIS IS WHAT WE SEE. SO HERE'S WILD-TYPE. THIS IS PINK 1 MUSEUMITANT. LIGHT MICROSCOPIC VIEW THIS, IS THE ELECTROMICROSCOPY VIEW. SO IN WILD-TYPE WHAT YOU SEE IS A MUSCLE SECTIONS LOOKING AT MUSCLE MATURITY AND THEY'RE HIGHLY ORGANIZE WIDE REGULAR BENDING WHERE THE PINK 1 MUTANT USES THESE IN DEGENERATION. AND THEN UNDER THE ELECTRON MIRROR MICROSCOPY, SO HERE IS A BLOW UP VERSION, THIS PARTICULAR 1, ACTUALLY A RESCUE, PEOPLE WOULD RESCUE SO WE SEE IN THIS KIND, YOU HAVE THIS DENSELY PACKED MUSEUMITANT ARE LARGELY BROKEN, AND SOME OF THEM LOOKS HORRIBLE THIS, IS A 14 DAY OLD--SO IT'S NOT--IT'S NOT SUBTLE TO SAY THERE IS A PHENOTYPE. SO WE ALSO STAIN THIS. THE THERE ARE INDICATIVE THESE MUSCLES ARE UNDERGOING CELL DEATH. SO WHAT WE ALSO DID WAS MADE A FLY ANALOGOUS TO PINK 1 MUTANT, SHOW IS MINIPATIENT FLIES SO TO SPEAK. THEY SHOW SIMILAR TYPES TO PINK 1 NULL, THIS HAD IS SIMILAR TO THE MEDIATED PD IS A LOSS OF FUNCTION DISORDER. SO 1 EXCITING THING IS THAT WE FOUND THAT THEY ACTUALLY SHOW A SIMILAR PHENOTYPE TO ANOTHER MUTANT, CALLED PARKINS THAT WAS PREVIOUSLY STUDIED BY OTHER FLY LABS AND THEN THIS PROVIDES US AN OPPORTUNITY FOR US TO ADDRESS THIS LARGE QUESTION, FOR THE FAMILIESIAL PARKINSON'S DISEASE, DO THEY ALL IMFOANCH A MULTIPLE PATHWAY THIS, IS JUST 1 PIECE OF IT, SO IN DROSOPHILA WE'RE ABLE ON DO A FAIRLY RIGOROUS STUDY WHICH I'M GOING TO GIVE YOU A FLAVOR OF, SO DO THESE FUNCTIONS IN THE TOGETHER IN THE SAME PATHWAY. WELL WE INITIALLY TAKE IN A PINK 1 MUTANT BACKGROUND AND OVEREXPRESSED PARKIN IN THE BACKGROUND LOOKING AT THE MUSCLE, SO HERE'S THIS, YOU SEE, YOU SEE BROKEN, THIS PHENOTYPE IS A BIT WEAKER, BECAUSE THIS 1 IS 2 DAY OLD, PREVIOUS 1 WAS 14 DAY OLD AND WHEN YOU SEE, THIS BACKGROUND, THERE'S 4 MITOCHONDRIA HERE AND 3 OF THEM ARE RESCUED AND YOU NEVER SEE THIS PHENOTYPE IN PINK 1 MUTANT. SUGGEST THANKSGIVING THIS PHENOTYPE CAN BE RESCUED WHICH IS VERY STRIKING. SO THERE ARE ACTUALLY 2 WAYS I WANT TO JUST DRAW THIS ATTENTION BECAUSE PEOPLE USUALLY, IN THE LITERATURE CAN MISINTERPRET CERTAIN FACTS. SO FOR JUST OVER EXPRESSION RESCUE, THEY'RE ARE 2 WAYS OF INTERPRET THANKSGIVING DATA, 1 IS A LINEAR PATHWAY MODEL, THAT IS ACTIVATED PARKIN AND MIGHT MITOCHONDRIA AND ACTUALLY, OF COURSE, IN THE PINK 1 MUTANT IF YOU SUPPLY PARKING YOU HAVE A RESCUE. HOWEVER, THIS--HOUR DATA IS COMPATIBLE WITH A PARALLEL MODEL. WHAT DOES THAT MEAN? THE PARKIN ACTS IN PARALLEL WORKS ON ACT ON THE COMMON TARGET IN THE PINK 1 MUTANT IF YOU'RE OVERDRIVE THANKSGIVING BRANCH, IN YOU CAN,A CHIEFING A CERTAIN RESCUE ACTIVITY. TO DISTINGUISH THESE POSSIBLES WE ALSO DID OTHER EXPERIMENTS TO SEE THE REVERSE RESCUE IN THE PARKINSON'S DISEASE FWE EVERYEXPRESS PINK 1 CAN IT RESCUE. OUR DATA SAID NO, THE ACROSS THE BOARD WE DID ALL THAT CONTROL, AND SO, IF THIS MODEL'S CORRECT, YOU KNOW IT MIGHT, BECAUSE IN MY PUSHING, PINK 1, BUT WE DID NOT SEE THE RESCUE. THE SECOND EXPERIMENT THAT IS ALSO VERY IMPORTANT IS TO DO A DOUBLE MUTANT. SO WHEN WE DID A DOUBLE MUSEUMITANT, WE SHOWED THE SAME PHENOTYPE AS REMOVING SINGLE GENE ALONE BECAUSE THIS WOULD BE COMPATIBLE WITH THIS 1, IF THIS MODEL'S CORRECT, 1 MIGHT SEE AN ENCHANTMENT OF THE PHENOTYPE WHICH IS--WHICH IS WHAT WE DON'T SEE. SO THIS, PLUS THAT THESE 2 PHENOTYPES IN OUR HAND SYSTEM IDENTICAL IN DROSOPHILA, SEVERAL ASSAYS, THE MOST PATHOMOANIC EXPLANATION IS THAT THESE 2 GENES FUNCTION IN THE COMMON GENETIC PATHWAY WITH PINK 1 REGULAR PARKIN AND REGULATES A DOWN STREAM TARGET THAT IS MIGHT MITOCHONDRIA, THIS IS IMPORTANT FOR SEVERAL REASONS, 1 REASON IS THIS REALLY DRAWS ATTENTION FOR MITOCHONDRIA DYSFUNCTION AS PART OF AT LEAST THE SUBSET OF PARKINSON'S DISEASE, A DISEASE MECHANISM AT THE CELLULAR LEVEL, ANOTHER THING IS JUST ACTUALLY OPENS UP A MORE SORT OF--IT'S INTRIGUING TO OPEN UP MORE INTERESTING QUESTIONS. ONE QUESTION IS WE KNOW THAT PINK 1 IS A KINASE AND WE KNOW THAT IS NO MITOCHONDRIA, WE FOUND THAT AS WELL, BUT PARKIN IS REALLY LARGELY CYTOSOLIC AND E3 LIGASE, SO THE QUESTION SHOW IS THAT--MOST PEOPLE DID NOT FIND AT THE TIME, THIS TO MITOCHONDRIA AND HOW IS IT ACTING THROUGH MIGHT MITOCHONDRIA THOSE ARE INTERESTING QUESTIONS, SO, BEFORE WE MOVE ON TO PAUSE TO ANSWER THIS QUESTION, THIS IS I GOT ALL THE TIME IN MY CLINIC. THE PATIENT ASKED: DOCTOR, YOU STUDIES SCIENCE AND ALZHEIMER PATIENT HYMER'S DISEASE, GREAT, DO IT FAST BUT YOU'RE WORKING ON WHAT? FRUIT FLIES AND WHY IN SO IN THE BACK OF THEIR MIND, THE QUESTION IS ARE THEY REALLY RELEVANT? SO WE THINK THIS PATHWAY IS RELEVANT BECAUSE A LOT OFO[iŤ KEY BIOLOGICAL PROCESSES ARE CONSERVED FOR HUMAN DROSOPHILA AND IT'S SPECIFIC TO THIS--TO THIS PROBLEM, WE