00:00and it was at a time where I was told to my face, but also that was the atmosphere, that we don't need anatomy anymore because we have imaging. And this was a very frustrating thing for me. And I went to, I was giving a talk at Oxford, and there's Tim Behrens and Matthew Rushworth in the front row, and they're asking all these questions about it, and the paper had like literally just come out while I was on the plane going over, right? Like it just literally came out. I didn't know this would be really too long. Thanks. Welcome to Stimulating Brains. Stimulating Brains Stimulating Brains 01:29Stimulating Brains Stimulating Brains 02:29Stimulating Brains Stimulating Brains 03:29Stimulating Brains Stimulating Brains 04:29Stimulating Brains Stimulating Brains 05:29Stimulating Brains Stimulating Brains 06:29Stimulating Brains Stimulating Brains 07:29Stimulating Brains Stimulating Brains 08:29Stimulating Brains Stimulating Brains 09:29Stimulating Brains Stimulating Brains 10:29Stimulating Brains Stimulating Brains 11:29Stimulating Brains Stimulating Brains 12:29Stimulating Brains Really cool. So you mentioned the endocrine accumbens and also amphetamine, your first PhD work. And so it always seems to me you did some things in the motor system, but I think you're kind of drawn to the striatum, pallid, you know, psychiatric or associative limbic system. Yeah, I never really did that. I mean, it's the only reason that I'd be associated with the motor system is because the basal ganglia is above. 13:02But I always was interested in the non-motor part of it. The limbic and the tibial. Why is that? Was there any... Yeah, because it goes right back to what I was drawn to in the first place, mental health. Mental health, okay. And how you explain things, which is where I'm still, many, many years later. How do you explain that people, on the one hand, rationally know that drugs are not good for them, or that in OCD your ritual is not going to be productive, but you can't stop doing it. Mm-hmm. So that kind of thing has always been very intriguing to me. Yeah, yeah. Sounds great. So a big jump in time, but we'll also go back in time again. But your more recent work, I think one key thing that... There are so many things that make your work interesting, but I think for the imaging field what makes your work interesting is that you're one of the few people that actually compare tracer with non-invasive imaging, 14:06like tractography, diffusion-based tractography study. And I think one of your key partners in crime, there are many others, but it's Anastasia Yandiki, who is the author of the Tracula software, which is part of FreeSurfer, and she's also doing a lot of other things often with you. So together, I think the two of you, probably with others, you also set up the Iron Track Challenge to look at that, right? Can you tell us a bit what that is? Yeah, so again, if you don't mind me going back in history completely, cut all of that out. I had decided to spend a little bit of time at Mass General, and I wanted to see what was going on at MGH. And so it got arranged by various people I knew and so on, and then I would come and have a visiting appointment there. And one of the things that I was always very interested in is circuitry. 15:02You know, if I felt like, okay, if we understood how the brain was, why we were doing this, why we were doing that, why we were wired together, we would really understand disease. Of course, that was naive, but I still believe that we'll, you know, have a better idea about how the brain is put together. And I had been frustrated, as many people are in basic science, when they see these imaging papers come out, and they make statements about circuitry or function or whatever. And it was at a time where I was told, I'm going to have to do a lot of research on this, and I'm going to have to do a lot of research on this, and I'm going to have to do a lot of research on this, and I'm going to have to do a lot of research on this, and we don't need anatomy anymore because we have imaging. And this was a very frustrating thing for me. And so I had many, many papers. I would get a paper with a new track, new pathway, and I'd say, oh, that's really pretty, it's very lovely, but actually that track doesn't exist. And, you know, it was frustrating. And so at one point I thought, like, okay, I'm not going to fight this. 16:04You know, diffusion is just... way, way too popular. And us anatomists, you know, like three of us in the world, maybe I'm just making that up. So I'm going to have to join them. So I had a paper, we had a paper called the Pathways Paper. It was with Lehman et al. It was the first paper, right? And I went to, I was giving a talk at Oxford, and I'm sitting there and I'm giving this talk, and there's... Tim Behrens and Matthew Rushworth in the front row, and they're asking all these questions about it, and the paper had, like, literally just come out while I was on the plane going over, right? Like, it just literally came out. And, or the day before, something like that. And I felt like, they know this paper way too well. They must have reviewed it. And so, and... 17:00Anyway, so we had lunch, and it was just wonderful because they were just so interested in trying to figure, figure out how Pathways really... And I was just so happy about that, that people really wanted to do that. So then I went to MGH, and Anastasia was another person like that. So I had interviewed, or interviewed, not interviewed, but, you know, set appointments with many, many people at MGH. And we just clicked, and, you know, she was really interested