 This image does not do justice to the final product of the confocal microscopy; the result is brilliant overlay of the YFP-labeled and c-fos-labeled cells. What one is looking at are the traces of memory in this particular section of the brain, which is extraordinary. A steady tide of words flowed out in rapid progression. I spoke quickly, my hands flitting about as I attempted to describe the figures projected on the screen. Christine, my PI, stopped me politely and asked me to slow down, prompting me to attempt a more careful analysis of the graphs. It was the first time that I had presented on a scientific paper, and though the audience was small, the modest confidence I usually have while public speaking wavered a bit, given the topic. Condensing a dense Nature paper with a number of supplementary figures is a hefty feat, I discovered to my chagrin. It was a lesson in what I’d always said I’d wished to do: relay the jargon of science into understandable terms. Though I did not speak with as much grace and fluency as I had hoped, I was glad to have taken a step towards this goal. I was able to discuss with Christine what I did right and what I could improve upon, and I was renewed with a sense that I can tackle another presentation in the near future, but this time in front of the entire Hen lab.
Following conversation and planning comes the actual execution of an experiment, conducted in scrupulous detail. My days are often blocked by a number of different behavioral experiments to run with the mice allotted to the several projects I am currently a part of. Most notably, I have finally gotten up and running in experimenting with my Nestin mice that have the NR2B subunit of the NMDA receptor knocked out. I administered ketamine to these mice and am determining whether the NR2B subunit is necessary for the effects of ketamine. The first experiment I ran had a highly significant effect; the feeling of realizing that the data reveals a huge effect is like none other. I am continuing with this cohort to determine whether the result is even more robust with a higher sample size. In another experiment, conducted just today, I have figured out that the effects of ketamine have a prophylactic, or preventative, effect against stress. Seeing the results beautifully graphed on statistical analysis software caused my PI and I to squeal with joy. When I am not running experiments, I am able to improve other techniques such as perfusions, brain slicing, mounting, and confocal microscopy. Usually, when someone begins to conduct something that others in the lab have not yet experienced, time is allotted so that everyone can be present and learn as much as possible. I’ve found that this emphasis on keeping everyone on the same page allows us to have a stronger foundation; there is almost always backup if something goes wrong or if one needs help in finding or completing something. The world of research is so murky and dauntingly complex; yet, the spirit with which I’ve come to associate the pursuit for answers has gotten me hooked on the occupation. I am one who thrives on busyness, movement, constant inquiry and periodic reflection. Though I still cannot relinquish my penchant for leadership development and writing, I am finding it easier to find ways to intermingle my other interests with my central passion. My plan at the moment is to continue working in this lab through my college years, which includes completing my senior thesis. I am also beginning to look into grad school options and preparing myself to study for the GRE, which I am to take next year. It is both strange and exhilarating to begin thinking about how I can prepare myself to move forward professionally in the field of research. One day, while observing my daily, inextinguishable enthusiasm, my colleague quipped: “It looks like you’ve found your calling.” Though my current outlook may be colored by the bright glean of new ambition, I was struck by the truth of this casual statement. Isn’t this what people speak of finding: the job that doesn’t feel like work, the enthusiasm pervading one’s daily environment? This summer has gone by in a frenzy, with so much bombarded on me and yet so much knowledge and skill gained. It has, more notably, been a pleasure to have so much in common with whom those I work, whom poke fun at my clumsiness and can talk about restaurant week in between running around. This summer has summarily been defined by the daily succession of experiments and sometimes-unsuccessful scientific pursuits. Yet, I have found that I will most poignantly remember my summer days being invigorating, fun, and punctuated by the sparkle and buoyancy of laughter.  In this photo, I am using the confocal microscope in order to image slides that have brain slices on them. Since I’ve started working here, I’ve been slowly gaining the trust and confidence of my Principal Investigator, Christine Denny, and am now juggling roughly 4 projects. Over the course of the past few weeks, I have gained a number of technical skills, including intraperitoneal (IP) injection of mice; what’s more, I learned to do so on notoriously fidgety C57BL/6 mice. I was uncertain and fumbling at first, but with the aid of my colleagues, I am now teaching others in the lab how to make a drug and administer an IP injection. I have transitioned the responsibility of taking care of all of the breeding lines (as well as their genotyping) in the animal room to the technician. This gives me more time to focus on my behavioral experiments, perfusions, surgery, etc., and finally putting my data together.