COULD PUT HUMAN PINK 1 IN PARKINS LIKE THE GENES YOU AND I HAVE, WE PUT THAT IN FLYS AND RESCUES THEM, SO THIS IS GREAT BECAUSE THAT'S SUGGESTS THEY ARE NOT ONLY SEQUENCE HOMOLOGY, BUT ALSO FUNCTIONALLY CONSERVED SO AT THAT TIME, THEN THERE'S INDEPENDENT GROUP, COMPLETELY INDEPENDENT THAT WE DIDN'T SERE INFORMATION PRIOR, THEY JUST LOOKED AT CLINICAL FEATURES AND CONCLUDED THEIR CLINICALLY DISTINGUISHABLE, THERE'S A HUGE AMOUNT OF LITERATURE LATELY THAT LOOKING AT PINK PARKIN AND THEN THEY SHOW, LIKE SUBTLE MITOCHONDRIAL PHENOTYPE IN THE SELL CELL, I TRIED TO LIST IT AND I DID A REVIEW AND EVERYTHING IS IN THAT. I WROTE TOO MUCH. ALTHOUGH FOR MOST GROUP FIST YOU KNOCK OUT, YOU DON'T SEE A STRIKING PHENOTYPE, AT THE LEVEL OF DOPAMINERGIC ALONE WITH THE FEW EXPRESSIONS WE CAN ELABORATE LATER. AND THEY ARE SUBTLE CHANGES, BUT NOTHING AS STRIKING AS DROSOPHILA SOVICILLA, THIS IS CONSERVED MARKED FOR SOME OF THE WORK IS DONE HERE IN THIS PLACE. SO HAVING ESTABLISHED THESE 2 PROTEIN FUNCTIONS TOGETHER, THE NEXT QUESTION IS WHICH ASPECT OF MITOCHONDRIA PATHWAY FUNCTION DO THEY REGULATE, SO, WE--WE--YOU KNOW LATER I'LL TELL YOU WHAT PROMPTED TO LOOK AT THIS PROCESS CALLED MITOCHONDRIA DYNAMICS, BUT JUST TO SAY THAT WHEN I MOVED INTO THE FIELD, I WAS VERY NAIVE, MITOCHONDRIA, LIKE A BEAN SHAPED THING, SIT NOTHING THE DARKNESS OF THE CELL AND GIVE OUT ATP AND CREATE ALL THESE COMP LIT CATTED METABOLIC PATHWAYS I'M SUPPOSED TO REMEMBER, BUT CAN'T. BUT IT TURNS OUT THAT LATEST FINDINGS ABOUT THESE ARE ACTUALLY DYNAMIC, MEANING MOVING AROUND AND CHANGING SHAPES, YOU COULD MITOCHONDRIA LABEL WITH GFP AND RFP, THAT THEY'RE YELLOW, THIS IS USING THE CONTENT. SO THIS PROCESS IS MEDIATED BY MFN AND CONVERSELY, MIGHT O CHOND RAIA GENERATING 2 MITOCHONDRIA IN THIS PROCESS IS MEDIATED BY A PROTEIN CALLED GRP 1 AND BY THE WAY, HERE, MANY PEOPLE ARE ALSO LOOKING AT THIS PROCESS, I WANT TO TELL YOU THAT THE VERY FIRST GENE IN ALL ANIMALS THAT MEDIATES MITOCHONDRIA FUSION WAS IDENTIFY INDEED DROSOPHILA. SO WE WERE ASKING THE QUESTION, WHAT DOES THIS FIELD DIFFUSION PROTEIN DO IN DROSOPHILA BY LOOKING AT SAMPLE AGAIN SO THIS IS JUST A MUSCLE THAT WE DO A CONFOCAL BACK AND LOOK AT THE GREEN IS GFP AND RED IS LABELING MUSCLE SO WE CAN SEE THIS MIGHT O CHOND RAIA--MIGHT O CHOND RAIA IN BETWEEN SO WHEN WE REDUCE THE FUSION PROTEIN, YOU SEE THIS SMALL AND NUMEROUS AND AND THIS IS THE DRP IS SMALL FRAGMENTED. SO WHAT HAPPEN SYSTEM MITOCHONDRIA MORPHOLOGYST MAINTAINED BY THESE ROUGH ATOM SEASESS, WHY DO WE CARE ABOUT THIS? THIS IS WHAT I WILL SHOW YOU. SO THIS IS IN A WILD-TYPE MUSCLE, SO THIS IS AGAIN STAINING, LOOKING AT TISSUE INTEGRITY, THIS IS THE NORMAL. AND THEN THIS ALSO A MUSCLE SECTIONS BUT AS COMPARE TODAY PREVIOUS SCAN, THIS IS JUST A LOWER MAGNIFICATION AND THEN YOU SEE, THE MITOCHONDRIAL, TUNNEL NEGATIVE. PINK 1 MUTANT AS I SHOWED YOU, WE HAVE THESE AND MITOCHONDRIAL WHEN YOU LABEL THESE, THIS REDUCED THIS IS PROBABLY BECAUSE OF IMPORTING ERROR AND THEN THE TUNNEL IS STRONGLY POSITIVE. REMARKABLY WHEN WE EITHER, THE PINK 1 MUTANT BACKGROUND REDUCING THE FUSION OR INCREASE THE EXCISION THAT IT GOES THE SAME DIRECTION. SEVERAL PHENOTYPES ARE RESCUED, TUNNEL PHENOTYPE RESCUED, TISSUE PHENOTYPE IS RESCUED AND THE ANIMALS FLY BETTER LOCOMOTION IS BETTER SO THIS RESCUE IS COMPLETE. I PUT THE SLIDES OS BECAUSE IT'S NOT LIKE WE'RE IGNORING DOPAMINERGIC NEURONS, BUT WHAT WHEN WE LOOK AT THIS, IT'S MINIMAL. YOU COULD ACTUALLY BY LOOKING ATIT MITOCHONDRIA MORPHOLOGY, THIS IS A GROPE OF DOPAMINERGIC NEURONS THERE ARE SORT OF HAVING THESE CLUSTERS AND DRT PROTEINS CAN RESCUE THAT AS L. SO WITHOUT GOING INTO DETAIL, I WANT TO TELL YOU THAT LOTS OF PARKINS INDUCED SIMILAR IN THE PHENOTYPE WHICH CAN BE SUPPRESSED BY LOSS OF FUSION AND GAIN OF FISSION. SO THE NEXT QUESTION IS HOW DOES THE PINK PARKIN INTERACT WITH THE MACHINERY? SO WHAT WE DID IS LOOK AT THE DRP AND THE MSN AND FOUND THAT OVEREXPRESSION OF MIGHT O FUSEIN SHOWS THE SAME PHONOTYPE OF PINK PARKIN SO AGAIN THIS IS A MUSCLE SECTIONING, LOWER MAGNIFICATION AND PINK 1 SHOWS ALL THESE CLUMPS AS WELL AS OVERALL DECREASED LEVELS AND IT'S THE SAME, WE ALSO FOUND THAT BOTH OF THEM SHOW TUNNEL POSITIVITY; SO THIS IS JUST TO LOOK AT BECAUSE PARKINS IS A UBIQUITIN E31 LIGASE AND MSN IS THE PROTEIN WE COULD LOOK AT WITH A TARGET. IT TURNS OUT TO BE THE CASE, SO THIS IS JUST TAKING--I'M SORRY, THE DROSOPHILA IN THE FENCE, SO IN THE WILD-TYPE HAVE YOU A LOW LEVEL AND THE PINK PARKIN IS A DRAMATIC INCREASE IN THE LEVEL OF SO THIS ACTUALLY PINK PARKIN CAN BE--MSN CAN BE UBIQUITINNATED AND THEN YOU KNOW PINK PARKIN MEDIATED WITH