A Conversation with Paul Greengard

I'm Eric nestler I'm at uh Mount siai School of Medicine in New York City and it's my great honor to be here today to interview Dr Paul greenguard we're sitting in Paul's office at the Rockefeller University I had the privilege of being a graduate student in Paul's lab in the late 1970s and early 1980s so it's particular fun to be here today uh Paul thank you for joining us um thank you Eric I thought we'd start By talking about the contributions that your laboratory has made to the field of neuropharmacology and start with the early

days when your lab started looking at the role of P CYCC nucleotides in the brain what prompted you to take that Focus uh what prompted that was the discovery by earol suland and his colleagues that the hormones Norine and glucagon raised the level of Psych in liver and muscle cells and the thought occurred to me that Maybe nerve cells communicate with each other in a manner similar to that by which the hormones communicated from a sending cell to a receiving cell despite the difference that the hormones are working over matter of in some cases feet

M and these things are through millions of a centimeter MH uh and I remember having that thought many years before I did anything about it I was doing my uh postdoctoral studies and then I was in industry for Uh nine years pharmaceutical industry so when I got the first thing I did when I went to Yo was to see whether neur trans might raise psychic and we found that there several ad doine sens Dental cycl octopamine sensitive D cycl serotonin sensitive dental cycl and it was clear then that the principle that the endocrine system had

evolved was also true for synaptic communication across a tiny synaps MH and then you also took analogies from the earlier Studies in endocrinology to pursue how it was the cyclic nucleotides then produced their functional effects in nerve cells uh with the discovery of protein kinases in the brain right that that became clear many years after the subin discoveries when as a paper by WS Perkinson Krebs showed that the psych Works through an enzyme which at the time was called phospher kyes kyes and the name was unchanged to psych Ependent kyes and I wondered whether the

same thing might happen in the brain that there was a kinas involved in mediating the acents of neurotransmitters and so we looked for such a such an enzyme in the brain and we found there was in present in much higher concentrations than anywhere else and in addition it was associated with the plasma membrane so everything was in the right positions to make that a viable hypothesis that protein kindest Medi the actions of neurotransmitters and uh the argument about whether psychia was important was finally resolved we published two papers back to back PES mhm one with

Felix traser calch and the other one with Eric kandel at Columbia in which we showed that injecting the catalytic subunit of Psych and P dependent peries into neur cells mimic the effect of the neurotransmitters so that was pretty Definitive evidence I was in your lab at the time that those studies were published and I remember uh being struck by the reaction of the field to your lab's proposal that cycle GM and cycle GM dependent protein kinas are important in regulating neural phenomena there was a lot of skepticism would you like to uh talk about that

the peak of the skepticism came when I gave a lecture in front of a very large audience and after I finished my talk somebody got up and Said this is absolutely heretical thinking and should not be allowed I thought that was a little bit excessive but that that was the bottom but then gradually people be began to accept it I think was there a turning point you think that um or was it just more of a gradual process I think it was a gradual thing there was good news and bad news about and people not

accepting this the the good news was that I didn't have any competition for 15 years from the late 80 late 60s to the early 80s uh the bad news is was I was sort of unpopular conceptually in the field mhm but the good news outweigh the bad news so we're able to develop this story incrementally but over over that 15 years the evidence became overwhelming and it was hard any longer to deny it ironically the first place we showed the most compelling evidence that neurotransmitters could work through these Pathways we were we're Looking at nucle

aryes bird bird arthro sites and were're able to show that when they neurot transmit I believe was neurop nephrine I'm not sure activated its Target soell there was an increase in psym in those cells and the psych could produce the effect on the Ion channel could mimic the neurotransmitter producing that that was pretty compelling then we went on to show the same thing in nerve cells mhm it was a gradual buildup and then we were looking For substrates for this pathway and found a few of those and found other Coes so increasing the the evidence

became stronger and stronger M I'd like to ask you about those other kinases cuz I remember that being part of the process by which your lab established an important role for protein phosphorilation after the finding of cycle GMP dependent protein kise in the brain at high levels I believe your lab went on to discover cyc GMP dependent Protein kise that's correct the way we found that I wanted to purify the psych page so put this extract over a column mhm and we got our two peaks and at that point people have gotten very interested in

other psychic nucleotides look like Psy GMP psychic all [Music] the and so we had this second Peak and so we compared the two peaks and found that psych activated the kyes activity of the First Peak and psychic GMP activated the activity of the second Peak and then we by dissecting that system realized it was a totally different protein canies regulated by psychic GMP and not psychic and that too became a controversial issue it's interesting in retrospect some of the people down at Vander who have been fellow students if you will of earol suland were really

didn't like this they said this is some M some denatured form of Psych and pedop Kindness which now s psychic GMP dependent kindness as the activator not psychic as as the activator so mhm it was interesting absolutely and then shortly thereafter your lab discovered what has come to be called uh calcium codul independent protein kise 2 among other calcium kinases N1 yeah yeah it was very similar three I believe perhaps right yeah no we didn't have three okay that came later but it was the same thing we knew there was Cal dependent Protein phosphorilation so

