During my undergraduate career at the University of Richmond, I was fortunate to be able to conduct independent research with Drs. Roni Kingsley and Valerie Kish. My focus was the expression of heat shock proteins in response to temperature stress in the octocoral, Leptogorgia virgulata.  Heat shock proteins are thought to be molecular chaperones that protect other proteins from degradation during times of stress. I performed western blots using antibodies for specific heat shock proteins and also conducted some electron microscopy.

I then moved to Chicago and worked for three years as a clinical research coordinator for the Chicago Center for Clinical Research, a division of Protocare Trials.  I primarily worked on trials for Alzheimer’s disease and psychiatric disorders, and acted as a liason between doctors, patients, and pharmaceutical companies.  My clinical trial experience left me with more questions than answers, and enticed me to explore the field of pharmacology.  I became more interested in the molecular mechanisms of drugs and decided to pursue graduate school.

Laboratory Rotations:

Regulation of Neuronal Nicotinic Receptors in IMR-32 Cells
Ken Kellar, Ph.D.

I studied the effect that nicotine and nerve growth factor (NGF) had on nicotinic acetylcholine receptors (nAChR) in the human neuroblastoma cell line, IMR-32. Using [3H]epibatidine in radioligand binding studies, I was able to quantify the amount of nAChR present in treated and untreated cells. I also learned the technique, 86Rubdium efflux, an assay that measures function. Labeled rubidium is pumped into the cell via the Na+/K+ ATPase and release through the nAChR channel is measured after stimulation with nicotinic agonists.

Amyloid-beta toxicity and nicotine protection in mouse primary cultured cortical neurons
Jarda Wroblewski, Ph.D.

I learned techniques of primary neuronal culture and then tested different parameters for amyloid-beta toxicity and protection with nicotine. Previous studies in Dr. Wroblewski’s lab showed that glial cells must be present for nicotine to exert a protective effect.  Since neurotrophic factors are not stored, they must be made immediately prior to release.  This allowed me to use microarray studies to look at mRNA levels in astrocytes after nicotine treatment to identify the factors that may be related to neuroprotection.

Effects of trauma on LRP
Bill Rebeck, Ph.D. (Department of Neuroscience)

In this rotation, I was interested in the lipoprotein related receptor (LRP) and effects of trauma on LRP. Through collaborations with other labs, I obtained rat brains that had been exposed to trauma: kainic acid seizures or diffuse traumatic brain injury (DTBI). I used slices of these brains to perform immunohistochemistry with antibodies for LRP. I was also interested in defining the regulation of LRP; thus I performed the luciferase assay to determine if the LRP promoter was affected by hypoxia.

Thesis Project:

Title
Ken Kellar, Ph.D.

For my doctoral thesis, I am studying the functional role of nAChRs in the rat visual system.  I plan to perform neurotransmitter release studies in the retina and areas of the brain where the optic nerve projects.  In addition, I hope to develop a human retinoblastoma cell line as a model of nAChR function and regulation.

 Publications:

• Kingsley RJ, Affif E, Cox BC, Kothari S, Kriechbaum K, Kuchinsky K, Neill AT, Puri AF, Kish VM. Expression of heat shock and cold shock proteins in the gorgonian Leptogorgia virgulata. Journal of Comparative Zoology 296(2): 98-107, 2003

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