THE DEGRADATION PROCESS THIS, IS ALSO JUST WANT TO ACKNOWLEDGE OTHER COLLEAGUES OTHER DONE BY OTHER GROUPS AS WELL AS MANY, MANY MAMMALIAN SYSTEM GROUPS INCLUDING RICHARD, AND MARK AND THEY'VE HAD MANY, MANY MANY OTHERS, SO THIS IS A COMMON ASSAY. SO WHAT IS THE OVERALL VIEW? IN IS A PATHWAY WITH AT LEAST IN PART MEDIA DEGRADATION MFN, AND THEN THIS PROCESS ALSO SHARE IN MAMMALIAN SYSTEMS AND THEN OR POSITIVELY REGULATING DRON P, SO IN THE PINK IN THE PATHOMITOCHONDRIA, REACHING A STATE THIS IS OVERFUSED FOR SOME REASON, AND THEN THAT HAS SOME PATHOLOGY AND THEN IF WE COULD HELP THEM BY REDUCING THEIR FUSION AND INCREASED SUFFICIENCY, THE WHOLE PHENOTYPE GETS RESCUED. LET ME JUST SUMMARIZE THIS PART, SO I TOLD THAT YOU THAT CAN ACT IN THE COMMON PATH WAY TO REGULATE THE INTEGRITY AND HUMAN AND DROSOPHILA PINK PARK IN THE THIRD. BUT THIS IS ACTUALLY VERY EXCITING, YOU COULD MODULATE MODIFICATION OF DYNAMICS WOULD SUPPRESS PARKINSON'S DISEASE, AND SO THIS REALLY SERVES A GREAT THERAPEUTIC STRATEGY, SO YOU DO SORT OF CELL BASED THERAPY RATHER THAN, YOU KNOW JUST FOCUS ON DOPAMINERGIC NEURONS. OKAY. I'M GOING TO PAUSE AND TELL YOU ANOTHER VERY EXCITING ASPECT OF THE PINK PARKINSON'S STORY MAINLY STARTED HERE AT NIH BY A VERY TALENTED RESEARCHER, RICHARD YOU'LL. SO HE HAS PUBLISHED SEVERAL DATA AND THIS IS COMING OUT FROM A REVIEW THAT HE WROTE. LET ME JUST SUMMARIZE SEVERAL OF HIS PAPERS AND JUST TO SAY THIS IS ALSO IN COLLABORATION BEFORE WITH MARK COOK SON AND A BUNCH--MANY PEOPLE AND MANY, MANY, YOU KNOW OTHER LABS ALSO CONTRIBUTED TO SURGEON AND COLUMBIA AND THIS--TOO MANY TO LIST, IT'S A EASY ASSAY TO DO, WHAT IT IS, IS HE HAS THESE HELIXES AND THEN HE TREATED WITH CCCP WHICH IS UNCOUPLER, WHAT DOES THAT MEAN? IT UNCOUPLES IN THE MITOCHONDRIA, WITH THE MEMBRANE WHICH IS CRUCIAL TO ALLOW TO YOU GENERATE ATP. SO WHEN YOU UNCOUPLE THESE MITOCHONDRIA, THIS MEMBER POTENTIAL WILL BE DESPATED AND THIS IS A SURROGATE MARKER FOR DAMAGED MITOCHONDRIA AND SO IN THIS KIND OF SITUATION, PINK 1 WHICH IS NORMALLY GOES TO MITOCHONDRIA, IT WOULD BE DEGREATED BUT WHEN THERE'S A DAMAGE, PINK 1 WOULD BE SPECIFICALLY STABILIZED ON THAT MITOCHONDRIA AND THAT PROCESS THEN RECRUIT PARKINS BUT SPECIFICALLY LOCALIZE TO THE ESTABLISHED MITOCHONDRIA AND THEN THIS SUBSEQUENTLY, LEADS TO UBIQUITINNATION TO SUBSTRATE INCLUDING MSF AND LEADS TO DEGRADATION OF THE PATHWAY, AND IN OTHER WORDS, THIS PINK PARKIN IS SITTING IN THE CROSS ROAD BETWEEN 1 IS UNDERSTANDING PARKINSON'S DISEASE, ANOTHER IS POTENTIALLY UNDERSTANDING MITOCHONDRIA QUALITY CONTROL BECAUSE BEFORE, I MEAN THERE WAS ALWAYS ARGUMENT WHETHER THERE WAS A SPECIFIC MORPHOLOGY ONLY TARGETING MITOCHONDRIA AT ALL, THIS ASSAY SAYS, YES INDEED AND, THE MOST IMPORTANT THING IS PINK PARKIN COME--CONFERS THE SPECIFICITY AND THIS IS EXCITING AND THEN THIS IS ALL GREAT, IT HAPPENS IN THIS CELL BRACED ASSAY--CELL BASED ASSAY SYSTEMS. SO THIS IS TO ME, WELL KNOWN THAT THE HYPOTHESIS IS PINK PARKIN PROMOTES REMOVAL, DAMAGE THROUGH AH TO HAVAGEY, AND THIS IS SPECULATION FROM MAYBE PARKINSON'S IS A FAILURE OF MIGHT O CHOND RAIA QUALITY CONTROL. THIS IS INTERESTING BUT I WANTED TO TELL YOU FROM OUR PERSPECTIVE, WE JUST WANT TO--WE WANT TO DRAW ATTENTION THAT THERE'S REALLY MORE TO PINK PARKIN AND THE MIRROR IMAGE TORO GBA FREE RADICALS THE STAND POINT, SO I'M GOING TO GIVE YOU TO VIGNETTES TODAY, FIRST IS ABOUT DEVELOPMENTAL ROLE OF PARKIN'S PATAY AND SECOND ABOUT TISSUE HEALTH. WELL, FIRST OF ALL DEVELOPMENT, SO THERE IS A STAGE DURING DEVELOPMENT, IN DROSOPHILA, AND IT'S A SPERMAT O GENESIS, WHY IS THAT? BECAUSE THIS IS INITIALLY WE FOUND THAT PINK PARKIN MUTANTS ARE ACTUALLY STERILE. SO WHEN WE LOOK AT THE PROCESS MORE CAREFULLY DURING THE SPECTROGENESIS, MITOCHONDRIA COME IN MANY DIFFERENT SHADES AND FORM, AND THEN THEY UNDERGO MITOCHONDRIA UNDERGO SELF-DYNAMIC CHANGES SO 1 STAGE LET ME JUST DRAW YOUR ATTENTION TO THIS IS WHAT WE CALL ONION STAGE. YOU HAVE REALLY LOVELY PAIRING AFRONs BLACK BALL AND A WHITE BALL. SO THE WHITE BALL IS THE NUCLEUS AND BLACK BALL IS A GERMAN WORD FOR MIGHT O CHOND RIATHEY FUSE TOGETHER, FORM THANKSGIVING ROCKETTURE. THIS IS THE SESSION, AND THEN, AS A RESULT OF 2 FUSED MITOCHONDRIA AND THEN THEY ARE THIS IS LGE ENOUGH, AND THIS IS UNDER THE MICROSTOP. WITH WHEN WE LOOK AT THE STAGE, SO THIS IS PINK 1 AND THIS IS WILD-TYPE, WE JUST GOT A HIGHER MAG NIHICATION, IN THEA KILLED WILD-TYPE YOU SEE THESE 2 SPHEREICAL STRUCTURES WITH THE HOMOGEANIOUS, WITH THE PINK 1 MUTANT, YOU SEE THE EVACUATION OF THE PROCESS. SO HERE'S JUST THE OVERALL VIEW. I WANT TO DRAW YOUR ATTENTION THAT THIS IS NOT BROKEN, YOU