Download Previous Research Essay 2012 - University of Cincinnati Graduate

Previous Research Essay 2012 –Ryan Makinson
My decision to enter into the field of neuroscience and pursue a Ph.D. at the University of
Cincinnati is related to my lab experiences as an undergrad at Emory University. During my
freshman year at Emory, I had the opportunity to work in a laboratory at the Center for Disease
Control (CDC), where I was given a position as a general lab assistant. This job provided
excellent experiences that introduced me to the laboratory environment and scientific process.
Working with a team of scientists, the primary focus of this research was to investigate the
biological mechanisms underlying venereal diseases. Specifically, we were interested in
developing novel blood tests for the detection of syphilis. This experience opened my eyes to the
importance of such research, and its impact on human health.
The following year, I was accepted into Emory’s SIRE research partner program, which
facilitated a unique opportunity to study depression under Dr. Charles Nemeroff. One of the
overarching aims of Dr. Nemeroff’s research was to investigate the CRH system as a potential
therapeutic target for antidepressants. The research project I was involved with included the
study of CRH expression in the brain, which, in elevated levels, is associated with major
depressive disorder (MDD) and anxiety. My project entailed examining the expression patterns
of CRH and other stress-related genes in relevant brain regions, including the hypothalamus and
amygdala. As part of this study, rats were injected with a lentiviral construct to induce chronic
overexpression of CRH in the central amygdala (CeA) followed by behavioral tests and tissue
collection. Cryostat sectioning of the amygdala and hypothalamus was conducted before in situ
Hybridization and tissue analysis. The results from this investigation supported our hypothesis
that, following lentivirus-mediated CRH overexpression in the CeA, an increase in CRH
facilitated HPA axis hyperactivity would be observed. Correspondingly, BDNF increased in the
PVN while BDNF decreased in the DG. These results further clarified regional and quantitative
differences in expression patterns of CRH and BDNF within the brain and pointed to the
hippocampus and amygdala as critical regions for a new class of antidepressants that act on the
CRH system. The technical skills I gained in this lab (such as the use of lentiviral constructs) will
be directly pertinent to the completion of my proposed GRFP research project.
The following summer, between my sophomore and junior year, I was again fortunate to
be accepted into Emory’s IRES program, which supports undergraduate research abroad. During
this time, I conducted my own project in immunology under Dr. Patrick Bertolino at the
University of Sydney. This project involved investigating the interactions between the liver and
the immune system in order to understand the tolerogenic property of this organ. With my
project, I demonstrated that treating donor T-cells with pertussis toxin can be used as a means of
diverting T-cell tracking to the liver. Previous research projects in this lab have shown that once
naïve T-cells enter into the liver, T-cell activation is specific to this organ and therfore presents a
possible mechanism to facilitate the process of organ transplantation. The time abroad also
showed me how scientific research is often an international process; I was supprised to learn
when I arrived in Austrialia that my PI was familiar with many projects and faculty members at
Emory University. Overall, I found spending time overseas was a highly enriching experience.
Upon my return to Emory, I sought to expand my training by approaching affective
disorders using a completely different technique and animal model. Dr. Mark Wilson and Dr.
Mar Sanchez at the Yerkes National Primate Research Center use behavioral testing and
observation, biochemistry, and genetics to study prosocial and anxiety behavior in non-human
primates. This research position was also made possible in part by the HHMI Neuroscience
Initiative Scholar of Emory fellowship I received during my junior year. In this project, I work
with a team of researchers to study the combined effects of social subordination stress, genetics
and developmental exposure to estradiol on behavior and emotional development in female
adolescent rhesus monkeys. Specifically, I conducted, recorded, and analyzed behavioral tests
designed to assess emotional reactivity to mildly stressful or threatening situations. We
hypothesize that chronic stress, imposed by social subordination, in combination with the short
variant of the serotonin reuptake transporter (SERT) gene will increase emotional reactivity to
novel situations. This longitudinal investigation is still ongoing, but results obtained thus far
support our primary hypothesis. In collaboration with other projects in Dr. Wilson’s lab, this
interdisciplinary study aims to illuminate some of the complex environmental and biological
interactions that contribute to the pathology and expression of anxiety-related disorders and other
common psychiatric illnesses. Dr. Wilson, Dr. Mar Sanchez and I are have recently submitted a
review on the role of estradiol on affect and psychopathology during adolescence. From my
work with Dr. Wilson and Dr. Nemeroff, I have presented four poster presentations.
Following graduation, my personal interest in affective disorders led me to the University
of North Carolina at Greensboro where I worked as a research assistant with Dr. Kari Eddington
and the Depression Treatment and Research Program studying the cognitive and motivational
processes in adult depression. I also worked on a neurofeedback brain-mapping project using
Quantitative Electroencephalography (qEEG). From these projects, I greatly appreciate the
clinical-based research experience. Overall, my curiosity in neuroscience is driven by a desire to
understand the biological mechanisms of the brain, but this lab has allowed me to see the
psychological disorders I have studied from both an intellectual and personal standpoint.
Currently, my research education has continued in my graduate work at the U. of
Cincinnati in the School of Medicine where I am participating in my first lab rotation. Under Dr.
James Herman, I am looking at the expression of the glucocorticoid receptor (GR) in the
prefrontal cortex (PFC). So far, we have been successful in isolating and extracting the GR
protein. Next, we will be working on quantifying the expression of the GR in stressed and nonstressed rats. This experience has inspired me to submit this NSF proposal, and this work will be
incorporated as part of my larger GRFP research project proposal. In relation to my proposal,
this lab has considerable success in using viral vector constructs for gene knockdown and
overexpression. Importantly, my work in this rotation has helped prepared me to successfully
undertake my proposed GRFP research project by learning new skill sets and optimizing
research protocols. As with any ambitious research objective, I anticipate potential setbacks and
technical challenges, but with the support of my lab members, I am quickly learning how to
successfully troubleshoot when problems do arise. With this in mind, I am fully confident that I
can effectively carryout my GRFP research project with the eventual goal of presenting this
research in the form of papers and poster presentations. I also plan on having undergrad and high
school students participate in all aspects of this project.
Together, my previous research experiences have span intellectually broad topics
covering multiple disciplines. I have enjoyed the opportunity to design experiments, analyze
results, present findings and publish papers. But most importantly, I feel that I have been
involved in projects that are meaningful and have the possibility to make significant
contributions toward the development of improved therapies for patients with affective disorders.
Furthermore, from my interactions with the counseling community and medical doctors, I know
that these professionals rely on tools and knowledge gained through basic research to treat their
patients. I, therefore, am confident that through research and active outreach, I can make a
significant and positive impact on enhancing human health.