WELCOME TO TODAY'S WEDNESDAY AFTERNOON

LECTURE AT NATIONAL INSTITUTES

OF HEALTH.

THANK YOU FOR COMING.

MY NAME IS JUSTIN TERASKA.

I'M AN INVESTIGATOR IN THE

LABORATORY OF MOLECULAR

BIOPHYSICS IN THE NATIONAL HEART

LUNG AND BLOOD INSTITUTE.

IT'S MY PLEASURE TO INTRODUCE

DR. JAMES ROTHMAN AS TODAY'S

WALS SPEAKER.

DR. ROTHMAN IS WALS PROFESSOR OF

BIOMEDICAL SCIENCES AND CHAIR OF

DEPARTMENT OF BIOMEDICAL AT

YALE.

HE'S ONE OF THE MOST INNOVATIVE

AND INFLUENTIAL CELL BIOLOGISTS

AND BUY CHEMISTS WORKING OFFER

THE LAST FEW DECADES.

DURING TENURES AT STANFORD,

PRINCETON, INVENTORY DEBT R,

COLUMBIA AN YALE, -- CLONE

KETTERING AND YALE, THIS IS

INCLUDED SEMINOLE DISCOVERIES

RELATED TO HOW PROTEINS INSERT

INTO MEMBRANES.

HOW VESICALES TRAFFIC THROUGH

THE CELL, AND HOW VESICALES FUSE

WITH THE MEMBRANE, A PROCESS

CALLED EXOCYTOSIS.

ASIDE FROM THE BIOLOGICAL

DISCOVERIES HIS LAB HAS INVENTED

IMPORTANT METHODS INCLUDING IN

VITRO RECONSTITUTION OF VESICAL

TRAFFICKING PATHWAYS WHICH HAS

REALLY ALLOWED THE COMPLEX STEPS

OF VESICAL TRAFFICKING TO BE

TEASED APART AT THEIR MOST

FUNDAMENTAL LEVEL H.

ADDITIONALLY DEVELOPED NOVEL PH

FLUORESCENCE PROTEINS THAT ALLOW

INDIVIDUAL SYNAPSES AND SINGLE

VESICALES TO BE WATCHED IN

LIVING CELLS IN REAL TIME.

IN PARTICULAR, DR. ROTHMAN HAS

HELPED TO ESTABLISH THE SNARE

HYPOTHESIS OF MEMBRANE FUSION

WHICH PROPOSES THAT THE CORRECT

PAIRING OF ALPHA HELICAL

PROTEINS ON TWO OPPOSED

MEMBRANES DIRECTS AND CATALYZES

THEIR FUSION.

ALONG WITH THESE IMPORTANT

DISCOVERIES DR. ROTHMAN MENTORED

AND TRAINED MANY PROMINENT

SUCCESSFUL BIOCHEMISTS AND CELL

BIOLOGISTS.

HE'S A MEMBER OF THE NAB

NATIONAL ACADEMY, INSTITUTE OF

MEDICINE AND RECIPIENT OF

NUMEROUS AWARDS INCLUDING LASTER

AWARD FOR BASIC SCIENCE AND

CAVALI PRIZE FOR NEUROSCIENCE.

HE WILL DISCUSS HIS RECENT WORK

ON SNARES AND THE ACCESSORY

PROTEINS THAT DIRECT THEIR

FUSION.

IN A TALK TITLED MOLECULAR

MECHANISMS OF SYNCHRONOUS

NEUROTRANSMITTER RELEASE. AFTER

THE SEMINAR THERE WILL BE A

RECEPTION IN THE LIBRARY SO

PLEASE COME AND THEY'LL GIVE YOU

AN OPPORTUNITY TO SPEAK MORE

INFORMALLY WITH DR. ROTHMAN.

JOIN ME GIVING A WARM WELCOME TO

JIM ROTHMAN.

[APPLAUSE]

>> THANK YOU.

THANKS FOR ARRANGING THE DAY,

BEING A GREAT HOST AND ALSO

SHARING WITH ME YOUR RECENT AND

EXCITING WORK especially on

novel methods used Ford

measuring confirmational changes

in proximity with fret.

IT'SER EXCITING, I HOPE WE CAN

COLLABORATE AS A RESULT OF THAT.

PLEASURE TO BE HERE WITH YOU

TODAY.

I'M GOING TO AS THE TITLE

SUGGESTS, TALK ABOUT THE PROCESS

OF SYNCHRONOUS NEUROTRANSMITTER

RELEASE.

THERE'S BEEN A LOT OF PROGRESS

OVER THE LAST FIVE YEARS I WOULD

SAY ESPECIALLY IN BEGINNING TO

UNDERSTAND THE MECHANISM BY

WHICH THIS VERY IMPORTANT

PHYSIOLOGICAL PROCESS OCCURS IN

STRUCTURAL AND BIOCHEMICAL

TERMS.