KNOW SOMEHOW LOSS OF CELL WALL, THIS IS JUST A PERFORATION ARTIFACT WHEN YOU CRUSH IT, IT'S JUST--SOMETIMES YOU CRUSH IT LIKE THIS, SOMETIMES YOU CRUSH IT LIKE THAT. BUT YOU SEE A LOT OF MIGHT ROUGH ATOM CHONDRIAL DEFECTS. SIMILAR THINGS HAPPENS IN PARKIN AND THEN ALSO WE COULD AGAIN SHOW THAT THIS PROCESS IS ALSO DEPENDENT ON PARKINS BY SHOWING THAT OVEREXPRESSION OF PARKINS. IN PINK 1 CAN RESCUE THE PHENOTYPE. SO AND RECALL TAKEN--THEY PINK 1 MUTANT IS 100% STERILE, WE COULD ACCOMPLISH SOMETHING LIKE 50% FEATERILE. AND AND THEN WE CAN PUT HUMAN GENES IN AND THEN THE STRUCTURES ALSO RESCUED. SO THIS IS 1 STAGE I MENTIONED TO YOU CALLED ONION STAGE BUT ALSO ENERGY STAGE THAT FOLLOWED THE ONION STAGE IN THAT THE [INDISCERNIBLE] WHICH IS MITOCHONDRIA STRUCTURE UNFURRAL AND LATER FORMING 2 LEAF BLADE STRUCTURE, SO THIS IS ACTUALLY INITIALLY PROMPTED US TO COME UP WITH THE DYNAMICS IDEAS WHEN WE LOOK AT THE FENEE TYPE WE SEE IN WILD-TYPE WE SEE 2 LEAVE PLATES WHERE IN PINK 1 AND PARKIN MUSEUMITANTS WE ONLY SEE 1. AND THEN THIS PHENOTYPE CAN BE RESCUED BY GENERATED ORDER OF ORDER OF MICRONS--GENOMIC RESCUED BACKGROUND. SO THIS SUGGESTS OF COURSE TO PROMPT US TO LOOK AT DYNAMIC ROLE, AND SUGGEST THANKSGIVING IS A DEFECT IN MITOCHONDRIAL MORPH O GENESIS, SO THEN WHAT ABOUT OTHER DYNAMIC MOLECULE, WE WERE DOING A GENETIC SCREEN FOR SUPPRESSION OF PINK 1 STERILITY AND HAPPENED TO IDENTIFY IT, A VIABLE MFN. AND IT IS IN GENERAL LETHAL, MOST REMARKABLY IF THIS TURNS OUT TO BE A DROSOPHILA HOM LOG OF A DISEASE MUTANT, 2, SO THIS ALLELE, NOT ONLY IS MFN ALLELE, BUT IT'S ALSO A DROSOPHILA DISEASE ALLELE THAT WHAT IS NUMBER 2, SHARKA MARIA 2 HAS NOTHING TO DO WITH YOUR TEETH, IT IS A GROUP OF DISEASE AFFILIATING PERIPHERAL NERVE. SO YOU CAN SEE THESE PATIENTS DURING THEIR TEENAGER YEARS, CLASSIC CASES THEY BECAME, THEY DEVELOPED FOOT DROP, THEY CAN'T WALK, HAMMER TOED AND HIGH ARCH, AND THEN, SO THEY HAVE ACTUALLY THESE DISTAL AX ORDER OF MICRONSS BECAUSE THEY'RE VERY FAR AWAY, LENGTH DEPENDENT DEGENERATE. THIS IS A VERY, VERY, COMMON--COMMON DISEASE AND MUTATIONS AND MFN 2, LEADS TO ACTUALLY THE MOST COMMON FORMS IN NEUROPATHY. WE HAVEN'T FOUND THE DROSOPHILA,A LEGAL OF IT SO WE WANT TO TELL THAT YOU THIS ALLELE SUPPRESSIONS MUSCLE DEGENERATION AND THIS IS AGAIN THE SAME ASSAY HERE JUST IN WILD-TYPE. THIS IS THE MUTANT BY ITSELF AND THIS IS PINK AND THIS IS PARKIN AND THEY HAVE THESE TUNNEL POSITIVITY BUT IN THE BACKGROUND, THERE'S AN ALLELE THAT IS SUPPRESSED BUT OTHER EXPERIMENTS WE'RE FINDING SUGGESTING THAT THE ALLELE BEHAVEs AS A PARTIAL MORPH AND PARTIAL LOSS OF MFN. SO THE IMPORTANT THING IS WHAT HAPPENS IN TESTES, SO THIS ALSO SUPPRESSS MALE FERTILITY AS WELL AS MITOCHONDRIA MORPH LOGICAL DEFECTS AT THE ONION STAGE AS I MENTIONED DUE TO BEFORE, DUE TO PINK 1 HERE, JUST TELLING YOU THE STRIKING SUPPRESSION OF MALE TERILLEGALSITY. SO IN PINK 1 MUTANT WE SEE 0 FEATERILE WHICH WE TAKE A SINGLE FLY TO LOOK AT IN THE PINK 1 YOU SEE A HUNDRED FOR THAT FERTILITY. SO WHAT DOES THAT MEAN? WHAT I WAS TRYING TO TELL YOU AS A FIRST VIGNETTE IS THE PINK MIGHT HAVE USED THEM AND THEY REGULATE A DEVELOPMENTAL MORPH LOGICAL CHANGES REQUIRED TO DEVELOP FUNCTIONAL SPERM IF ANYTHING THIS IS PROBLEMATIC THEY DON'T--THEY'RE STERILE. SO THE MALE PHENOTYPE DUE TO PINK PARKIN MSN AND UNLIKELY DUE TO MIRROR IMAGE TO HAVAGEY, I WAS THINKING THERE ARE MORE TO THIS THAN JUST MIGHT O--YOU KNOW THE MITOPHAGEY, SO WHAT I WANT TO TELL YOU IS COMING BACK TO THIS RICHAR YOU'LL'S THEME HERE OF MIRROR IMAGE TO HAVAGEY, I WANT TO TELL YOU THAT MITOPHAGEY UTELLIZES AT LEAST IN PART THE MACHINERY SO HERE AFTER THE PROTEIN IS UBIQUITINNATED AND IT'S RECRUITED THEN GRADUALLY THE MITOCHONDRIA IS ELIMINATED, SO, A LOT OF PEOPLE HERE STUDYING AUTOPHAGEY, BUT LET ME GIVE YOU A SIMPLIFIED VERSION OF INTRODUCTION, SO AUTO TO MY KNOWLEDGAGEY IS UNWANTED CELLULAR CONTENT TO ORGANELLES OR ELIMINATED BY VARIOUS STRESS OR STARVATION, AMONG OTHERS, SO THIS IS THE PROCESS OF OF ASSEMBLIER AND LEADING TO DAG GRADATION BY LIAISON STUDIES OF MULTIPLE ENDOCRINE, SO THERE ARE 2 KEY GENES INVOLVED, AND 1 IS CALLED ATT 1 AND INVOLVES INDUCTION OF PHAGOSOME AND THE OTHER IS ATT7 LEADING TO THE PROCESS OF MITOPHAGEY. SO THEN SINCE WE PREVIOUSLY FOUND THIS DYNAMICS ANGLE, SO WE ASKED WHAT IS REALLY THE RELATIONSHIP BETWEEN MITOCHONDRIA DYNAMICS AND MITOPHAGEY WITH MAINTAINING TISSUE HEALTH IN PINK PARKINS MUTANTS. SO THIS PAPER SAYS IT ALL, WHAT THEY DID IS THEY LOOK AT THE MORPHOLOGY AND FOLLOWING THE FETE OF THE MIGHT O--FEAT OF FETE OF--FATE OF THE MITOCHONDRIA, UNDERGOING A PROLONGED PHASE CALLED SOLITARY