we want to see how that was happening and what we did we ran an extract over a column and found there were two peaks of calcium dependent kindness activity present and the one we call calcium dependent protein what we Chang the name later to calcium C dependent protein one and Cal C independent per k 2 today we know that cam k 2 as is abbreviated is much more important it's an important component of the postep density it plays A vital role in syneptic transmission mhm I think it's hard for students who grow up in biomedical

science today who by second nature know that protein phosphorilation mediates so many different neural phenomena to think back not that long ago really 30 or so years ago remember you published a paper in a review in science in 78 where I believe one of the first times you proposed A diversity of kinases with mediating A diversity of types of signals on many Types of neural phenomena right um and now the human genome uh sequencing has indicated there's about 500 or so protein kinases so I don't know if you want to comment on that perspective but

there's now over a thousand tyrosine protein kind which is one subclass right um I think just a lot of evidence accumulated not only in our lab in many other Laboratories and and now with the modern molecular bio we could knock out genes or mutate them and so on and you Know the evidence has became incontrovertible and now as you say it is second nature to [Music] people I mean it really has transformed I think the way young people approach a biological question because protein phosphorilation is almost a reflex in terms of a mechanism that's pursued

yeah now that's was come a long way from the see at the time we started this work it was accepted that Chemicals the the way neuros cells communicated with with with each other was through neurotransmitter release diffusion to the post synaptic membrane to activate receptors which at the time were hypothetic of when we started this and that was a battle that was went on for 40 years and it was won pretty resoundingly by the chemical School a very distinguished scientist such as Eckles who took a long time to Accept it m believed in the electrical

transmission which is perfectly logical the idea was that when the nerve impulse came down to the ending that was electrical field generated by the nerve impulse which changed the membrane potential across the post synaptic membrane and initiated or inhibited a post aptive response but as I said the debate was one rather resoundingly by the chemical school was about this time we started Our work and and at that time people believe okay it's a chemical but it's an iCal response to an opening of a voltage Gate iron Channel which initiated the signal going down the post

synaptic cell and there was reason to believe this the things that did that were for example AET coling acon receptors open up iron channels those studies were perfectly compatible what was known at the time with the idea that All that was involved was an El change in ion conductivity across the membrane and therefore an inhibitory or an excitatory postoptic potential but again as we discussed earlier I thought it might be more complex and it might be somewhat like the hormones and that turned out to be the case you can say both ideas were correct you

can think of that electrical signal as being how the fast excitatory neurotransmitter glutamate and the fast Inhibitory neurotransmitter Gaba produce their effects MH but then there are over a 100 probably many many more uh neurotransmitter Pathways which work through these very complicated signal transduction Cascades involving changing level of second messenger activation of a protein Kye or protein phosphotase changing the phosphorilation of a key substrate protein at the production of a physiological response Mhm you will lose to earlier Paul the that the next step after identifying protein kinases was to find the substrate proteins through which

the protein kinases work to produce their functional effects um uh let's talk a bit about the first substrates that your lab uh discovered in the brain uh which uh Was A protein that I was involved in we called it protein one at the time maybe you can tell us about protein one sure so um After he established that the neurotransmitters work through changing level of the second messenger and the second messengers work through activating protein kinases or protein phosphatases the next question came what are the substrates for the cois and phosphatases and those days they

had just developed these very primitive techniques for doing running gels and doing Western blotting and or and doing radio r determination radioactivity on The on the gel and what we did was we looked for uh we use radioactive ATP and look for substrates that might be radioactive and then we found this protein which we called synapson one with two of them synapson one and synaps excuse me protein one and protein 2 and it turned out as we learned more about protein one and and purified it that protein one was associated with the nerve Terminal and

as we subsequently learned it It Coats all synaptic vesicles and regulates neurotransmitter relase in a very profound and dramatic way so that was synapse in one and synaps and do and the reason in retrospect that we found them first where they're basically present on all on all syneptic vesicles and all nerve terminals in the brain and so they're enormously abundant and so with the Primitive techniques that were available at that time they were the first ones we saw because they were the most abundant right and in fact the discovery of synapson I think and demonstrating

its concentration in nerve Terminals and its role in neurotransmitter release was very important in stimulating the field to elaborate a biochemical process of neurotransmitter release which had before that Discovery remained completely opaque Right so yeah that's true once we found out this that this synap protein one was synaps and one which got phosphorated then we compared what synapse in one in the phosphorated form did compared to the def phosphorated form and what we found was that the defil form of synaps one inhibited the vesicles from fusing to the plasma membrane and releasing the neurotransmitter the

phosphorated form did not do that see by Fling you remove this inhibitory uh effect and allowed an increase of neurotransmitter release mhm uh and it turned out that there these two U classes of synapsin which even to this date is not quite understood why they because it's synapsis 1 a synaps 1 B 2A and 2B it's not quite clear why one needs so many of these synapses to do what the nerous cell does was cuz by and large the Ratios are pretty much the same amongst all different nerve terminals mhm Evar Wallace and Angus n