in it. And so it's been, you know, it's been a great journey with her, trying to... Exactly do that. So the whole goal is to take tracing and use the tracing as the so-called gold standard for understanding what tractography can and cannot tell us. And that was the origin for the iron track. So it's an open data set where people can test their diffusion tractography algorithms, and then they hand in solutions, and you... 18:00Right. So we first gave them... The first thing they got was actually the tracing. Okay. They got actually the tracing, and then the seed, and then they had to match the tracing with the seed. Okay. And they developed an algorithm to do that. Okay? So now they've got what they're supposed to match. Then the challenge, then, is to give them a seed somewhere else. Yeah. And the interesting thing about that was that they didn't do as well. They didn't do that well. Yeah. And the thing... I mean, the... Yeah. And so now she has a good... Yeah. Yeah. Yeah. Yeah. Which I'm part of also, which is asking the question, you know, it depends on where you're coming from. You know, it depends on where that seed is in the cortex. There are different rules in terms of how one gets to their terminal field, depending on what they have to navigate through. So now we're trying to do that from different perspectives. 19:02Sounds great. So I think I've heard you mention a few times in talks and different occasions that probabilistic tractography was superior to deterministic tractography as maybe one take home maybe to play devil's advocate I sometimes think that this choice and related choices could be seen as a typical bias variance problem you know you get more tracks and maybe also the right track with probabilistic but then would you also get more false positives so you know maybe it's more sensitive but would you also get more specificity yes yes yes so I I don't actually know the answer to that I think that that's really very very possible the reason we came to that conclusion is that there was a graduate student that we shared and that is also a published paper I think it's in 20:00there a woman yeah so in that paper what we did was we looked at all the different way we have this you know ground truth pathways yeah and then we have an animal that was of course that that animal had been scanned as well and tried all kinds of different systems on it and just found that the probabilistic work best was the one that came the closest with the most accurate you identification but it may be that in certain areas that's not true you know it really depends I think I think the thing that we found was that depending on where you are in the brain I mean one of the things that we thought about quite a lot is that maybe one of the mistakes we make in tractography is that we scan the individual equally everywhere in the brain and there's some places where the tractography has no trouble right it's just like a you know so makes sense 21:00to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to to brainstem or so yeah um cool so you've mentioned the Lehman paper that was I think 2011 in journal of neuroscience and there was maybe um a six uh second paper 2018 Safadi et al same journal that is right a bit in the same um concept where you looked at the a-league and related systems and the two across the two is it even possible to summarize some of the findings in a podcast like this yeah I think the main thing is that um fibers from cortex regardless of where they are 22:03um are going to enter the white matter in a certain way and that's that certain way is just because of the constraints of the brain and then they travel through the brain to their terminals and these an example is the alec the anterior limb of the internal capsule um and they organize themselves in the capsule and this is true for some other fiber bundles like the corpus callosum in a way that makes sense from where they're coming from and where they're going. And so the goal of those two papers really was to ask the question, are there rules that fibers use to get to where they're going that are going to be translatable into the human? The monkey brain is much smaller, much less evolved. Obviously, the cortex is tiny compared to the human brain. But some of the constraints are the same. The fibers aren't going to go through the ventricles. They're not going to go through parts of the striatum and so on and so forth. So they have to get to where they're going with similar constraints. 23:03And so I think the main thing about those papers was that there are logical rules that fibers use to get to where they're going. And moreover, within a pathway, within a bundle, the organization can be predicted based on where they are. So for example, in the ALEC, which I think was very helpful, if you live in the dorsal part of the cortex, your fibers will be dorsal to the ones that are in the ventral part of the cortex. It's a simple rule, but it helps organize how they are within the capsule. So I think both of those papers helped people look at these big white matter bundles and say, okay, there's an organization within it. Great. And then I guess a second or third really key paper, landmark paper for the DBS field is the 24:02Haynes and Haber STN paper that I think everybody in the field would know because it's really the gold standard, at least from modern times, I would say, where you parcellated the STN. And you did show segregated loops. So I also interviewed Malon Delon, on the podcast. And he mentioned that their basal ganglia circuit model was key to setting up these parallel loops in the first place. Did their work influence your thinking that may have led to this Haynes and Haber paper? Or was that important? Or was that a whole different time? Well, you know, it was influential because I didn't agree with it. And I still don't agree with it. I think that there's a general topography. That's definitely true. There's areas, you know, the areas that are really important. And I think that's really important. And I think that's really motor striatum and the motor, and then there's a limbic piece and a cognitive piece. 