PERIOD AND ONLY DURINGAVE SHORT PERIOD OF TIME THEY FUSE. AND THEN, THIS SORT OF A NETWORK STAGE AND FOLLOWING IN THAT, THE UNDERGOING FISSION, AND THEN FOLLOWING FISSION, THESE MITOCHONDRIA HAS 2 DIFFERENT PHASES SO 1 IS HAVING AN INTACT MEMBER POTENTIAL AND ANOTHER GROUP, ACTUALLY HAS DEPOLARIZED MEMBER POTENTIAL AND THEN WHEN THEY APPROXIMATELY THIS DEPOLARIZED MEMBER POTENTIAL, THEY FOUND 2 THINGS, 1 THING IS, SOME OF THEM RECOVER AND GO BACK TO THE CYCLE, SOME OF THEM THEY CONTINUE LOSING THE POTENTIAL, THEY UNDER GO AH PHAGEY. SO--AUTOPHAGY. SO THE IMPLICATION IS ALSO NEVER REALLY FORMERLY STATED. IT'S FISSION CREATES A SUBSTRATE FOR MITOPHAGEY, A SMALL AMOUNT OF FRAGMENT THAT KOOND CANNOT REFUSE WITH THE NETWORK. SO WHY DOES INCREASE FISSION RESCUE THE PHENOTYPE AND IS IT BECAUSE IT IF TAILITYS THE MITOPHAGEY PROCESS AND IF SO, THE PREDICTION WOULD BE OVERSUPPRESSION WOULD NO LONGER SUPPRESS IT AND MUTANT, RIGHT? ON HOWEVER THIS IS NOT ONLY WHAT WE SEE, SO WHEN WE SEE IS WHEN WE OVEREXPRESS, THIS IS PINK 1, PINK 1 WITH OVEREXPRESSION, THIS IS JUST THE LOWER MAGNIFICATION AND HIGHER MAGNIFICATION AND THIS RESCUE STILL OCCURS IN FLIES LACKING ATG 7 AND THEN ALSO IF YOU LOOK AT IT FURTHER THIS STILL OCCURS IN ATG 1 MUTANTS: SO IN OTHER WORDS, WELL WE'RE SAYING IS AUTO PHAGEY, THEY CAN RESCUE ANY ABSENCE OF THESE AUTOPHAGY MUTANTS, IN THE PRESENCE OR ABSENCE OF THESE MUTANTS. SO THERE'S ALSO THIS INTRINSIC INTRIGUING OBSERVATION IF YOU JUST DO A DOUBLE KNOCK OUT OF MFN 1 AND 2, SOk8 Ť MITOPHAGEY IS OCCURRING SO AT LEAST IT'S DISPENSABLE FOR IT, SO OUR SPECULATION 3—er RESILIENCE CONCILE THESE PROCESSES IS THAT IF YOU JUST WANT TO RESTORE--I MEAN RESTORE THE BALANCE BETWEEN FUSION AND FISSION, IT'S SUFFICIENT TO RESCUE MITOCHONDRIA FUNCTION AND TISSUE HEALTH INDEPENDENT OF AUTOPHAGY OR MYTO HAVAGEY, OF COURSE ANOTHER INNERTERPRETATION MAY BE DRP OVEREXPRESSION BASE STIMULATE QUALITY CONTROL THROUGH AGG 1 AND 7 INDEPENDENT PATHWAY. THIS IS KIND OF INTERESTING TO EXPLORE FURTHER. SO JUST SORT OF THINKING ABOUT PARKINS PATHWAY IS THAT THIS PATHWAY REGULATE MIGHT O CHOND RIFUNCTION AND TISSUE--MITT O CHONDRIAL FUNCTION AND MORPHOLOGY. WE LOOK AT THAT IN VIVO, AND MOST RECENTLY THEY'RE ALSO WORK DONE IN THE PATHWAY ALSO FUNCTION IN THE MITOCHONDRIA MOVEMENT SO. SO IT WILL BE INTERESTING AS THE FIELD MOVES ON, TO SEE WHAT'S REALLY THE CORE, WHAT IS REALLY THE PROBLEM, THAT APPROXIMATE PINK PARKIN MUTANT IS BRINGING B. OKAY. SINCE I HAVE A LITTLE BIT MORE TIME I'M GOING TO TELL YOU ANOTHER--SWITCH GEARS TO TELL YOU ANOTHER STUDY DONE IN MY LAB. LOOKING AT ANOTHER GENE CALLED MARK 2. FOR THE MOSTIVATION IS ALSO IN OUR MINDS IS PARKINSON'S DISEASE 1 DISORDER OR A SYNDROME OF OF MANY DISEASES, BUT THE SAME FINAL OUTCOME. SO, IT IS HARD TORE ADDRESS THIS QUESTION, IN A HETEROGENEOUS POPULATION OF SPORADIC CASE, BUT IN A GENETIC FORMS WE COULD ATTEMPT THIS QUESTION, SO AS WE KNOW, THE PINK PARKIN IS AUTOSOMAL DOMINANT FORMS OF PARKINSON'S DISEASE, WHEREAS LRRK 2, LEADS TO ROOT STUDIES OF MULTIPLE ENDOCRINAL DOMINANCE, DID I SAY--PICK PARKIN'S RECESSIVE LRRK, IS AUTOSOMAL DOMINANT. I WANT MAKE SURE I'M NOT MIXING IT UP. AND AN EXCITING THING AND IT ALSO WORK INDEED SEVERAL LABS AND HIGH AND MARK COOKS MAN AND MAYBE MORE, SO, THAT IT IS REALLY THE SINGLE MOST COMMON GENETIC CAUSES, NOT ONLY AS A HIGHER PREVALENCE IN GENETIC, BUT CERTAIN GENETIC POPULATIONS, THE CARRIER RATE CAN BE AS HIGH AS THE 40% AND ANDIE SINGLETON DID THE GWAS STUDY TOGETHER WITH SOME OTHER PEOPLE IN THE FIELD SUGGESTING MUTATIONS IN LRRK 2, ASK ALSO A RISK FACTOR, IN SPORADIC CASES AND THIS IS VERY INTERESTING. THE COMPLICATION IS, THIS GENE IS IMPLICATED IN JUST ABOUT EVERY CELLULAR COMPARTMENT AND EVERY PROCESSES, AND IT'S ENDOGENOUS--IT'S EXPRESSION HAS BEEN IMPLICATED IN JUST ABOUT EVERY FELLOW COMPARTMENT SO IT'S HARDER WHEN YOU SUBMIT A PAPER, YOU SAID IT'S ENOUGH COMPARTMENT, YOU HAVE NO NOVELTYY BECAUSE SOME PEOPLE ALREADY MENTION IT BUT IT'S CONFUSING TO SEE EXACTLY WHERE IT IS THAT IT FUNCTIONS. SO, WHAT WE'VE STARTED OUT BY ASKING A SIMPLE QUESTION, DOES LARK 2 REGULATE INTEGRITY BECAUSE WE HAVE AN ASSAY, WE GENERATED A DROSOPHILA MUTANT OF THE MOST COMMON LRRK 2 MIN DUCE THE MUTATIONS, WHICH IS, GS MUTATION THIS IS IS JUST THE STUDY, SO WE LOOK AT MITOCHONDRIA, IT LOOKS GREAT AND MUSCLE, THERE'S NO PHENOTYPE AT ALL AND WE LOOK AT MITOCHONDRIAL MORPHOLOGY, LOOKS GREAT, SO, CAN'T REALLY SAY, THIS IS HAVING ANY PHENOTYPE, IN TERMS OF REGULATING MITOCHONDRIA INTEGRITY CROSS LINKING SO THEN WE LOOKED AT WHAT IS--WHAT DOES IT DO ENDOGENOUSLY SO IN DROSOPHILA WHEN WE AND WOO