to postop laboratory took on the project the following project we knew that synapson one was present in every Nur cell in the brain we asked whether there might be certain fosil proteins phosphorated proteins which present in certain region of the brain and not in others because the brain brain is so heterogene so heterogeneous mhm I have To say that also in retrospect it's say discovery of these fosil proteins and these siging Pathways that provide un provide overwhelming proof for the heterogenity of different nerous cells like when I started out it was like a ner cells

a ner cells a ner cell despite the fact that ramoni kahal had shown these hundreds of different types and atomically what our work did was to show there 100 different types biochemically MH so it did help in that In that regard another substrate that your lab discovered a number of years ago that remained a major Focus for a while was Dar 32 uh would you describe for us what led your lab to discover that protein so we were looking for region specific foso protein and we found that we took the the neum we found a

whole bunch of these different phosphor proteum bands which You didn't see for example in the cortex the the explanation for that retrospectively was that the stum is very homogeneous compared to the cordex cortex has a lot of different cell types the strum has basically these two major classes of what are called medium spining projection neurons in the cortex there are only cell types and therefore all these different foso proteins which didn't stand out in the stum there's two cell Types basically that stand out and they these cell these two cell types which are very similar

in many respects have a large amount of this one fusso protein type and we're able to see it because of the homogeneity wasn't diluted by lots of other and the DAR 32 turned out to be very important in dopamine signaling mhm hence its name yeah that Dar 32 is an acronym for dopamine psych and P dependent py dop Pych regulated foso protein of 32,000 and more recently your lab had shown that uh the protein's role is far broader than simply mediating the effects of dopamine yes the reason we focused on the dope initially is when

we found this mod molecule selectively in the strium we go maybe was found by looking for psych pting count substrates because dopamine was such a prominent neurotransmitter There we then showed that dopamine regulated and given the clinical importance of dopamine in many psychiatric diseases Parkinson schizophrenia ADHD uh we studied the dopamine system a lot the other reason for focusing on the dop system for a decade or more actually two decades was that the concentration of Dar 32 in these principal cell types in the stum Was enormously higher than in the rest of the brain so

if you did imuno staining or imuno peroxide staining the stum was black and the rest of the brain was hardly labeled it turned out that this tiny amount of D 32 in the cortex for [Music] example was as vital for those cells in the cortex as it is for the stratum and the reason why it's even though it's present in tiny tiny concentrations the Reason it's effective in the rest of the brain is because Dar 32 uh is an incredibly powerful U molecule in its phosphorated form it inhibits prum phosphotase one the major sering 3

phosphotase in the brain mhm at 10us 10th molar which means a trace amount of Dar 32 can fully inhibit this vital enzyme the proteum phosphatase and thereby control the state of phosphorilation of hundreds of other proteins right right Mhm one of the interesting historical findings I think was the observation that several of these substrates like synaps and like darp 32 are phosphorilated by several kinases at either the same or different sites and at the time I think it revealed a completely new level of molecular convergence of Regulation and I want to talk about the implications

of that perhaps yeah okay so it's true we we found each one had his own story but we Found several phosphor sites for different kindies there were four major ones and now we know three minor ones which we've barely begun to mine but the four major ones this is on Dar 32 on Dar 32 and we found that these four major phosphorilation sites on Dar 32 they were phosphorated by different protein chineses in response to different neurotransmitters so it became clear was this is a way of these different neurotransmitter Pathways interacting in The DAR 32

containing cells and one of the ways they interact is very simple when one of these the the the psych and P site the Dolan psych and P site was a residue called 3 34 mhm and when that is phosphorated you got this very potent inhibitor protein phosphotase one mhm the other three sites were phosphorated the consequence of phosphor on those other sites was to affect the uh efficiency of either phosphor or def Phosphation of this 3 and 34 MH so that the other neurotransmitters are now modulating the doine signaling pathway through this complicated int molecular

mhm uh interaction in the dark 32 molecule yeah leading to very complicated modes of cellular regulation right it's just a fascinating Story how these uh biochemical networks of information control have evolved yes so it it turns out that this is the major a major mechanism Not the major mechanism by which these neuro different neurotransmitter Pathways interact with each other so got this convergence of all the signaling onto a single molecule the D 32 Through The neurotrans receptors second messengers Kinesis onto dark 32 and the dark 32 in turn by regulating the activity of protein phosphation

One controls the state of phosphorilation virtually every Downstream protein all the effect of proteins remarkably this Hardly a protein in the brain that exists that is not regulated by protein phosphotase one M so so you have this convergence onto the dark 32 and then a Divergence going out from that interesting we found another molecule recently which uh plays the same role for calcium that the dart 32 place for psych MP and that's a molecule we call regulator of calmodulin signaling and that also integrates a whole bunch of stuff and Interacts with a bunch of stuff

that goes through the psych and P people might know that as RCs protein regul C RCS was also we found it in the same original studies that we talked about earlier when we looking for substrates for psych dependent crus that might be region specific we found the several bands on a gel I mentioned that earlier had several molecular about six of them very interesting we so consumed with an AR 32 for 15 years that we didn't go to These then we turned to those other and we found another one of these it was ARP 20

