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sciatrixI'm setting up my committee meeting arguments this weekend, which involve a plan to basically take my previous work (leptin injections in singing mice: increase singing behavior, possibly not song quality??? and also it turns out there's a big social order effect) and replicate it in a more sensitive and sophisticated way.
I'm going to use automated recordings before and after stimuli and no repeated effects at all, and then look at the animals' brains after a genomic action potential has the possibility to form. (Basically, experiences and behaviors will change the transcription of a bunch of genes in the brain in response; there are a few markers of this happening we can look for, telling us where in the brain something interesting is happening and giving us a pretty good chance of finding out exactly what it is. It also happens on a slower time-frame than electrochemical neurons firing and is more specific than the changes in blood flow that fMRIs measure it, but it does mean that I need to take brains away from their ow.) Except I honestly hate immunohistochemical work--I love reading it, that stuff is just fine, but it's not where my actual passions are, and what I would have wanted to do is check the RNA transcription in those parts of the brain but I straight up don't have time before I need to graduate. So I've passed that part of this study off to a postdoc to do in collaboration with me, and I'm intending to flesh out the non-brain part of this experiment to be my chapter 2. I'm looking at leptin-manipulated animals, same as before, and I'm looking at social manipulations, but I don't want to limit this work to just replicating chapter 1 minus the order effects.
So I'm thinking about how I want to present this study without the brain aspect to my committee. Thing is, I'm not just taking brains; I'm taking testes and the epididymal fat pad and livers and, most importantly, blood. I have blood plasma, even if it's just one time point. I can use that blood for things. Er, the epididymal fat pad is the one that wraps around the testis; in addition to being a decent proxy (I hope) for other fat pads, I found out last night that its presence is absolutely required for spermatogenesis to work. I don't think I'll bring that up with my committee other than checking for correlations within treatments with fat pad weights vs leptin levels, but it's certainly good to know and have.
One of the most important things about this presentation to my committee is going to be the choices I make about what we measure that blood plasma for, why I argue we should measure them, and what I expect to find. So I need to nail that down soon and get this presentation sanded down and ready for Wednesday. Trouble is, I'm still thinking.
The obvious markers to test are leptin (of course), so I can make sure the injections are actually working, and then things like insulin and free fatty acids and adiponectin: other metabolites so I can make sure that leptin is the thing that is really changing, probably, so that the thing I'm manipulating is what I think it is.
But I don't want to just test those things. I've been thinking about corticosterone and cortisol, and the growing evidence I keep finding in trauma and trauma-adjacent literature that leptin--which is, again, something most people think of as a satiety marker, something that is released by fat--also provides negative feedback for cort, and helps blunt stress responses and create cort sensitivity.
Trouble is, I'm not traumatizing these mice. They're getting one highly stressful but short experience (being injected with either control saline or leptin), half an hour, an hour of listening to something which might be a moderate stressor or might not be (as we've never tested this in either the tone or song conditions), and then an hour to chill at which point I'm taking blood. So do I have a timescale in which to measure anything? There's a single old study that did something similar, but they used a much longer and more severe stressor and a higher dose of leptin, too (about 4x mine).
It makes me think of the thing I think it wasrecessional said once, about how it took us so long to notice PTSD and trauma were even a Thing, because that was just How People Were. And pulling that back out to animals and animal social behavior is... almost a step towards what I want to be doing, if I can. First, this.
Also, probably relatedly: I am not sleeping for more than five hours or so at a time, and it is not getting better, and I am really fucking tired of falling asleep over my work, dammit. I had a rough meeting with my boss on Wednesday in which he basically gave me a hard time limit on graduating, and I need to turn on the skill, and I'm still broken but I've held it together this far on sheer cussedness and I'm not going to stop now. I want to look at this fucking thing. I want to look at fat and trauma and this weird little peptide that is doing way more than it gets credit for, and I have enough cussedness to see this damn thing through.
I did get in to see a new therapist, one with trauma training, yesterday for intake, on the theory that maybe change will help. I just about had left this post finished at that when my house started flooding, again, and I've just spent the last hour or so frantically bailing out the hallway and trying to make some kind of barrier against the rain. Thank fuck we have a Hoover mechanical mop... thing; we combined that with the Little Green, made a barrier of towels, and then K and I bailed like motherfuckers and T heroically ran into the side yard and set up emergency flood barriers. Thank fuck they're a paranoid genius and we had them on hand.
And my PI told me I needed to graduate by the spring or it wouldn't happen on Wednesday, and this meeting is... this Wednesday.
*hollow laughter* It never rains but it pours, huh?