HAVE THIS PROTEIN AND REMOVE IT, IT HAS 2 PHENOTYPES 1 IS NEUROMUSCULAR JUNCTION PHENOTYPE AND ANOTHER IS A FEMALE PERFILLITY PHENOTYPE SO THESE FEMALES HARBORING THIS MUTATION AND REDUCED FERTILITY, AND THIS CAN BE SPECIFICALLY RESCUED BY LRRK EXPRESSION IN A GROUP OF CELLS CALLED FOLLICLE CELLS, NOW WHAT DOES FOLLICLE CELLS, DROSOPHILA FEMALE GERM LINE IS A PROGRESSIVE PROCESS AND THERE ARE GERM LINE CELLS AND NONGERM LINE CELLS, FELL COLCELLS AND NONGERM LINE CELLS IT'S MONOLAYERED EPITHELIAL CELLS COVERING THE CELLS AS THE GERM LINE AND THEN FOR THIS TOP PURPOSE, I WANT TO SHOW YOU THIS IS WHAT YOU ACTUALLY SEE, IS THE MONOLAYER EPITHELIAL CELLS, IS GREAT FOR CARRY OUT CELL BIOLOGY STUDY. SO WE'VE STARTED OUT BY ASKING THE QUESTION, WHERE EXACTLY IS IT EXPRESSED. IT'S NOT A TRIVIA QUESTION BECAUSE IT'S SYNTHICATED EVERYWHERE AND WE GENERATED TRANSGENE RESCUE TRANSGENE UNDER IN. DODGE NOWS CONTROL AND IMPACTED AND LOOKED AT IT SO THIS IS JUST 1 SUCH CELL IN THE FOLLICLE CELLS, AND THEN THE CIRCLE DELINEATES WHERE THE NUCLEI--NUCLEUSS SO THIS IS DOUBLE STAIN. THIS IS IMAGE SUBMERGE PROGRESS THE STAINING. SO WHAT WE SEE SEE IS WHEN WE SEE CLEARLY NO OVERLAP AT ALL. THIS IS INCONNIVITIENT WITH OUR THINKING IN IS NOT DUE TO MITOCHONDRIA INTEGRITY. SO WHERE IS IT EXPRESSED. WE FOUND IT'S EXPRESS INDEED THE LYSOSOME, SO WE KNOW--YOU KNOW, LOU HAN'S ALSO WORKING ON THIS PROCESS AND WHAT IT IS, AS YOU KNOW SO THIS IS EXTENSIVE FROM THE END O STUDIES OF MULTIPLE ENDOCRINES TO LYSOSOMES SO WHEN WE LOOK AT TWE FOUND THAT LRRK 2 AND EXPRESSED IN THIS AREA, AND THIS IS 1 OF THE THIS IS AGAIN A SINGLE CELL RESOLUTION, SO IT'S REALLY LIKE A CIRCLE ON THE MEMBRANE, AND WE LOOK AT IT CAREFULLY CAN T CAN CO LOCALIZE, ARE RAB 7, IT'S LOCALIZED TO LAMP 1 WHICH IS A MARKER;LyB+w AND LYSOSTUDIES OF MULTIPLE ENDOCRINE. SO WHAT IS THE PHENOTIME. WE FOUND THAT FOCUSING ON THIS OTHER MARKER, BUT HERE'S PANEL IS WILD-TYPE, TOP PAN CELL LARGE MUTANT. WHAT WE FOUND IS THAT THESE ARE AGAIN, THE AND ONLY IN LRRK, YOU HAVE THIS EXPANSION OF RAB 7 COMPARTMENT AND THEN,--THEN WE SAID WHAT DOES THAT MEAN. WHAT WE DID IS WE DID A POST HASTE EXPERIMENT, BASICALLY TAKING THESE EGG CHAMBERS OUT AND PUT A TRACER WHICH IS A DEXTRAN WHICH IS THE FOLLICLE CELLS AND THE ENDO SIGNIFYITOSEIN AND FOLLOW THE FATE AND STOP AFTER 10 MENNITES AND AFTER A WHILE THE TRACER WILL BE THE RAB 7 COMPARTMENT SO THIS IS LARGE LABELING. HOWEVER IN LRRK MUTANT WE SEE THE TRACER, SOMETIME WEES DON'T HAVE TRACERS, SOMETIMES YOU SEE THIS STUCK IN RAB 7 SUGGESTING A DEFECT IN THIS LATE ENDO STUDIES OF MULTIPLE ENDOCRINE. SO NEXT WE LOOK AT USING THE LYSOTRACKER TO LOOK AT WHERE THESE--TO LOOK AT WHERE THESE LYSOTRACKERS, LIES LOW STUDIES OF MULTIPLE ENDOCRINES--LIES SOPHISTICATED STUDIES OF MULTIPLE ENDOCRINE TRAVELED. SO THIS SHOWS ROUGH LOW THE SAME. WHERE IN RAB 7, WHEN WE DO CONSITTATIVELY RAB 7 WE SEE MORE CLUSTERS IN THE NUCLEUS. THIS PHENOTYPE CAN BE MODIFIED WHEN WE--SO THIS IS RAB 7, WHEN WE LOSE A FUNCTION OF LRRK 2, THERE'S SIGNIFICANT INCREASE, THERE'S SIGNIFICANTLY SUPPRESSED SUGGESTING THAT LRRK CAN MODULATE RAB 7 INDUCED CLUSTERING, SO THIS ROUGH ATOM SEASES ALSO IN THE LRRK, MUTANT OCCURS WITH THIS LYSOSOMAL CLUSTERING. AND YOU KNOW WE ASK QUESTIONS ABOUT HOW IS LRRK 2 AFFECTING RAB 7, WE LOOK AT THIS CO IP STUDY AND THEN FOUND THAT LRRK CAN SPECIFICALLY INTERACT WITH RAB 7 AND JUST BY THE WAY, WE ALSO FOUND IT CAN PHYSICALLY HAVE 5 AND IN A SMALL PAPER IT'S PUBLISHED, BUT THIS RAB 7 IS WHAT WE ACTUALLY CAN SEE, THE PHENOTYPE OF, SO IN A WAY, BECAUSE THE TIME CONSTRAINTS, I JUST WANT TO SUMMARIZE THIS FINDING, SO, LRRK, WE FOUND--LOCALIZED TO THE MEMBRANE DODGE NOWS PART, THE LOCI OF THE MEMBRANE APPROXIMATE PART, THE HIGHLIGHTED PIPE LRRK, IS ABLE TO REGULATE OR MODULATE REFRESH YOUR RECOLLECTION AB7'S ABILITY TO PROMOTE LIAISON STUDIES OF MULTIPLE ENDOCRINAL CLUSTERING. AND THE MUTANT BEHAVES HOP SIT TO LRK WILD-TYPE. AND BIND TO?