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Robert F. Newkirk,
Professor (1980)
Ph.D. Colorado State
University/1972
MS Vrginia State University/1968
BS Livingstone College/1963Dept of Biological Sciences
Tennessee State University
3500 John Merritt Blvd
Nashville, TN 37209-1561
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The objectives of this research are to gain a better understanding of the
mechanisms supporting the function of the cholinergic synapse. Experiments
are being carried out to determine if the high affinity choline
cotransporter is found in two distinct populations in the cholinergic
terminals in the horseshoe crab. It is anticipated that one population
exists in the terminal membrane and a second exists in cytoplasmic vesicles
in the terminal. Efforts are being made to identify separate and
characterize two populations of choline cotransporters. Successful
completion of this project will have significant implications for studying
the molecular mechanisms undergirding membrane tracking on the choline
cotransporter. The following specific aims are being investigated:
1. To demonstrate that hemicholinium -2 (HC-3) binding is associated
selectively with cholinergic
terminals and that HC-3 binding is localized in the presynaptic membrane
of ChAT identified
terminals:
2. Top demonstrate that elevated K+ stimulated increase in
choline uptake is accomplished by a
quantifiable increase in HC-3 binding in cholinergic terminals:
3. To demonstrate that ACh vesicles and choline cotransporter vesicles are
different populations;
to isolate separable population of synaptic and synaptic-like vesicles -
one enriched with the
VAChT and ACh; the other enriched with the HC-3 binding sites.
4. To demonstrate physiological charateristics of the Ch cotransporter in
the isolated synaptic-like
vesicle population (fraction) enriched with HC-3 binding
5. To demonstrate PKC localization in cholinergic neurons and terminal. To
determine if a
specific isoform or multiple isoforms of PKC are localized in
cholinergic terminals. |
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