SO WHAT I WOULD LIKE TO DO TODAY

IS OFFER SOME HISTORICAL

BACKGROUNDS TO THE THE PROBLEMS

AND THEN AFTER THAT SHARE WITH

YOU OUR CURRENT VIEW IN THE FORM

OF A MODEL OF HOW A SYNCHRONOUS

TRANSMISSION NEUROTRANSMITTER

RELEASE MAY WORK.

A STRUCTURAL BUOY CHEMICAL MODEL

AND AFTER THAT SHOW YOU SOME OF

THE EVIDENCE ACCUMULATED FOR THE

MODEL OVER THE LAST TWO OR THREE

YEARS ESPECIALLY.

WHAT DO I MEAN BY SYNCHRONOUS

NEUROTRANSMITTER RELEASE?

IT'S ACTUALLY THE PHYSIOLOGISTS

HAVE VARIOUS COMPLEX SCHEMES TO

MEASURE IT AND WAYS OF DEFINING

IT.

TO ME IT'S VERY SIMPLE BUT

FUNDAMENTAL CONCEPT IN

NEUROSCIENCE WHICH IS WHEN THE

ACTION POTENTIAL COMES DOWN THE

END OF THE NERVE AND YOU RELEASE

A NEUROTRANSMITTER ACROSS A

SYNAPSE TO THE NEXT NERVE OR

PERHAPS MUSCLE CELL, THE

NEUROTRANSMITTER NEEDS TO BE

RELEASED AT THE RIGHT TIME.

WE DIDN'T WANT THE

NEUROTRANSMITTER TO BE RELEASED

ASYNCHRONOUSLY ON ITS OWN ACCORD

BECAUSE IN THAT CASE IT'S A

FALSE ALARM.

YOU ALSO DON'T WANT THE

NEUROTRANSMITTER TO GO NOT BEING

RELEASED BECAUSE YOU MISSED AN

IMPORTANT SIGNAL.

IN FACT WHAT YOU WANT, YOU WANT

THE NEUROTRANSMITTER TO BE

RELEASED PRECISELY SYNCHRONOUSLY

WITH THE ARRIVAL OF ACTION

POTENTIAL AT SYNAPTIC TERMINAL.

THE WAY THAT'S ACHIEVEED IS

THROUGH MEMBRANE GATED CALCIUM

CHANNELS, PROBABLY EVERYONE

KNOWS THAT ARE LOCALIZED IN THE

SYNAPTIC PRE-SYNAPTIC REGION

THAT OPENED UP THE GATE FOR

CALCIUM ENTRY AND IT ACTS AS A

SECOND MESSENGER TO TRIGGER

RELEASE OF NEUROTRANSMITTER

STORED IN VESICALES.

THE PROBLEM THAT WE HAVE TAKEN

ON THAT I WOULD LIKE TO ADDRESS

HERE IS NUMBER ONE HOW ARE THESE

VESICLES RELEASED?

HOW DO THEY FUSE WITH THE

SURROUNDING MEMBRANE HAVING

STORED THE NEUROTRANSMITTER

WITHIN THEMSELVES.

HOW DO THEY DO IT SO RAPIDLY?

SO MUCH MORE RAPIDLY, ORDERS OF

THE MAGNITUDE MORE RAPIDLY.

THAN OTHER MEMBRANE FUSION

PROCESSES THAT TAKE PLACE IN THE

CELL.

SO THOSE ARE THE ASPECTS BUILT

INTO SIN CROW IN THISTY.

IT'S OBVIOUSLY IMPORTANT AT A

GROSS LEVEL.

IN YOUR BRAIN IF YOUR

NEUROTRANSMITTERS WERE RELEASED

HEALTHER SETTLER, THERE WOULD BE

NO POSSIBILITY OF C9

INFORMATION PROCESSING OR

ANYTHING OF ANY REMOTE INTEREST.

YOU WOULD HAVE A -- THINK ABOUT

WHAT EXPERIENCE WOULD BE IF YOUR

VESICALES ALL FUSED AS THEY

SHOULD BECAUSE THE FUSION

PROTEINS ARE CONSTITUTIVELY

ACTIVE.

THEY SHOULD FUSE AND RELEASE

NEUROTRANSMITTER ALL AT ONCE SO

THEN YOU HAVE EVERY

NEUROTRANSMISSION TAKING PLACE

AT ONCE WITHIN A SHORT PERIOD OF

TIME, NO THOUGHTS WHATSOEVER OR

PERHAPS EVERY THOUGHT YOU WOULD

HAVE AND NOT BE ABLE TO

COMMUNICATE TO EVERYONE ELSE, IT

MIGHT BE AN INTENSELY

TRANSFORMATIONAL EXPERIENCE BUT

IT WOULD BE ONE THAT LASTS 10

MILLISECONDS.

SO THAT CLEARLY DOESN'T HAPPEN.

IN A MUCH MORE SUBTLE LEVEL THE

SPEED OF SYNAPTIC TRANSMISSION

IS VERY IMPORTANT FOR THE

COMPLEX CIRCUITS THAT WE HAVE.