25:00All of that is true. They're general. But they're not strictly segregated. There's a huge amount of convergence. And that's true in the STN, too. So obviously, if you're in the anterior medial where you're stimulating for OCD, you're probably not going to be in the motor area. Yeah. But you're not going to be able to differentiate between anterior cingulate and dorsal medial prefrontal cortex. Those areas are just going to be converging in that region. And the same is true with the striatum. The idea that you've got a connection from dorsolateral prefrontal cortex that's not interacting with a connection from ventral lateral prefrontal cortex and striatum, I don't think it's going to be. Well, it isn't true. So there's just a huge amount of convergence. So I think you have both things happening. You have a general topography, but a huge amount of convergence. So both. Parallel circuits and funnel, right? I know you. I don't. I mean, I don't particularly like the idea of parallel circuits because there's no doubt 26:06that M1 fibers, where people do a lot of work, are in their own world. And the same is true with area 25. Okay. Right? So there are these polar areas of the striatum and the basal ganglia. Are the most segregated from other areas. So the shallow, the nucleus accumbens, and the really dorsolateral caudal part of the striatum. But in between, you just have... So actually, I think a classic paper that was done by Charlie Wilson in rodents, where he did the very heroic experiments of filling individual... Individual cells in the cortex and following the axons into the striatum. And what he found was that an individual cortical neuron innervates up to 17% of the striatum. 27:08Okay. So it arborizes huge. And each one of those collaterals only contacts very minimally each cell. So it kind of goes popping from one to another, and then there are collaterals. And if you look at his drawings... They're just very beautiful drawings. You see how much it spreads. It's a single cell. Wow. Yeah. So to me, that right away argues against parallel segregated processing. And I think where it's very well described as well, same thing as the dopaminergic neurons. They even span broader patches, right? Exactly. Where if their role would be to say, yes, do that again, in simple terms, they would say that to nearly the whole striatum, or a big part of the striatum. Exactly. Yeah. Exactly. Okay. Makes sense. That's interesting. So, I mean, it certainly, there is a gradient, right? I guess you would agree with that, right? 28:00But it would, there's a lot of convergence, and maybe that is even the function of the whole thing, right? Yeah. Well, I think it is. I think what we find also is that the, and I mean, everybody knows that the corticostriatal projection is very patchy, right? And those patches don't necessarily stay in the same general geographic region. Right. Yeah. Yeah. So, you'll find a patch that's like somewhere else, and you're thinking, well, what is it doing there? If it's really this gradient, then you would expect, you know, like a rainbow to keep changing evenly. But it doesn't, because you've got these patches innervating areas that you're really surprised that it is. So, the other thing is that within cortex, I can give you the example of anterior cingulate, which is a very large area. And we found that there's an area that is kind of hub-like within the anterior cingulate, and that area projects to the striatum in a way that gives you patches everywhere. 29:01Mm-hmm. Okay. So, not all cortical areas really are not the same in terms of how they innervate the striatum. Okay. They don't, you know, they're... Yeah. Yeah. Very interesting. I think I had a follow-up question on the STN. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. So, when you look at the STN work, because we talked about the Haynes and Haber paper, I read in some reports that the STN would receive input from almost the entire frontal cortex. But do you think that's true, first of all? And then probably there's more projection from some regions versus others, right? Because in your paper, I think you selected five or so, or six regions. Would you think the other... Is that a general rule that everything... No. No. No. No. No. No. No. No. No. No. No. No. No. No. in the frontal cortex projects to the STN? I think it does. I think what happens, and we showed in the paper to some extent, but I think that the medial part really merges 30:02with the lateral hypothalamus in a way. So a lot of the anterior cingulate and those in there. You showed that for the VMPFC and OFC projections that they are in this ventral cusp of the STN, right? Yeah. That you would still count as STN, if I understand correctly, or part of it. Yeah, well, the reason, I mean, it's a very nebulous area, and it's an area that had been described prior to us. DeGirian described this in the 19-oh-whatever, when he wrote his very famous atlases. And he also describes that that area should be concluded with the STN. And cellular-wise, it has this mix of things. The reason we included it is because the ventral pallidum projects there. I see. And we showed that in a previous paper in the monkey. 