This National Science Foundation Young Investigator project is in the general area of analytical and surface chemistry and in the subfields of bioanalysis and neurochemistry. During the tenure of this five-year award, Professor Sweedler and his students will develop the analytical instrumentation and methodology capable of identifying and quantifying neuronal releasates from a single nerve terminal and the contents of individual varicosities, and then use those techniques to better understand the mechanisms of cellular communication. Specific thrusts include the development of capillary zone electrophoresis/laser induced fluorescence instrumentation and methodology to assay and quantitate the contents of individual varicosities and vesicles; demonstration of the capabilities of that instrumentation using cultured neurons; study of the distribution of neuropeptides within individual neurons of the giant marine mollusk Aplysia; quantitative measurement of the peptide content of individual varicosities along a nerve process; measurement of the release of neuropeptides from a single nerve terminal; and determination of whether facilitation in Aplysia neurons is due to enhancement of neurotransmitter release. %%% The successful attainment of the goals of this project will afford protocols for the reliable quantitation of analytes in the zeptomole range, thereby extending the sensitivity of conventional cell sampling techniques by over a thousand fold. This research will also serve to further bridge the scientific domains of analytical chemistry and cellular neurobiology, lead to a description of the subcellular dynamics of neuronal signalling, and contribute to our basic understanding of the nervous system.