– 7y LRRK, 7 SO I DIDN'T GET THE CHANCE TO TALK TO YOU BUT WE CAN PUBLISH AND WE CAN GO ONLINE AND READ ABOUT THIS. SO WE GENERATED A SENSITIVE MODEL FOR HOW WE PUT TOGETHER THIS PROCESS. WHAT WE THINK IS THAT THIS LYSOMOL, THIS 7, ACTUALLY THROUGH SOME BINDING PARTNERS REALLY INTERACT WITH DINING BY 19 COMPLEX AND THEN MICROTUBE Y'ALL, SO IN THE NORMAL SITUATION, LRRK, POTENTIALLY INHIBITING OR MODULATING THIS PROCESS. WHEREAS IN THE--IT BINDS TO RAB 7 BUT INHIBITING THIS PROCESS, IT IS POSSIBLE TO SIT THROUGH THE BINDING THE GDP FORM BUT IN THE BINDING WE FIND THE WILD-TYPE STRONGER THAN GDP FORMS. RAB AND HOWEVER, WHEN LRK IS MUTATED, WE THINK RATHER THAN SUPPRESSING THIS PROCESS, IT ACTUALLY PROPROMOTES THIS PROCESS. THAT INVOLVING MORE DYNAMIC MOVEMENT TOWARDS THE MINUS END AND MICROTUBIAL THAT'S WHY IT'S CLUSTERED IN THE LYSOSTUDIES OF MULTIPLE ENDOCRINES, SO THIS MODEL IS CONSISTENT WITH SEVERAL FINDINGS IN THE FIELDS SUGGESTING THAT IN THE LRRK MUTANT IT HAS NEURONAL PROCESSING DEFECT, YOU KNOW IN DENDRITES AS THAT THAT'S PARTIALLY RELATED TO MICROTUBIAL AS WELL AND THEN OF COURSE MORE NEEDS TO BE TESTED BUT THIS IS A START TO ALLOW US TO THINK ABOUT THIS PROCESS. HOW HOW DO WE PUT THIS TOGETHER? SO ALTOGETHER, IT SEEMS TO BE THAT FAMILIESIAL PARKINSON'S DISEASE INVOLVING MULTIPLE PATHWAYS, 1 IS THE PING PATHWAY I TOLD YOU ABOUT LEADING TO MITOCHONDRIA DEFECT, THERE IS ANOTHER GENE, AND MARK HAS DONE A LOT OF WORK ON IT, IT INVOLVES IN ASPECT OF THE STRESS HAD LINKAGE TO PINK PARKIN BUT NOT REALLY BUT NOT DIRECTLY RELATED TO PINK PARKIN SO WE THINK IT'S PARALLEL AND I TOLD THAT YOU LRRK, 2, WE THINK IT'S REGULATED PROCESS OF ENDOLIAISON STUDIES OF MULTIPLE ENDOCRINAL PATHWAY AND THEN DATA HERE, ALSO FROM NIH SUGGEST THANKSGIVING'S MAYBE INTERACTIONS BETWEEN THIS AND LRK 2 AND SO REALLY MORE THAN 1 PATHWAY INVOLVED IN CLOSING PD AND IN THE FUTURE CALLS FOR PATHWAY SPECIFIC THERAPY SO YOU CAN OPTIMIZE YOUR TREATMENT POTENTIAL MINIMIZE SIDE EFFECTS. I JUST WANT TO ALSO DRAW YOUR ATTENTION BECAUSE I AM BY NO MEANS SUGGEST THANKSGIVING IS NOT, THERE'S NO CROSS TALK, IT JUST OBVIOUSLY, SENDS MITOCHONDRIA WHICH UNDERGO MYTOPHAGEY, SO THE DEFECS AND LYSOSONES, LEAD TO DOWN STREAM LEVELS AT'RE ARE NOT AT INITIAL LEVELS SO THIS IS INTERESTING TO SEE. AS I'M A CLINICIAN, I'M AX CUTELY AWARE OF THE CLINICAL IMPLICATIONS OF OUR DROSOPHILA WORK. SO WHAT DOES THAT MEAN, SO LOOKING FROM DROSOPHILA WORK AND PROJECT TAG IN CLINICAL STUDY, WE CAN SEE THAT PARKINSON'S DISEASE POTENTIALLY, A GROUP OF MOLECULAR LE DIVERSE GROUP OF DISORDERS, SOME OF THEM AFFECTING MITOCHONDRIA AND SYSTEMS OF THE FUTURE, SO A PERFECT TREATMENT WOULD BE PATHWAY SPECIFIC. TREATMENT. SECOND OF ALL, MITOCHONDRIA DYNAMICS MAY BE A NOVEL TARGET FOR MITOCHONDRIAL FORMS OF PARKINSON'S DISEASE, IN DROSOPHILA, IT WORKED SO WELL, REGULATES CELLULAR AS WELL AS ORGANIZATIONS NIGHS MAL. THE PRESENCE OF THESE NEURONS JUST AS IN HUMAN PATIENTS THEY HAVE NONMOTTOR SYMPTOMS SO THERAPY IS TARGETING CELLULAR DRUGS ARE REALLY NEEDED TO EFFECTIVELY TREAT BOTH MOTOR AND NONMOTOR SYMPTOMS SO I JUST WANT TO ACKNOWLEDGE THE PEOPLE WHO DID THE WORK. THIS IS THE ORIGINAL GROUP WHO DID THE PINK PARKIN 1 EMPLOY IRA GOT A POSITION AT UCLA AND MSTP STUDENT AND DOG A RESIDENCY AND DETERMINED TO A PHYSICIAN SCIENTIST. TECHNICIAN GOING TO GRADUATE SCHOOL, THIS IS THE POST DOC FROM WHO HAS A POSITION IN CHINA, AND THEN AGAIN, MARK THIS IS THE WHO IS A POST DOC DID THE DYNAMICS WORK AND THEN, THAT GENE--I DIDN'T GET A CHANCE TO TALK ABOUT YET. AND THEN THIS IS CURRENT MEMBER IN THE LAB, THIS IS, PAST MEMBERS AND THESE ARE THE PEOPLE WHO PROVIDED WITH US REAGENTS AND THEN TEACHES US VARIOUS TECHNIQUES AND THESE ARE FUNDING SOURCES INCLUDING GENEROUS SUPPORT FROM NIH. SO THAT'S IT AND I'M READY FOR QUESTIONS. [ APPLAUSE ] >> [INAUDIBLE QUESTION FROM AUDIENCE ] >> I ACTUALLY HAVE A SLIDE LEER. SO WE LOOKED AT INTERACTION BETWEEN PERO QUAT AND RODENTS, THOSE ARE MIGHT CHONDRIAL TAXINS WORKED UP IN LATE 1980S SO THE AFFECTS EITHER INCRISEOXIDATIVE STRESS OR COMPLEX 1 OR A COMBINATION OF EFFECTS SO WE TAKE PIRVEG 1 FLIGHTS, AND X IS HOURS OF TREATMENT AND THEN WE FOLLOW THE TIME COURSE. BUT THE Y AXIS IS THE AMOUNT OF SURVIVAL. SO HERE THERE ARE 3 LINES, 1 LINE IS PINK 1, PINK COLOR, OKAY? THE OTHER 2, 1 IS„3wx ISOGINIZED WILD-TYPE, ANOTHER IS PINK 1 MUTANT WITH RESCUE CONSTRUCT, SO WE WANT TO BE VERY ANNAL ABOUT OTHER BACKGROUND MUTATION, SO WE SEE IT STARTING FROM THE BEGINNING THERE ARE A HUNDRED% VIABLE AND AS TIME GO ON, THESE FLIES ULTIMATELY WILL ALSO DIE TO KEEP THIS AS A HUGE TOXIN. SO PINK 1 THEY WILL DIE MUCH FASTER, THE RODENT DOES THE SAME THING. SO THE MUTANTS CAN ACTUALLY HAVE INCREASED SENSITIVITY, THE MUTANTS HAVE INCREASED SENSE TICHITY WITH THE STRESS. WHETHER OVER EXPRESSION CAN RESCUE? THAT EXPRESSION IS A GOOD 1. IT PROBABLY WILL BE MORE RELEVANT TO PARKIN BECAUSE WHEN WE OVEREXPRESS PINK 1 IF HAS TOXICITY, IT SHOWS THE DIFFERENT TISSUES PATHOLOGICAL EFFECTS THAT WE DIDN'T ACTUALLY DO THAT SO JUST REMEMBER, DOING THIS EXPERIMENT IT'S EASY TO SEE BUT TO DO IT RIGHT YOU NEED ALL THESE GENO TYPE TO BE CONTROLLED AND BACK CROSSES, IT'S NOT TRIVIAL EXPERIMENT TO DO IT RIGHT ALTHOUGH YOU SEE A LOT OF FINDINGS EVERYWHERE. >> DO YOU THINK THAT THESE MOLECULAR