31:02So the ventral pallidum really has a very strong projection, which continues into that lateral, that little comb region of the lateral hypothalamus. Interesting. So, you know, it depends on what you want to call the basal ganglia, right? Yeah, yeah, yeah. Larry? Swanson, I think, has this concept where all of the cell nuclei would be called either striatum or pallidum. Yeah. You can divide them into these two functions, doing inhibition and disinhibition, I think. That's his, like, what do you think of that? Would you agree that maybe everything is, you know, every cell nucleus is kind of part of either being a pallidum or a striatum, or could that be a possibility? Well, I mean, I think that's going a little too far because the morphology of the cells are different. You know, if you go back to how you define a pallidal cell, you know, and a hypothalamic cell and, you know, whatever, 32:01you've got very different morphologies there. So I think in the Swanson work, the hypothalamus would be separate. So that would be also part of that canonical circuit. But all other cells, like, for example, lateral septum, I think, would be, lateral medial septum would be a pair of striatum and pallidum as well, to him, I think, in his 2000 paper or so. Yeah. We leave it at could be, I guess. Yeah. Okay. So, great. So one thing that struck us when we studied the Hains and Haber paper in our lab in the journal club was that, all of the projections are based, and of course, that's natural if you know about it, but based on a single macaque, right? I think you had 43 animals. Five of them had two injections, so a total of 48 injections. 33:00So I think then one of these looped often is from one animal, right? Like if you would, for example, the OFC projection, that was one animal. Is that correct or did I understand that wrong? So, yeah, so, right, so we inject an animal and then we trace it out. That's correct. But we have many injections in the OFC. I see. In different animals. So, you know, of course, we're illustrating it with one animal per area, but we have, well, I don't know how many we have now. I mean, at that time, but, I mean, we've got, I think it's something like 150 injections. Yeah. Yeah. Cortex. Yeah. So. So you would know from other that it would replicate. Oh, yeah, yeah, yeah. No, that's really not, that's really not an issue. I mean, if you, there's, there are many, many injections to show. 34:04The same thing. The same thing. Yeah. Very cool. So going back to the imaging world with Cameron McIntyre alongside three other famous anatomists together with you, such as Peter Strick, Jolene Smith, and Martin Poirot, I think, you worked on creating a holographic tractography atlas for the subthalamic and palatal region published in Neuron 2019. And the data we just talked about, I think both from the Haynes and Haber paper, but also from the Ehrlich papers, were really key for building that atlas. So it's an amazing data set, and I'm a huge fan of the effort. So first of all, any thoughts about that general process? Can you maybe describe a bit how you created that? It was great fun. Yeah. I mean, it was just, they had an amazing system there, this hologram. It's, you know, altered reality, so you can, you see what's going on around you, but you 35:03actually are, you know, in the middle of this brain that's in three dimension, and you see these fibers going through, and I think as, you know, most anatomists, they have this sort of 3D world in their head of where things are going, and very often it's very difficult to either draw those or explain those. Yeah. But you have this sense in your mind, and so having this hologram was really, really fun, because you would be in the brain, and you know, they'd have these fibers, and it was almost like, you know, an anatomist's dream. They'd say, no, no, no, let's take this one and move it over here, take this one and move it over there. And it was great, you know, we could just adjust things the way we see them under the microscope. Yeah. And so it was a wonderful thing to do. Was there a lot of discussion? Like, would the four anatomists agree, usually, or? Yes. No, I think, you know, no, I think we, there was a lot of discussion. 36:01Yeah. But not because people disagreed, but because we were just trying to get all of our perspectives in there. I think the reason that, I mean, the nice thing about this was that the four of us really had expertise in different things. Yeah. In different parts of, so we had a basal ganglia person, Andre, not Andre. Martin Paul. Martin. Yeah. Martin, yeah. You know, it was basal ganglia. Yolen Smith was basal ganglia, but slightly different area. Peter, cerebellum, and thalamus, and so on and so forth. And I was basal ganglia, but internal capsule and STM. And so. It was really nice because, because, yeah, I think, you know, we all shared the love for this, but slightly different areas of the brain. So it was good. Yeah. Really cool. And I think, I mean, the data set has been made openly available, so that's great. 