one or Dar 21 MH and that was regulated by the calcium pathway and interesting this RCS or uh AR 21 controls the state of Def phosphorilation One of the reges on Dar 32 so it's a regulator of this master regulator molecule it's a way was one of the mechanism by which the calcium sing pathway controls the psych signic pathid Paul let's turn to the focus of Your lab in more recent years uh when you have when a major uh interest of the group has been Alzheimer's disease okay maybe you can describe a little bit about

what Drew you to focus on Alzheimer's disease and in particular talk about your recent discovery of a major protein that controls amalo uh deposition in the brain all right so I mentioned a few minutes ago because of this vastly increased amount of information we had about signal Transductional pathways uh I thought it might be possible to understand the molecular basis for various neurological and psychiatric disorders such as depression um schizophrenia Parkinson's and Alzheimer's MH and to try to gain insight into the mechanism by which the drugs that used to treat those disorders achieve their effects

with regard to your specific question about Alzheimer's it was known that beta amalo was made in in the brain by converting a molecule called the amalo precursor protein into an intermediate amid precursor protein C terminal fragment and That Was Then converted to Beta amid these are two steps requiring two enzymes known as beta secret a and Gamma secret a MH so we were able taking cell line into a cells to permeabilize them and show that we could still make beta amid in these Broken cell preparations but that was exciting because we can now start controlling

characterizing the enzymes so we now had these broken cell preparations we found that they could make the beta amid in this two-step Cascade we were then able to dialy all the interior of the cell out and show you loss of the ability to bake beta amid MH if you added back ATP ad Denine triphosphate you're able to accelerate The conversion of the AP PP the amid precursor protein to Beta amid we then showed that was due to the Second Step the gamma secr days and then we kind of left that on the shelf for several

years and then a post doal fellow in lab bill ner picked up the gauntlet on this and wanted to see what the atps working on and so the way this project was approached was to uh take drugs which are known to work by Competing with ATP at binding sites this is mainly these tarine kindness Inhibitors which are used very extensively now in developing anti-cancer drugs MH and one of them glec or aatb which is made not by novaris and is now used to treat people uh with the chronic myogenous leukemia they this GAC was able

to prevent the ability of ATP adenine triphosphate to catalyze the conversion Of the a the amalo precursor protein to bet Amoy we published that I was a bit concerned about it but these younger people have to get their papers out I was concerned about it because I thought once we showed this that this leac could block the formation of beta Amo that the whole pharmaceutical industry would all come in and they didn't part of the reason for that was one pharmaceutical company published a couple of papers Saying this is an artifact which we knew what

it wasn't but it bought us time it kept other people from mudding the subject MH and so we went on with this and we were able to show that the GIC bound to a protein which we today call gamma secretase activating protein or abbreviated Gap mhm an acronym what happens is that is a is turned out that the this Gap or gamma C activating protein binds to the C terminal fragment of the AP molecule it Then forms a turny complex of the gamma secra so you have the enzyme the substrate in this modulator the gap

which activates the cleavage to form beta amid this one of the problems that developing Inhibitors of gamma secat to block beta Amo information is they cleave some other pathways most notably Notch and so most of these gamma secret Inhibitors have not only by inhibiting gamma secretes not only prevented the Formation of the bad substance betoid but they've also prevented the formation of vital substances fortunately with a gap it only affects activates only the bet amid pathway not the notch pathway and that was great because it means that Inhibitors of Gap M can block beta amid

formation without blocking Notch formation and if you put that in a broader perspective what what is telling us is that the Gap is acting like many There let me back up there are many regulatory proteins known now which regate proteases proteases are very promiscuous but they require a regulatory protein to say attack this substrate and not this one and that's what the case is with gap is exactly a classical regulator of proteas mhm the GP regulates the ability of the gamma secretas to cleave this a CF to form beta amid it does not affect the

notch pathway so now there's a whole new area It's an exciting new area for drug Discovery for where you can look at Inhibitors of Gap and we have made compounds which are very potent selective Inhibitors of beta Amid and not of not affecting the notch pathway and we' made knock out mice of Gap and they no longer form plaques they no longer form these beta the amount of beta amul is greatly reduced there are no plaqu formed in the brain but there's no Toxicity either as you can look at so kind of intestinal pathology they

looked at when you inhibit the to look for Notch related not of the notch nicd it's called so this is a nice selective thing and we're now making compounds which are much more potent uh Inhibitors of the gamma pathway selectively for the beta amid so I think this is going to be a very potent way to get into exciting new drug development We have compounds which are more potent on the in terms of interacting with G oh I ne I neglected to to mention that the in this trinary complex of the Gap the gamma CRA

and the AP what happens is that the ATP is required it causes a confirmational change in the gap which enables it to activate the cleavage and that the GLE competes with the ATV binding site on the Gap and we' just now identified the residue of the ATP BS to so we're very excited which Will help drug Discovery even even further more one last Discovery I'd like you to talk about before we uh discuss other subjects is your lab's focus on a protein called P11 and uh which you've implicated in depression leading to potential new therapies

