Topic of the rotation:
Rationale for study and hypotheses tested:
For the
Training Grant progress report, for each rotation, please include the
following :
1)Summary description of the specific techniques that you learned and used
in the rotation
2) Summary of data generated
3) Any problems that you ran into and how you learned to deal with
them/troubleshoot/improve a method or protocol
4) Conclusions (did your data support the hypothesis, raise new questions,
point out new problems, etc)
Please write these in abstract form (see examples below).
Would you consider this rotation to be successful or not? Why:
EXAMPLES -
This format is required for the training grant reports each year
1) Summer
2004; Robert Yasuda, Ph.D., Assistant Professor, Pharmacology
To gain
experience in working in a molecular lab, and to investigate an area of
particular curiosity for me - NMDARs - I spent my summer working with Dr.
Robert Yasuda, investigating the interactions between NMDA receptors and
Eph/Ephrin receptors. Working closely with Dr. Yasuda, I learned lab
techniques including cell transfection, protein assays, immunoprecipitations,
Western blots, and using e-coli to 'grow' DNA. I was then able to work more
independently on these tasks. To investigate physical interactions between
NMDARs and Eph receptors, we transfected HEK293T and HEK293 cells with DNA
for different combinations of Eph receptors and NR1 and NR2B subunits of
NMDARs. Through immunoprecipitations and Western blots, we found
interactions between Eph receptors and NMDARS, even in the presence of
NR2B. In HEK cells, likely due to the relatively great expression of the
transfected genes, it was not necessary to stimulate Eph receptors with
Ephrins to see this interaction. In addition to molecular techniques, this
research experience required critical reading of primary literature on Eph/ephrins
and NMDAR, expanding my understanding of both. At the end of this rotation,
I presented my findings at Neurolunch, a regularly meeting series at
Georgetown.
2) Fall
2004; Guinevere Eden, Ph.D., Associate Professor, Pediatrics; Director of
the Center for the Study of Learning
In
addition to analyzing behavioral data for an ongoing project in the lab
looking at the effectiveness of different interventions for developmental
dyslexia, I spent my fall rotation extending a meta-analysis method
developed by Peter Turkeltaub (a former IPN student) to the study of
pseudowords. This project included a thorough literature search to find
relevant studies, an increased understanding of imaging techniques,
selection of inclusion and exclusion criteria, and interpretation of the
findings. I am currently working on writing up this project for
publication. First, we performed a meta-analysis of pseudoword reading
compared to baseline. This allows not only an overview of the most robust,
common findings across studies, it allows insight into aloud and silent
pseudoword reading differences. In addition to this first meta-analysis, we
performed a meta-analysis of pseudoword reading greater than real word
reading. This meta-analysis found two areas of common activation, with
particularly intriguing results suggesting the importance of baseline word
type (regular vs irregular words). Finally, we used this meta-analysis
technique to investigate the effects of task type. This lab itself uses
implicit reading tasks, so we were particularly interested in the overlap
these studies show with more traditional explicit reading tasks. Our
results have led to a pilot study to investigate this question at an
individual subject level, thus spanning the wide range of analysis levels
possible.
2) Fall
2004; Josh Corbin, Ph.D., Assistant Professor, Neuroscience
The focus of this
rotation was the study of genetic contributions to the development of the
amygdaloid nuclear complex. We hypothesized that boundaries of gene
expression delineating specific nuclei within the amygdaloid complex would
signal the alteration or absence of these nuclei in Nkx2.1-/-
mice. These mice completely lack a proliferating cell population of the
ventral telencephalon known as the medial ganglionic eminence, whose progeny
migrate throughout the developing brain and primarily differentiate into
inhibitory interneurons. During the rotation I was involved in breeding
mice, dissecting embryos at embryonic days 12.5, 15.5, and 18.5, embedding
tissue and sectioning on a cryostat, genotyping with PCR, and probing gene
expression via in situ hybridization. We examined expression of
Pax6, a gene that marks cells that are derived from the dorsal telencephalon
and which also migrate to the amygdala. In a single comparison, the pattern
of Pax6 expression in the Nkx2.1-/- was the same as the control
in the basomedial nucleus of the amygdala, and somewhat expanded in the
vicinity of the basolateral amygdala. This expansion may be linked to the
absence of ventrally-derived cells or their gene-products. Further
characterization of gene-expression in these knockouts is underway in the
Corbin lab.
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