37:02And I think it is one of the, you know, only, probably the only data sets that have some of the details that you wouldn't really see with tractography, such as, for example, the, just the hyperdirect axonal collaterals that you would. Rarely see or not, you know, you wouldn't be able to reconstruct with tractography. So it could be used for, and we have used it and Cameron has used it for, you know, DBS network mapping, deep brain stimulation network mapping. And in contrast to the diffusion based connectomes, which are full of both false positives and false negatives, this Atlas is likely free from false positives. If you're right, right. There's no wrong tract in there. Right. But it would still have many false negatives. And what I mean with that is, you know, just tracks. You didn't draw in, right. Oh yeah. No, absolutely. Of course, there's more going on, right. And it has gaps and so on. So I don't know. Do you have thoughts on that? Or do you see that building on top of the data set and filling in the gaps continuously 38:02could be a good idea? Or because at some point it's really very hard, even with, you know, things that you are doing with Anastasia, I think, to come to a similar level of detail and accuracy with. Empirical data. But we only have that one Atlas now, right. So it's not possible to. I mean, there are just, for example, if we're interested in the PPN to STN connections in freezing of gate, the tract is not in there. So we wouldn't be able to see it because it hasn't been drawn in, right. Or some other details maybe. Do you see any value in or is there even a plan to continue that work? Well, I think he's writing another. He's writing a renewal. So which we're all part of again. Amazing. I think that you're absolutely right. I mean, it's a very skeleton drawing. It's if you want to stimulate the alec, if you want to stimulate the STN, it doesn't have a lot of the 39:07thalamic fibers that are going through that area. I don't think it has the H fields. Yeah. And those are important, especially if you're going to be stimulating. It has the answer and the fasciculus lenticularis. Right. But yeah, no, I think I agree. Not all the details. It doesn't have all. Yeah. It doesn't have all of them in there. And then maybe, maybe final, final question on the STN. I also read that the granular insular cortex projects to the STN. But I've only read that in one source. So I wanted to ask your expert opinion. Do you? Do you think that's true? That is there a projection from insular to STN? I can answer that next week. Let's do that. Well, no, my guess is that it would. 40:00Yeah. But we just had a spectacular injection in the in the insular cortex. Okay. And so, you know, just literally hot off the press. And my guess is I haven't looked, but I can. We can follow up. Great. We could follow up on that and I can answer that. Super. Okay. So you have mentioned that it's, or sorry, let's, let's go into your work on OCD. So I think you have quite a bit of grants in OCD and also interest in OCD and not only OCD, but the whole psychiatric diseases that would be also stimulated with the brain stimulation. And I think there's a really important article, review article in BioPsych from you in 2020. Yeah. On OCD, where you anatomically describe the four targets that would be, have been used to treat OCD. Any take-home conclusions you want to highlight from that paper? Yeah. So in that paper, you know, we talked about the four, four, four sites. I think that there are, there are a couple other sites that I would add to that. 41:00But, you know, those were the Alley, the internal capsule, anterior limb of the internal capsule, the ventral striatum, which are very much associated with the OCD. And then the other site. And then the other site. They're very much the same site, really. The STN and a brain stem site, which was targeting the ventral tegmental area. And I think the main take-home message in terms of the stimulation for therapeutic approaches is that all four sites get similar, some similar package of fibers. Yeah. But having said that, they also might get some different things. Mainly where they're based on where the electrode is. The other main point of this was, so for example, the Alex site is likely to get all the descending and ascending cortical fibers. Yeah. Okay. And then the striatal site is going to get some of those because it's going to capture 42:00the internal capsule, but it also will get all the basal ganglia connections. The STN site, again, the hyperdirect pathway. And the palatal connections and the ventral tegmental area. A number of different fibers that are passing through there. The interesting thing about this is really what else is going through each of those sites. So we have our focus, we have our lamp post, and the keys we're looking underneath that are the orbital anterior cingulate fiber pathways in basal ganglia. That's what we're looking for. Yeah. So we're looking for that. But we're not paying attention to all the other things that are going through there. True. And that's particularly true in the STN and in the brainstem. There's just a lot of stuff flowing through there. Those are small areas with many, many different fiber bundles. I think in both of those, you're likely to be involving many brainstem regions that you may, in a more robust way than you will in the more anterior targets. 