The aforementioned four projects all span slightly different realms of neuroscience that all interest me tremendously. The first is one that I had been working on since I first started in January, which looks at pattern separation and how it differs in the brain from 6 month old, 1 year old, or 2 year old mice. The last time I looked at the data, the difference between the groups in the number of neurons in a particular brain region in the hippocampus was highly significant. Another two projects involve ketamine, a fast-acting antidepressant that we are using to determine its prophylactic effects to prevent the onset of stress disorders (such as PTSD in humans). I am working with Rebecca to refine her project to be re-submitted to the journal she originally submitted to, but the other project is completely mine, which is asking whether the NR2B subunit of the NMDA receptor in the brain is necessary for ketamine to work as an antidepressant. The final project is one I am conducting with Jennifer, the post-doc, which utilizes our AD mouse model. Since we are piloting studies, we are running a number of behavioral tests to get a baseline reading of anxiety and depressive-like behavior in these mice. We are planning to test their short-term and long-term memory in a few weeks, and finally stain their brains for plaques and tangles in order to see where the memory deficits in their brains occur. The lab atmosphere invigorates the constant work being accomplished, which I mentioned in my previous report. A few weeks ago, a pivotal day for Christine came when an inspector from the NIH—which awarded her the prestigious Early Independence Award—came to purvey the lab. He was a curious man, asking all of the lab members a number of questions ranging from hard data to everyday tasks and overall impressions. Though Christine and those who had been in the lab longer than me were nervous to be the main representatives of the lab, they had no reason to be anxious, as the truth spoke for itself. I listened and chimed in as we praised Christine’s incredible organization, her intelligence and humor, as well as her encouragement of new project ideas from us undergraduates. We were able to speak knowledgeably about our projects, and how each component of the tasks for that day fit into the larger picture. As I look ahead to the coming weeks, I plan to continue to help Jennifer with conducting behavior on the AD mice to determine whether they demonstrate a deficit in anxiety or depressive-like behaviors. I also plan to finish up with imaging the slides for my pattern separation mice, and deciding whether to run another cohort to get even more robust, defined data. Of course, my main project involving the NR2B subunit and its importance in the effects of ketamine will still be moving swiftly forward. Most excitingly, I shall begin working with Rebecca to improve her data with more evidence of the resilience effects of ketamine, to be submitted to a major journal. We met today, and the prospects are exciting; I may, should I see these experiments through, become second author on the paper. Overall, I still receive praise for my work ethic and determination, but I still wish to improve, and have the sense that there is a great deal to learn. I see how successful an experiment can be, but also how it can go awry. I read the reviewer’s comments to Rebecca’s paper, and realized how methodical and holistic one must be to prove your data is significant. Each day, I remind myself that organization, careful precision, and keeping up with current information about this growing field are integral in ensuring that my ideas are innovative and fruitful. If there is one thing that absolutely amazes me the most each day about working in Christine Ann Denny’s neuroscience lab at the New York State Psychiatric Institute—in addition to the fascinating work I engage in—it’s how the hours seem to vanish, whiling away without my noticing until I am reminded by the light that the sun is setting. This fact in itself is not what elicits my amazement; it is what takes up those hours, and how I have busied myself into an invigorating frenzy of science and innovation that makes me excited to delve further into the field of neuroscience research.
Transitioning from working 12 hours per week during the school year to full-time 40 hours per week during the summer has been both challenging and thrilling. Throughout the semester, I relied on Christine and the lab’s technician, Nikki, for assistance and guidance. Nikki trained me on a number of microscopes, had taught me how to run PCRs and gel electrophoresis, and had even brought me through the elaborate strategic technicalities of breeding and caring for transgenic mice (of which Christine has developed roughly 12 lines) in the animal room. Often, I would go into the lab to see a list of tasks to accomplish. Yet, during the summer, I received the shocking knowledge of Nikki’s planned 3-week absence to Spain. Suddenly, I was to be in charge of keeping up with all of the lines in the animal room, and seeing each experiment through from start to finish, from breeding the mice to genotyping, performing behavioral tests, cutting the brains, conducting immunohistochemistry on the brain slices, and analyzing the data. Christine, of course, aids me along the way. But, I took it upon myself to learn about the best strategies for breeding, to pore over the protocols written by other lab members, and to check and double check all that I am doing (for, as a beginner, my susceptibility to mistakes is perilously high). It’s been a great, though quick-paced, few weeks. Soon, we are to start on optogenetics on our mice, a recently developed technique of flashing light on the brain to enhance or delete cells of interest. Behavioral tests and injections of tamoxifen are in line for these next couple of days. In short, there is plenty of work ahead of me, and I’m prepared to take on the challenge. |
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