The goal of this research project is to continue molecular, genetic and biochemical analyses of the char-1 gene complex from the soil nematode Caenorhabditis elegans. Char-1 is the structural gene for choline acetyltransferase, the enzyme which synthesizes the neurotransmitter acetylcholine. Previously obtained evidence suggests that the choline acetyltransferase protein in C. elegans may consists of multiple structural and functional domains, and there is interest in determining the organization of the gene and the structure of the protein. In addition, investigations on the spatial and temporal control of choline acetyltransferase will be performed during neural development. C. elegans is being used for these studies because of its simple nervous system, its ease of genetic analysis, and the availability of the technique of "transposon tagging" to clone genes from this organism. This recent approach has been utilized to "tag" and clone part of the cha-1 complex. Antibodies directed against specific regions of the choline acetyltransferase molecule will be isolated. The isolated regions will be used in biochemical studies of choline acetyltransferase structure and in anatomical studies to analyze the development of acetylcholine-utilizing neurons. The information obtained about the normal gene and enzyme will be used to analyze mutants defective in neural structure and neurotransmitter function.