43:03And I think that where we can learn from these is that we're going to be looking at the whole process. And what we're going to learn from these is to identify for which population of patients. I mean, this is one of the things we're doing for a separate group, is to develop a database for DBS. Right? So that everybody that contributes will put in their electrodes, all of the criteria, et cetera, et cetera, outcomes and so on. And I think once we have a database where we have a population of patients, and we have a large population of patients, that you can divide into symptoms and outcomes, I think we'll get a better idea about what targets might be better for which populations. Yes. Yeah. So it could really be, one could postulate that the general Y box, so the core obsessive-compulsive symptoms might, or it seems like, might have a similar improvement in all of these targets, 44:02but then maybe some patients with more anxiety, more depression, more anxiety, more anxiety, depression might have a slightly different, like better improvement in this target versus cognitive flexibility issues could be targeted in a different one. Right? So I love the idea that to have overlapping networks that also have shared networks, right, but then differences. Right. That's really cool. So, yeah, that's already super, super nice. We've covered a lot. I wanted to ask one more general question about anatomy. Right. You mentioned it before that you were, you said you were told to your face that at the time people didn't need anatomists anymore, but they, because they had imaging and I'm totally with you that this is so wrong, but, and I always wondered about that because I was not around in that time where imaging came around and it's a bit like, you know, movie TV killed the radio star. 45:02People thought they would see everything in the MRI, but they would not. So then I think there was this trough of anatomy and it seems like now it's gaining momentum again. Would you see it the same way or? Oh yeah, no, absolutely. I think that, I think the sad part of that is that during that time we just lost a lot of trainees. They're just not, they're just not trained. And I think there is a very big upswing. There's a big upswing for a sub-pro. A sub-population of imagers, not everybody, but who are really interested in knowing what the anatomy underneath it is like. And are not as quick to say, well, we're in the human and so we're different and so therefore we're right because we're only in an animal. And I think that was a lot of what we had gotten before, which is, you know, we're in a human and so. 46:01But I mean, even post-mortem anatomists were not around anymore. Well, yeah. They haven't been around for a very long time. And you could say, for example, going back to deep brain stimulation, the beginning of that era and field, you know, in the maybe 60s, 70s in all these schools, there was usually an anatomist in the OR or at least part of the team, part of the planning. You know, people like Rolf Hassler, for example, in the Freiburg School was really important. He was a key decision maker, student of the focus, I think. To guide the surgeons and the neurologists, right? So and then imaging came around and people thought they would see everything themselves and wouldn't need the anatomist anymore. But I really think we lost so much, not only training, but also knowledge, right? All the details. Because if you just look at it in that course view of the MRI and back in the day, it was even worse than now. 47:00You know, you see just the rough shapes, right? Well, that's right. And I think that very often, you know, that is a loss because when you spend a lot of time on the microscope and looking at it the way the old anatomist did, you can look at these images and you can see where the structures are. And I've gotten that many times where people say, how do you know it's there? And you know it's there because you've been, you know what it looks like when it emerges, when it doesn't, you know the structures around it and so on. So you can. Yeah. Yeah. And I think that that part, you know, might be coming back. The people that come to my lab to train are now mostly coming from imaging. Okay. Interesting. So they're mostly coming from a place that has done imaging and they want to know the anatomy underneath that. Okay. So just they're not too many of our labs around anymore, but I think that many imagers are now doing that. 48:00Yeah. Who are the other ones that come to mind? So we had Katrin Amunds on the podcast as well, who has the big brain data set in Jülich in Germany. And then I know a young colleague, Annika Alkemade, she's working with Berthe Forstmann. Right. And I would consider her an anatomist, even though she's young. Right. Right. Of course. Do you have other people that still do that, where people could train? You know, they're mostly in Europe. Okay. Interesting. Yeah. They're mostly in Europe. Helen Barbus is still quite active here. She does a little bit of different stuff, but still, I mean, she has a very, very strong anatomy background. Jeremy Schwaman is here still. You know, there are still a number of people that have that background. I guess the people that were in that McIntyre project would also. Oh, yeah. Of course. Of course. 49:00Right. Right. So, I'm not sure if you have any other questions. No, I'm good. Super. So, and then I guess at Rochester, you have amassed a vast database of climate tracing stains. You mentioned a few of them. And if I remember correctly, I might be wrong, but you were, you told me you were in the process of digitizing some of that. Where does that stand? Yeah, so that, we've had so much trouble with microscopes and, oh my God, I don't even want to go down there. Yeah. But despite that, we have really made some progress. And we have, I don't remember how many cases, we must have 20 or so, I mean, out of the hundreds, not that many it sounds like, but that are digitized for dark field mainly, some for bright field and a few for the nistle. We're kind of plugging through it. I think we're almost ready to start making that publicly available. Good. I'm just trying to figure out the best, you know, platform to do that on, but I think we're getting quite close to that. Amazing. 