IT TYPICALLY TAKES A FEW

MILLISECONDS FOR A SIGNAL TO BE

TRANSPORTED ACROSS A SYNAPSE OF

WHICH THE RELEASE PROCESS,

INITIATION OF IT TAKES TYPICALLY

LESS THAN A MILLISECOND IN

CENTRAL SYNAPSES.

THAT'S ACTUALLY VERY IMPORTANT

BECAUSE FROM THE TIME THAT A

PRIMARY PIECE OF POTENTIAL

COGNITIVE INFORMATION LIKE

VISUAL FIELD AND AUDITORY

PATTERN WHAT HAVE YOU, IS

SENTENCED BY OUR INPUT OUTPUT

DEVICES, IT HAS MAYBE 20 OR 30

MILLISECONDS FOR ALL THOSE

PATTERNS COALESCE AT YOUR

HIGHEST CENTERS HAVING BEEN

TRANSMITTED THROUGH PERHAPS TEN

OR 15 DIFFERENT SYNAPSES.

I THINK WE'RE ALL FAMILIAR WITH

SOME ANYWAY ARE OLD ENOUGH TO

REMEMBER WHAT MOVIES USED TO BE

LIKE WHERE THE FILM GOES BY AND

YOU HAVE 35, THE MAGIC NUMBER

FRAMES PER SECOND.

WHY 35?

BECAUSE IF IT'S FASTER THAN 35

IT LOOKS LIKE A CONTINUUM TO

YOU.

IF IT'S SLOWER THAN 35, YOU SEE

SEPARATE PICTURES.

HOLLYWOOD FOLKS WEREN'T SPEND

THRIFTS THEY WERE LOOKING FOR

PROFITS, SO THEY FIGURED OUT

THAT THE GRANULARITY OF HUMAN

EXPERIENCE IS ABOUT 25 OR 30

MILLISECONDS.

SO YOU CAN GET BY WITH THE LEAST

NUMBER OF PHOTOGRAPHIC FILM AT

THAT SPEED.

SO THAT 25, 30 MILISEDGES IS

WHAT YOU HAVE THAT REPRESENTS

SIN CROW IN THISTY AND THE JOB

IS TO GET THROUGH 10, 15

SYNAPSES AND PLOW THROUGH THEM

FROM HERE AND HERE UP TO HERE.

FAST ENOUGH.

HOW DOES THAT WORK IN MOLECULAR

TERMS?

THAT'S THE PROBLEM I WOULD LIKE

TO THE ADDRESS TODAY.

THE SOLUTION TO THIS PROBLEM HAS

COME I WOULD SAY OVER SEVERAL

DECADES, ACTUALLY MORE LIKE HALF

A CENTURY, MANY WHICH CELL

BIOLOGY AND NEUROSCIENCE OR

NEUROPHYSIOLOGY AS IT WAS THEN

CALLED DOVE TAILED TOGETHER,

GONE APART COME BACK TOGETHER

AND THERE'S MANY CRITICAL COMING

TOGETHERS OF THESE TWO FIELDS.

THE BEGINNING OF THIS FIELD IN

FACT REFLECTED THAT.

GOING BACK TO THE CLASSIC WORK

OF FAT AND CAT WHO FOUND HOST

SYNAPTIC POTENTIALS, THE

NEUROMUSCULAR JUNCTION IN

CLASSICAL EXPERIMENTS FROM

1950s, IF YOU HAVE AN

ELECTRICAL IMPULSE STIMULATING

THE NERVE INNER INVESTIGATING

THE MUSCLE, WHEN YOU GET

SYNAPTIC TRANSMISSION THE

MEMBRANE POTENTIAL OF THE MUSCLE

CHANGES IN RESPONSE.

IN YOU HAVE NO INPUT NO ACTION

POTENTIAL, ON THE MUCH SMALLER

SCALE THERE'S MINIATURE

POTENTIALS OR MINIS THAT APPEAR

AT FAIRLY LOW FREQUENCY BUT YOU

CAN SEE THAT THEY'RE ABSOLUTELY

MEASURABLE AND SEEM TO BE OF

UNIFORM SIZE MEASURED AS A POST

SYNAPTIC POTENTIAL.

IN THAT CAPS THE IDEA

TRANSFORMATIONAL IDEA THAT

SYNAPTIC TRANSMISSION THE

RELEASE OF NEUROTRANSMITTERS

OKAY CANS IN A JUAN UNTIL

FASHION WHICH THERE ARE

INDIVIDUAL PACKETS OF

NEUROTRANSMITTER SOMEHOW

PREARRANGED STORED AT A NERVE

ENDING SHOWN HERE.

ABILITY THE SAME TIME IN 1950s

MY PREDECESSOR AND FOUNDER OF

CELL BIOLOGY AT YALE, THE

FOUNDER TO LARGE DEGREE FIELD OF

CELL BIOLOGY OBSERVED THESE

MEMBRANE ENCLOSED VESICALES AT

NERVE ENDINGS.

HE CALLED THESE SYNAPTIC