for that syndrome as well could you tell us how your lab came across P11 yes uh it's pretty well established that Virtually all well certainly the ssris the Ser selector seratone reuptake inors but even some of the other anti-depressants like the monomin oxidase Inhibitors and the tricyclic anti-depressants appear to have their anti-depressant effect through increasing signaling of Serotonin synapsis and they do this by increasing amount of serotonin in the synapsis not affecting the post synaptic serotonin Receptor we wondered if and in this case this is the work of pis fennings and Mark flagel MH uh

we wondered whether there might be endogenous proteins in the plasma membrane of the serono septo cells which might modulate the serotonin receptor so yeast using yeast two hybrid technology which Mark FL had introduced into our lab he had done his Graduate Studies in at the pastor Institute in a laboratory that was one of the major developments Of east2 hyper technology so when he came here he started working with peir on this project can we using the serotone receptor find a binding protein which might regulate its activity M and he found this protein P11 in contrast

to the gap which is totally unknown it was not even in the human genome at the time we found it the P11 was a well-known protein but there was no evidence that it was involved in brain function mhm so here we found this P11 to be a binding protein in the plasma membrane for the serot for certain serotonin receptors not all and so then the question was is this a bi possible biological relevance and we found that it cooll localized with the the serotonin receptors or certain of them and then we found most excitingly that

the P11 was a chameron which traffic it caused accumulation of the seron receptor in the plasma Membrane we still don't know whether it's due to an increased Rec mment to the membrane or decreased endocytosis M but that's one of the things we're working on mhm now this was terribly exciting because all of the previously known anti-depressants work by increasing serotonergic transmission by increasing the level of the neurotransmitter serotonin with no effect on the receptor here we had a protein which did the opposite it Increased the amount of the serotonin receptor in the presence of fixed

amount of the neurotransmitter serotonin mhm so this seemed very exciting and we wondered whether in fact there might be a relationship we showed in a number of ways for example changing the depressive state or an anti-depressant state of animals caused a change in the P1 levels for example uh uh in animal models of depression MH the Level of P1 was way down postmortem human brain tissue depressed people major depressive da had a much lower level of P11 than normal controls conversely we can manipulate the state of P1 and get these really dramatic behavioral effects for

example if we knocked out the P11 the animals had a major depression as judged by all of these Behavioral Studies a lot of people who are not familiar with the Field say how can you judge from a mouse's Behavior or rat's Behavior uh whether this is a good predictor of depression in a human the fact is there's a good correlation as you know well Eric from your own work there's a good correlation between the ability of different drugs to be clinically effective and their ability to do these uh to have these effects on behavior in

rodents MH so we knocked out P1 constitutively and got this depressive Behavior mhm if we overexpressed P1 the animals behaved as though they'd been given an anti-depressant mhm we then H in a series of studies with Michael kaplet who had been an mdphd student Here and Now brain surgeon at cross the street of Cornell mhm we did studies to localize the region of the brain at which this might Occur we found if you did not the consti knock out but only knocked out the P11 and the nucleus cumin a known system for reward and and

uh mood mood uh regulation if you knock that only in that region you got a depressed animal we then went on to show that within that region this is the work of Jennifer Warner Schmidt if you knocked out the P1 only in one cell type in this relatively small region of the brain the nucleus of Comin you mimicked the Constitutive knockout we were able to identify a single nerve cell type where this happened and in fact it's a minority of cells in this entire region right it represents mediating this effect 1% of the cells in

a small region of the brain you remove P1 you have a severe depressive phenotype we're very excited about this we're able to take instinctively knocked out mice so there's no P11 anywhere in the brain or body and put The P11 back into this one cell type mhm these uh inter neurons that represent 1% of the cells in the nucle Gs and totally restore normal activity so this has been a rather rewarding series of experiments we now know of P11 enriched cells in several parts of the brain interestingly uh there in in studies in collaboration with

Eric Schmid a post octal fellow in N Height's laboratory and Jennifer Warman in our laboratory uh we' just Found in a paper in in that just came out in Cell last week that there's a cell type in the cortex in layer 5A in the CeX which is highly enrich in P11 if you knock out the P11 there you what you do you you you get the opposite of what you got if you knock out the nucleus of com you get and if you knock it out in the cortex in these layer 5A cells the animals

show no depressive phenotype but they no longer respond to an anti-depressant substance With a behavioral change this is in contrast to nucleus of cumin where if you knocked it out in this one cell type they had a depressive Behavior but they retain the ability to respond to the anti-depressant so one of the things we're doing now is seeing if there's a possible connection between these two cell types in other words given anti-depressant it overcomes the damage you did from the knocking out in the Nucleus thatum there has to be a circuitry that involves a maybe

third or fourth or fifth class of but that's the way we're going with that MH and if you think of it it's quite analog to some other diseases Parkins disease is it represents the death of doine producing cells you treat that disease right now by compensating for the missing dopamine and treating dopamine receptor cells and this would be quite the same with these neurons in The nucas acumin are the uh reason for the depression but you compensate by uh increasing activity these P11 cells in the cortex mhm so we've had a conversation that spans 50

years of contributions from your lab and I'd like to turn our attention to address a slightly different question and that is more the social aspects of Science and then in addition to all of these Legion contributions that your lab has made Over the years another contribution you've made is to train generations of additional scientists I was if you wanted to comment about that aspect of looking back and uh thinking of the scientists who got their start with you yeah I'd be happy to that one of the things I'm happiest about in my career is that