50:00Because I think at some point we had a joint lab meeting together also with the New York groups and you showed some of these projections and of course some are published as well. And I remember one so vividly was a projection site, I don't even know from where, but it was to the thalamus and it really flashed out these bright dots that were scattered within the thalamus. And I just realized we would never see that with imaging. We wouldn't see it in that way. It would always diffuse, you know, not in that crystallized and clear cut way. So it would be an amazing resource to have. So the thing that gets lost in imaging is that I think people assume, right, that these areas which are very large are kind of one thing. So you have dorsolateral prefrontal cortex or area 46 if you want to use Brodnum's areas. They're all very large. And you have this idea that A goes to B. Yeah. And B goes to C, et cetera, et cetera. 51:01But actually if you look at the anatomy when you look at projections, what you really see is this region, this very large region, and oh, there's the projection and here's the projection but nothing in between. So you've got these very complicated circuitry networks that are going on that you'll, it's hard to see in imaging. On the other hand, what we have found is that when you're looking at seed-based, you're looking at seed-based functional, so just going to seed-based, both functional and diffusion, if you know what you're looking for because you know what the anatomy should be, it can be remarkably accurate. And the strongest functional connections, I mean others have shown this and we're showing it now too, the strongest functional connection from resting state are actually ones that are hardwired. Okay. So the anatomy, yeah. And actually these are the strongest, yeah. 52:00Makes sense. So it does make sense but, you know, it's helpful because very often people will say, well, you know, it doesn't have to be directly connected. And even though that's technically true, you don't want to use that as an excuse, right? Yeah, true. You want to be able to say, well, these are and this isn't and so it's not, so why isn't it? Yeah. How is it getting there? Makes sense. So, yeah. Okay. So we've got a lot of your time already. So to wrap up, maybe just some rapid fire questions. First one, did you ever have true Eureka moments in your career where you thought, oh, now I get it or this was great or wins in science, yeah, successes? Well, I think the one that probably might stand out, it's also I think one of our most cited papers is the spiral. The spiral paper with the cortical, the striatal, striatal, nigrostriatal connection where we 53:06showed that the ventral striatum projects, there's a reciprocal connection but there's a nonreciprocal component to it. The nonreciprocal component projects to a more dorsal area. That area has a reciprocal but a nonreciprocal and it keeps kind of winding its way through as a way in which the emotion can impact. It can impact on cognition and motor systems. So yeah, I think that maybe that's a really cool paper. Yeah. Yeah. I think that would probably, and well, I guess the other thing is that fibers are organized in really clear ways. You know, the rules that we had mentioned. Okay. So, okay. That makes sense. Great. And then the opposite, any time you thought this was a waste of my time or this was a failure or this was, I don't know, not great. Yeah. Yeah. I have to think about that. 54:02I'm sure there are many of them. I've suppressed them all. Yeah. That's why I asked that. I think the thing though that, well, I think what I would say is that if I had to say that in a career way to people who are developing their career and you want to say, you know, you keep going on and blah, blah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. Yeah. at that time the fashion was to do EM and understand really whether this receptor was here 55:02and this general tracing was just not really important anymore. This is before imaging. And, you know, blank out said you should change and do EM. I said I'm not an EM person. I'm not going to do it. And this is what I want to do and this is what I'm going to do. And I got through that. And then the other is that, you know, when imaging came out, it says, you know, you're done. You're never going to do this, you know. You know, neuron, I will tell you, neuron, nature neuroscience still will not take any of my papers because they're descriptive. And I've been told that. I see. Okay. However, they do ask me to write reviews. They won't take the, you know, I've. You put a lot of papers. And they'll say, I don't do it anymore, of course. But they'll say it's too descriptive. We know it's important. It's a very important, very, very interesting point you're raising. 56:02I think in the foreword of the amazing book called Brains Through Time by Streeter, it's about anatomy, but, you know, how they develop over time phylogenetically. They have a whole section about exactly that, descriptive versus hypothesis-driven science. And it was the first time. I really realized this, you know, these two things. And they made the point that the whole field of astronomy would be descriptive, right? You can experiment on a star. There's a lot of these that our current focus on these hypothesis-driven things can be detrimental to the field as well. If we only allow that. But it's more than just that. If it was just that, maybe. But, you know, a lot of the knockouts and the knock-in animals are just purely descriptive. They don't know what's going to happen. They're just like, oh, dopamine receptor, that's really cool. Let's knock it out and see what happens. That's descriptive. Sure. But that's accepted. 