Date of Award

Fall 1997

Document Type

Thesis

Degree Name

Master of Science in Biomedical Engineering - (M.S.)

Department

Biomedical Engineering Committee

First Advisor

Peter Engler

Second Advisor

Vanessa H. Routh

Third Advisor

Joseph J. McArdle

Abstract

Of particular interest in this study were the electrical phenomena involving currents from the ATP sensitive K channel in isolated hypothalamic ventromedial nucleus (VMN) neurons. These neurons may play an important role in the glucose-sensing system of the body. Rat VMN neurons were isolated. For neuronal isolation, the proteolytic enzyme pronase was used. However, pronase was too harsh. The use of papain, another proteolytic enzyme, led to better neuronal morphology. The perforated patch clamp technique was used to gain access to the cell's interior. For membrane pore formation, nystatin was used; but seal formation was difficult. Amphotericin was used instead, and this milder antibiotic worked better at seal formation. Lastly, Na/K currents obtained from isolated neurons were amplified, recorded, and analyzed.

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