I've Had The Good Fortune to have so many brilliant young people now many people would say the people such as yourself and you Know couple of dozen others um what I did for you I trained you to be great scientist and you did no I don't think I that's more credit than I deserve I think all these brilliant young people realized how important what we were doing was and they came to the lab because they were brilliant enough to realize how important it was I've noticed over and over again it's always the younger People get

the really important advances for example there several times in my career despite my experience along this line with some young person has come along with something a new way of looking at brain function I'll say to myself that can't be right this doesn't feel right it doesn't fit into my thinking about the brain and they were right and I think that's what happened with me a lot of older people didn't believe in it but the younger people did And so they came and studied with me and learned our trade and went off and did their

own brilliant stuff but I didn't turn them in I might have helped them be more effective s is but they had Brilliant Minds to begin with like you thank you but one of the things I I still uh after all these years of being in the lab and I think it's been over 30 years that I've worked in your lab will still go back to things that I learned from you not only about how to analyze Scientific data which is also true but in particular how to work with the younger scientists in my lab and

uh and get them to be and get them to reach their own potential you were L good at it tell me what I did cuz I don't it must be a critical ingredient that I have been trying consciously and purposely to identify and I'm not sure that I've yet identified it well I think in my case it's a problem because first of all there a correlation Between the talent of the people and how often they want to talk to me you think it'd be the opposite the the neediest students in post talks would come more

often they don't MH the ones that are most Talent would love to come and talk about their work with me MH so that's one observation which I wouldn't have predicted to be the case I think the problem is when you have a very talented person and they want to go down this path and you think It's better go down that path should you let them go down the first path rather than the second path and I tend to do that and it's usually wrong because I've had so much experience that you develop an intuitive sense

of what are the more productive a Avenues but once in a while they were right and it's you know been led to very exciting stuff so I don't think one can generalize I don't know how you solve the problem I hav't Sol no I would agree actually I think that that makes a lot of sense yeah it's also made your lab very Dynamic from the point of view thinking back to what we were talking about earlier and the resistance of the field of Neuroscience to some of your initial discoveries your lab has not resisted new

discoveries your lab has continued to embrace new discoveries new technologies to a very impressive degree and thereby remained at The Cutting Edge yeah but maybe I shouldn't Have said no that can't be right I should have said I don't think that's right but let's see whether it's relevant to what we're studying yeah let's talk also about your early years in science and and what got you uh started yourself uh one of the things that I realized when I was your GR graduate student was how unusual it was for somebody to go to take your path

to go from industry and then having a spectacular career in Academia I've Known many people who've done the opposite and I was wondering if you could take us through that early part of your career what first prompted you to go to Industry and then what led you to go back to Academia all right I went into industry because I was always excited since I was a student with the idea that one might take one's knowledge of basic science to develop drugs tell people Um and it seemed a very exciting possible way to go and then

I was asked to become the head of this division of what at the time was gagi then they merged and became cagi and then they merged it with no with Sandos and became novaris mhm and so I went roughly when did you go there was that late 50s or early 60s 1959 Mhm yeah uh I actually officially joined them in 1958 I spent five postal years of studying in England and I went I joined the gagi at 1958 and was there 67 but the first year the Laboratories weren't ready so I wor went down and

work with Sydney uden friend at the NIH mhm uh and then I went into the industry and you I was in a pretty senior position for a young person and uh as AE Of one of the Departments there were three departments there was a biochemistry department they they hardly use the term electrobiology then there's a pharmacology Department a CH synthetic chemistry department and then a medical department and then there was somebody who was like premus interp powerus amongst that he kind of let the thing but every time I had not yeah just about every time

I had a new idea about how to Develop some drug uh you had to you couldn't start a program unless the committee agreed and virtually the committee never agreed some of those I did anyhow and they look very but I got so frustrated by the time that I left in 1967 I just said I think I have a better chance of contributing to drug development Academia than I did here at gagi So In fairness the situation is very different now there Many more exciting opportunities now in those days the synthetic chemists ruled Supreme they were

the gods and all they wanted to do in those days was to make Chemical modifications to somebody else's patented drug MH so that we could get a drug that would make more money there was no biological thinking at all and I didn't realize that they would be so obtuse now the situation is very different and usually it's biologist in charge now and And in those days the when I was there the biologist were the Servants of the chemists and now it's kind of turned around and then when you made the transition back to Academia you

did so through a series of sabatical no I had I had been uh the last three years of my five years in England I was studying at the National Institute for medical research and there two guys there named Murdock Richie and Bill Douglas who were both Prominent yes pharmacologist they moved to Einstein about the same time that I moved to gagi I I really loved him and it's a peak of McCarthyism and it wasn't so pleasant and I'm not politically active but I I think and I like to see people I knew were being investigated