57:00I see. So it's not that kind of, you know, we need good description in order to develop a hypothesis. So I've learned, you know, you have to figure out how to work the system. I've learned to make a hypothesis out of something that isn't a hypothesis. I hypothesize that area 24 is going to project here. And I'm like, okay, I know a hypothesis, you know. Yeah, it makes sense. And, you know, like, okay, it's part of what people want. But the reality is, if you look at really good science, the only thing, the thing that comes before the hypothesis is really good observation. Absolutely. Yeah. I couldn't agree more. So you mentioned advice for young researchers entering neuroscience. So be persistent is maybe one thing. Do you have other advice? Yeah. 58:00I think the other thing I would say is that there's a lot of interesting things in science, a lot of interesting directions to go. But you have to be, you have to end up doing something that you will enjoy doing every day. So every science is very tedious. There's tedium in everything. And I remember doing electrophysiology. I went to a Cold Spring Harbor course. I thought it was amazing. It was so cool. But I knew I could never do it. Because fiddling with those rigs was just not something I wanted to spend my time doing. Whereas looking under the microscope was. And so I think that finding something that you really not only intellectually love, but fits your personality and what you, you know, how you like to organize your day, et cetera, et cetera. I think is really important. And the persistent stuff, I think is, you know, it's this cliché, follow your heart, which I hate. 59:01Yeah. Like anybody else. But your heart has, you know, you have to have some fundamental reason for that. Sure. And when you do, I think that you really need to push through. Makes sense. All the negative things you'll get. Any advice for women in the field? Yeah. Well, it's the same advice. Okay. Yeah. Yeah. I think don't, you know, it's the same thing everyone always says. Don't punish yourself for your personality. Sometimes you're just not an assertive person. That's okay. I think that, you know, there's a lot of push to be assertive and to be like, you know, pushing yourself and so on. But, you know, you have to accept your personality too. I think that this, you have to kind of balance that. Accept your personality and also try to move outside of it a little bit. Yeah. I think there's a lot of frustration with that. Yeah. Women who are quiet and maybe more reserved and that's the way they were brought up. Yeah. Or maybe that's their personality. Yeah. So. Makes sense. What do you think the future of the field will look like? 01:00:03Anatomy or neuroscience, neuroimaging, whatever you want to talk about. Maybe anatomy is the most interesting one. Well, I still think that there's so much to learn in the basic anatomy. Some of the techniques that are coming out now are fantastic like these. Right. You know. You really can see, you know, so we have some pathways and so we have to laboriously carry it out, chart it out. But you take this whole slab and you can see the fibers just going through the slab. It's just really nice. Nice. I'm not sure that's what your question was. What was your question? Yeah. I mean, maybe if you were to paint a broader picture, what will we do in 20 years? You know, what is the future of anatomy going to look like? Is it going to be? I don't know, simulations or new techniques? It's hard to forecast these things, I know. But you know, what would be your dream to have in 20 years? 01:01:01Well, I think a network, to be able to really visualize a network in 3D with all of the components to it. Yeah. Yeah. I mean, I think that would be. Be great. Okay. Any missed opportunities we are currently having in the field? Things we should be doing but are not doing enough? I guess an anatomist one. Well, in a general way, I think science is just too technique oriented. Too new tool development oriented. And there's not the intellectual. I think there's not as much intellectual. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. Okay. 01:02:06the day had the luxury to do that. So why would you not have that time now? Just because of the pace of things? Well, the pace of things, yeah. I mean, you have to, you know, first of all, you know, administrative things have just gotten kind of ridiculously complicated. There are lots of rules and regulations and so as a senior investigator, as part, you know, as the head of the Conte, there's such a huge amount of time that is devoted to just stuff that, you know, often is silly in my opinion, but, and everybody's opinion, not just mine. And then there's a lot of pressure to publish, from a scientist's perspective, there's a lot of pressure to publish in these high-end journals with lots and lots of different data. All has to be very cool, new tools. The fact is that a lot of the old tools, you know, 01:03:01still... I mean, traceable. Yeah. ...and the processing is just great. Yeah. I mean, you know, I use methods that are 40 years old and we're still getting very good data and good papers and... Yeah. ...but there's a lot of pressure for people to move faster with new techniques that they don't really know how to use. So yeah, anything we did not talk about, I know we covered a lot and I took a lot of your time, but any question you would have left me to ask or that we did not talk about? No. No? No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. 01:04:00No. No. Thank you.