I'm going to use automated recordings before and after stimuli and no repeated effects at all, and then look at the animals' brains after a genomic action potential has the possibility to form. (Basically, experiences and behaviors will change the transcription of a bunch of genes in the brain in response; there are a few markers of this happening we can look for, telling us where in the brain something interesting is happening and giving us a pretty good chance of finding out exactly what it is. It also happens on a slower time-frame than electrochemical neurons firing and is more specific than the changes in blood flow that fMRIs measure it, but it does mean that I need to take brains away from their ow.) Except I honestly hate immunohistochemical work--I love reading it, that stuff is just fine, but it's not where my actual passions are, and what I would have wanted to do is check the RNA transcription in those parts of the brain but I straight up don't have time before I need to graduate. So I've passed that part of this study off to a postdoc to do in collaboration with me, and I'm intending to flesh out the non-brain part of this experiment to be my chapter 2. I'm looking at leptin-manipulated animals, same as before, and I'm looking at social manipulations, but I don't want to limit this work to just replicating chapter 1 minus the order effects.
So I'm thinking about how I want to present this study without the brain aspect to my committee. Thing is, I'm not just taking brains; I'm taking testes and the epididymal fat pad and livers and, most importantly, blood. I have blood plasma, even if it's just one time point. I can use that blood for things. Er, the epididymal fat pad is the one that wraps around the testis; in addition to being a decent proxy (I hope) for other fat pads, I found out last night that its presence is absolutely required for spermatogenesis to work. I don't think I'll bring that up with my committee other than checking for correlations within treatments with fat pad weights vs leptin levels, but it's certainly good to know and have.
One of the most important things about this presentation to my committee is going to be the choices I make about what we measure that blood plasma for, why I argue we should measure them, and what I expect to find. So I need to nail that down soon and get this presentation sanded down and ready for Wednesday. Trouble is, I'm still thinking.
The obvious markers to test are leptin (of course), so I can make sure the injections are actually working, and then things like insulin and free fatty acids and adiponectin: other metabolites so I can make sure that leptin is the thing that is really changing, probably, so that the thing I'm manipulating is what I think it is.
But I don't want to just test those things. I've been thinking about corticosterone and cortisol, and the growing evidence I keep finding in trauma and trauma-adjacent literature that leptin--which is, again, something most people think of as a satiety marker, something that is released by fat--also provides negative feedback for cort, and helps blunt stress responses and create cort sensitivity.
Trouble is, I'm not traumatizing these mice. They're getting one highly stressful but short experience (being injected with either control saline or leptin), half an hour, an hour of listening to something which might be a moderate stressor or might not be (as we've never tested this in either the tone or song conditions), and then an hour to chill at which point I'm taking blood. So do I have a timescale in which to measure anything? There's a single old study that did something similar, but they used a much longer and more severe stressor and a higher dose of leptin, too (about 4x mine).
It makes me think of the thing I think it was
recessional said once, about how it took us so long to notice PTSD and trauma were even a Thing, because that was just How People Were. And pulling that back out to animals and animal social behavior is... almost a step towards what I want to be doing, if I can. First, this.Also, probably relatedly: I am not sleeping for more than five hours or so at a time, and it is not getting better, and I am really fucking tired of falling asleep over my work, dammit. I had a rough meeting with my boss on Wednesday in which he basically gave me a hard time limit on graduating, and I need to turn on the skill, and I'm still broken but I've held it together this far on sheer cussedness and I'm not going to stop now. I want to look at this fucking thing. I want to look at fat and trauma and this weird little peptide that is doing way more than it gets credit for, and I have enough cussedness to see this damn thing through.
I did get in to see a new therapist, one with trauma training, yesterday for intake, on the theory that maybe change will help. I just about had left this post finished at that when my house started flooding, again, and I've just spent the last hour or so frantically bailing out the hallway and trying to make some kind of barrier against the rain. Thank fuck we have a Hoover mechanical mop... thing; we combined that with the Little Green, made a barrier of towels, and then K and I bailed like motherfuckers and T heroically ran into the side yard and set up emergency flood barriers. Thank fuck they're a paranoid genius and we had them on hand.
And my PI told me I needed to graduate by the spring or it wouldn't happen on Wednesday, and this meeting is... this Wednesday.
*hollow laughter* It never rains but it pours, huh?
no subject
Date: 2019-05-06 08:45 pm (UTC)I've been working either on de-flooding my house or this presentation nonstop for going on five days and I swear to god on Friday afternoon (when I give the version that is Presentation for Seminar) I'm legitimately going to fall over and sleep for a week. Assuming that will be possible given the sheer level of humidifiers now acting in my goddamn house.