by the McCarthy committee it was kind of people forget how it was really unpleasant and I enjoyed England and I'm I'm kind of a Non boastful type and I and English tend to be non- boastful so I felt more comfortable there than competing with the boers here so I really loved it there and uh and uh but then the after Sputnik money started really going into in the US US science and it didn't Eng so for example I had had to develop my own florom to develop a very powerful new method for fing what we

called intermediary metabolics so I decided I had to come back to the States For professional reasons so I I worked at um at gyi for these eight years and then uh I decided to leave and I took a job at a place which wasn't built yet on Staten Island n New York State Institute for mental retardation I think they've changed the name mhm but it wasn't built yet so I had a sabatical year basically uh and I spent six months at Albert Einstein and six months at V Alber said Murdoch Richie and I my old

colleague from England and I were working was Murdoch the chair of pharmacology Al Gilman was the chairman of pharmacology and it was really probably the best department of the country that time and Mur worked out the mechanism of action of local anesthetics uh in fact I was just subject to an interview about how we discovered that we we showed that the local anesthetics in Cross the Plasma membrane of the axone in the neutralized form and the acidic into acidic M inside the axone made them protonated and so they they couldn't get out and they sat

in the sodium Channel and blocked it which this a fun thing anyhow at about the time that I was leaving U gagi to go to the Staten Island Institute uh Richie and Douglas were asked to be finalists for the chair of pharmac col at Yale which Arnold Welsh He was a cancer chemotherapy guy and they asked Richie what his conditions were he said if I will accept the chairmanship I can bring Douglas in greeng guard and dougas said I'll accept the chairmanship I can bring Richie in greeng guard so at this point they overed me

this actually Richie one the was the one called because when Douglas was asked what he would do he was asked by the then Dean what would you do about this Dead wood in the department uh he said I'll tell you what I would do with them these there like six professors of chemotherapy say I put them on a raft I get in a motorboat and I told them after to the middle of Long Island sand and i' cut the rope and come back to New York to New Haven Harbor and I always wonder why Richie

got the chairmanship over him but I know from the dean that was the reason it's funny so uh having had both of them teach me Farmacology I I could see Bill Douglas doing that can't you see that yeah that was Bill Douglas um and then yeah that's how I ended up at Yale so that's how you ended up and then you moved to I almost went a year earlier mhm so you moved to Yale in 68 yeah you almost moved a year earlier there there was a point that was going to be in the secur

department and there some people strongly for me but one powerful guy opposed to me he said no what does this have to do with Psychiatry yeah basically yeah he's a very well-known guy whose name I will not no but you proved but you proved yourself correct and them wrong him wrong him wrong let me ask you uh some even earlier questions in your training uh you were born in New York City yes uh what high school did you go to I went to two high schools I went to for excuse me first I went to

Richmond Hill High School mhm and after two years they had started Forest Hills High School and then I transferred there so you grew up in Queens Forest Hills Queens I spent first 10 years of my life going up Brooklyn then moved to Forest Hills okay and they had very few high schools then we had to it was a long long walk to go to Richmond Hill High School it's not quite like Abe Lincoln but wasn't so good there was electricity then there was electricity uh but then after the two years of Richmond Hill High School

they Built the farest Hills High School so I went to that one MH that's a school which remember Simon and garfinkle had this they had this song when I think back and all that crap I learned in high school that was Forest Hills High School oh that's amazing wow yeah and then you went from Forest Hills to Hamilton College correct well I would spent three years in the Navy and oh that's right yeah so uh um what what did you do while you were in the Navy well I volunteered to go into the Navy I

mean it unlike all the wars since it was a pretty black and white issue there and everybody hated Hitler and mhm and so all the kids my age B practically volunteered to go to and in in the Navy I was sent to this electronic technician school after boot camp they basically did a kind of filtering for performance ability then went through three of these Electronics Goes are more and more complex they took the top you know few from each school and at the end of that I was sent to MIT where they're developing early warning

radar system the reason they were developing is that they had these kamakazi planes that would fly just 20 ft above the surface of the water and we couldn't detect them until they were like 20 miles out and by then it was too late to get your your airplanes up to shoot them down MH these are they were Torpedo bombers they didn't have much flexibility just a thing carrying a big explosive and our fighter planes could easily shoot them down so somebody had the idea of putting a radar it was very primitive in those days early

early early days of Television a radar system in a plane and then you could scan the entire thing and this is all then sent by what would be ultra primitive television down to the ship uhhuh and I was responsible for the Electronics on the ship as it turned out and my buddy was responsible for the electronics in the in the plane it's amazing it was pretty exciting I know I I would have trouble with that responsibility now I have a whole naval squadron depending on you yeah but and it was 18-year-old kid but but it

illustrates it didn't at that age nothing bother you and it illustrates your uh quantitative nature and the um proclivity you have toward physics and Engineering did you study that at Hamilton I majored in physics and Mathematics at Hamilton College MH yes and did you enjoy your college Years yeah I I enjoyed them very much I was catching up so I only spent two years in h there were a lot of PE older older men and kids who took accelerated courses because we lost three years in the war so I you know went there when I

was 20 and took summer both years over took extra courses and uh graduated when I was 22 I see I I think I told you this but a number of years ago when I was bringing my children on their college interviews we went to Hamilton and I was thrilled to see in their new science building uh a display devoted to to you and your contributions as a famous alumnist of the college I'm sure you've seen that yes I it was there and I saw yeah yeah um and then from Hamilton you went to get a