PATHWAYS COULD ALSO BE SIMILAR IN ALS WHICH IS ALSO--WHICH IS PART OF THE MOTOR NEURON DISORDER, I MEAN LIKE DO YOU EXPECT SIMILAR PATHWAYS, I MEAN LIKE A MIGHT O CHOND RIIAL DYNAMICS IN THE DISORDER SUGGEST ALS WHICH IS A NEURALONAL DISORDERED? >> THAT'S AN INTERESTING QUESTION BECAUSE ALS A NEURODEGENERATIVE DISEASE AND IT'S MAIN DEFECT, WELL ARE ALSO DIFFERENT GENETIC FORMS OF IT, THERE'S 1 FORM, SOD 1, AND IT HAS A STRONG MIGHT O CHUBBEDRIAL COMPONENT---MITOCHONDRIAL COMPONENT, SO THERE ARE STUDIES SUGGESTING MAYBE RELATED TO DYNAMICS. THE MEAN THE INTERESTING THING IS, WHETHER INCREASING TOPOLOGY CONTROL COULD RESCUE THE PHENOTYPE. YEAH, BUT LOOKING AT LRRK 2, THERE'S NO,--I MEAN PATIENTS DO NOT HAVE ALS. >> HOW ABOUT THE ROLE IN THE MITOCHONDRIAL TRANSPORT OR AXONAL TRANSPORT. >> THAT'S A WONDERFUL QUESTION, HAVE YOU A GREAT COLLEAGUE LEER, LOU HAN, CHEN WHO JUST PUBLISHED A PAPER IN CURRENT BIOLOGY SUGGESTING THAT ACTUALLY, AX ORDER OF MICRONS TRANSPORT, YOU KNOW THESE PINK PARKIN--TRANSPORT MIRROR IMAGE TO HAVAGEY TRANSPORT TO THESE DAMAGED MITOCHONDRIA CCP TREATED MITOCHONDRIA CAN BE DEGRADED. YOU GUY K'S--GUYS CAN TALK. RICHARD? >> [INAUDIBLE QUESTION FROM AUDIENCE ] >> RIGHT. >> RIGHT. >> IT'S A VERY INTERESTING QUESTION, SO, YOU KNOW I WOULD LOVE TO HAVE SOMEONE HAVE A SAMPLE TO DO THESE HUMAN STUDIES, BUT THE THING IS SOMETIME WHEN IS YOU HAVE DO DISEASES, BAD THING, BUT OCCASIONALLY IT'S A GOOD THING, RIGHT? OH, YEAH, YEAH, SO I THINK IT'S THE PINK PARKIN IS NOT THAT RARE, BUT IT'S RELL RELATIVELY RARE, THE QUESTION IS, YOU KNOW--WE DO HAVE SOME PEOPLE WHO ARE INTERESTED IN THE SHOCKA MAY AREA 2 THEN-- >> [INAUDIBLE QUESTION FROM AUDIENCE ] >> RIGHT. YES. --OKAY, GOOD AHEAD. YES. >> [INAUDIBLE QUESTION FROM AUDIENCE ] >> YEAH, IT IS INTERESTING WHEN WE LOOK AT, OH, OKAY, SO REPEAT THE QUESTION. OKAY. SO, THERE ARE SEVERAL ASPECT 1 IS ABOUT IF WE FOUND THE ALLELE THAT CAN SUPPRESS THE PINK 1, THE QUESTION IS IN HUMAN CASES WE DON'T KNOW BUT WE WOULD LOVE TO KNOW IF HAVE YOU A LOT OF PATES YOU CAN SCREEN, SOMETIMES YOU ENDED UP SO UNFORTUNATE TO HAVE 2 BAD DISEASE MAY NOT BE THE BAT THING, YOU END UP HAVING A GOOD OUTCOME, IT'S INTERESTING, THIS IS THE IMPLICATIONS, THIS IS THE SPARK, SO SECOND OF ALL, HE ALSO ASKING I SAID THAT MAYBE PINK PARKINS GOES BEYOND QUALITY CONTROL, AND THAT HE SAID YES BECAUSE IF YOU LOOK AT THE MITOCHONDRIA, IT'S NOT JUST THE MORPHOLOGICAL, IT'S BROKEN SIGNIFICANTLY LONG, WHAT IS GOING ON. SO IT IS INTERESTING, IF YOU LOOK AT WHEN WE LOOK AT MFN, ALLELES IT ALSO HAS THESE PRISTINE PHENOTYPES AS WELL BUT THE QUESTION IS WHY IN I DON'T KNOW. SO MFM N IS--MFN SHOULD BE OUTER MEMBRANE BUT THE MUTANT ALSO HAVE A PRIVATINE PHENOTYPE. >> RIGHT. >> WE HAVEN'T DONE THAT EXPERIMENT TO DO IT RIGHT, LIKE WE--YOU THEN IS VERY SIMPLE, BUT WE DO BACKGROUNDS AND ALL THESE CLEAN GENETICAC GROUND AND WE DID THAT, WE HAVEN'T DONE THAT EXPERIMENT. SO WE DON'T KNOW. AND ALSO, BEAR IN MIND, FUSION INFISSION MOLECULE WHEN IS THEY'RE EXPRESSED THEY'RE LETHAL SO IT IS DIFFERENT. THEN DO YOU TISSUE SPECIFIC KNOCK DOWN AND OVEREXPRESSION, SO PINK PARKIN IS VIABLE AND YOU CAN FOLLOW THE DRAIN ED? >> [INAUDIBLE QUESTION FROM AUDIENCE ] >> YES, YES, YES. SO LET ME GO BACK TO THIS SO PEOPLE CAN SEE THE SLIDE. THIS, YES. SO IT IS, ACTUALLY, I JUST DIDN'T GET THE CHANCE TO TALK ABOUT. SO, THIS GROUP, AND 2 A LESS EXTENT, DJ 1 IS WELL, THEY'RE ALL RECESSIVE AND THERE ARE TENDS TO BE JUVENILE ONSET AND SOMETIMES THEY START THEIR AGE WHEN THEY'RE 30, AND EARLIER BUT IT'S VERY UNFORTUNATE. BUT THERE ARE NONE THE LESS A HUNDRED% PENETRANT. SO LRRK, 2, IS CAN BE EARLY STAGE AND LATE STAGE. SO THEY'RE NOT A HUNDRED% PENETRANT. SO THERE IS ALSO A STRIKING PHENOTYPE AND PARKIN THERE'S NO LOWY BODY, THERE'S NO LUNG CANCERY BODY AND THEY'RE A PROTEIN AGGREGATE THAT SPECIFICALLY DOPAMINERGIC NEURONS, THAT'S POSSIBLY TO THE PATHOLOGICAL HALLMARKS SO IN THE NUCLEUS IS ALSO A COMPONENT IN LOUIS BODY BUT I WANT TO TELL YOU, LOUIS BODY KIN--CONTAINS A LOT OF DIFFERENT THINGS SO BY BEING IN LOUIS BODY DOESN'T MEAN MUCH, DOESN'T MEAN LIKE 1 PROTEIN SUPPORTED OR NOT, BUT NEVERTHELESS, IN PARKIN MUTANT FOR THE MOST STUDIES I MEAN THERE'S SEVERAL OUTLIERS THERE'S NO LOUIS BODY BUT THERE IS 1 STUDY, ONLY 1 PATIENT THIS FAR, UNFORTUNATELY PASSED AWAY AND THEN THE BRAIN WAS LOOKED AT. SO IN THAT SETTING, THERE IS LOUIS BODY, BUT THIS IS NM1, MORE DATA WOULD BE GREAT AND THEN DJ1 WE DON'T HAVE AUTOPSY DONE TO LOOK AT ITS LEVEL. SO, THERE, SO THERE ARE SOME CLINICAL DIFFERENCES BUT YOU KNOW IT'S NOT ABSOLUTE, IT DOES SEEM TO BE SOMETHING DIFFERENT. [ APPLAUSE ]