PhD uh and I believe you started at Penn But somehow made your way to Hopkins uh could you tell us how that happened yes well at that time the well I switched for my interest in physics and Mathematics to bio physics medical physics and the reason for that was this like two years after the three years after the dropping of the atomic bombs on Hiroshima Nagasaki and I thought I don't want any talent I may have to be used for that sort of thing M and my roommate in College was a son of two Physicians

and they both told me about this and I was expressing my concern about going to that and they were telling me about this emerging field of biophysics and so I learned about it was quite excited about it and what they were doing is sending electrical properties of nerve cells which is where we started at the beginning of this thing everything was electricity old people working on the Brain they were electrophysiologist there' be biochemists using the brain for the research but it's only because of a very rich source of enzymes they were interested at all and

what was in there MH so any so you went to pan initially I went to pan and I got into biophysics there were two such departments in the country one was doing radioisotope Trace in at at Berkeley M and the other was his biophysics department at Penn which is Chaired by a guy named de L Bron uhuh who then moved one semester after I got there to John Hopkins to be president and he took with him one of the people a guy named keer Harline who won the Nobel Prize for his work on Vision he

became the chairman of the department and then then they moved a few of us with them from Penn to Hopkins so I spent the last four and a half years of my Graduate Studies at Hopkins and your PhD then was in uh biophysics biophysics yeah I was Studying the properties of ner cells as they degenerate mhm and then you mentioned earlier that you then uh spent a few years in England as a postto with whom did you work there I work with three people one was a man named Henry M who's best known for developing

M Chopper but he also this is a brain slicer where that uh was like a you know a deli slicer but to cut brains and slices and he made a couple of other Significant controvers mostly methodological and then from there I went to University of Cambridge or Cambridge University where I worked with a guy named EC Slater right after I got there he moved to Amsterdam so I spent a good part of that second year abroad working in Amsterdam M un oxidated [Music] phosphorilation uh which is his specialty I wanted don't I want to learn

More biochemistry because my degree had been almost exclusively in electrophysiology mhm and then after that I moved to the National Institute for medical research the reason for that is I wanted a department that had equipment facilities to do both biochemical and electrophysiological studies there was no place very unusual in those days for the bra the only place there was was a few pharmacology Laboratories which had bio some biochemical poy and that was the head of that group was man named vilhelm felberg MH who's very very important person in the history of pharmacology mhm he and

Henry Dale were kind of the Giants in neuropharmacology at that time in in England MH so let's finish if uh if you would just to give us a perspective of uh what what you see moving forward uh what you see in terms of how some of this this Many discoveries that your lab has made over the last several decades uh eventually influencing medicine you alluded to some interest today actually directly work done directly by your lab in the area of Alzheimer's disease and depression U more broadly many of us uh talk now about the difficulty

that the field has had in introducing new medications for neurologic and psychiatric diseases for half a century they've been no drug Right so I was wondering what your perspective is on that well it's clear and you know this very well Eric the reason why the same old drugs keep appearing is because they figured out to some extent how these drugs were mhm and so they keep using the same targets to develop better drugs and there's been substantial success less toxicity more rapid action longer onset longer uh duration of action and so on but basically nothing

important Conceptually now that we've learned all these things about signaling Pathways I plan to U spend uh whatever number of years I have left working M trying to uh do what's called today translation research to understand how our knowledge of these signaling Pathways can contribute to understanding diseases and the drugs that treat those diseases right and you gave us some examples earlier uh in Alzheimer's Disease and depression but also more broadly as you mentioned in Parkinson's disease and schizophrenia another I'm sorry another example is Parkinson's Disease Nat Hines and I uh with a extremely talented

postar fellow in our laboratory Miriam hymon in my laboratory developed this trap technology where it's now possible to look at all of the proteins being expressed in every individual nerve cell type in the brain uh the transcriptome if you Will and what we're doing with it now as one example in the case of Parkinson's it's known that in Parkinson's disease the there were two types of dopamine producing ner cells one group in the substantial and another group in the ventral te mental area which your anatomically very close to each other and we're now analyzing all

the proteins expressed in the two types of nerv cells mhm uh and we're finding large numbers of Differences between them and why this is exciting and potentially of great therapeutic interest is we're hoping that we can find cells in the uh substantial nagra that are very abundant there and they take identify those and then put them into VTA cells in numis and show that you can cause the death of those mice those of the doing cells those mice and conversely in much more practically importance take proteins which are very abundant in the VTA and Make

mice where you put them into the susceptible cells in the m and see if we can protect those uh cells from neurod degeneration mhm and that that technique is an example of how these modern molecular biology techniques can be enormously helpful in understanding how the brain works it really transformed the field of Neuroscience