? The purpose of this proposal is to continue molecular and genetic analysis of acetylcholine synthesis and packaging in the soil nematode Caenorhabditis elegans. These studies focus on three genes and proteins: cha-1 is the structural gene for choline acetyltransferase (CHAT), the enzyme which synthesizes the neurotransmitter acetylcholine; unc-17 is the gene which encodes the synaptic vesicle acetylcholine transporter (VAChT); and cho-1 encodes the plasma membrane choline high-affinity transporter (CHT). Two of these genes, cha-1 and unc-17, are tightly linked and define the """"""""cholinergic gene locus"""""""". C. elegans is being used for these studies because of its simple nervous system, its ease of genetic and molecular analysis, and the availability of DNA-mediated transformation techniques.
The specific aims i nclude development of a standardized sensitive assay system for quantifying cholinergic-mediated behavior, analysis of multiple cha-1 transcripts and multiple ChAT isoforms to determine their roles in ChAT function, isolation of additional cho-1 alleles and extragenic suppressors of cho-1, determining the role of phosphorylation in localization and function of the three cholinergic proteins, and (in collaboration with two other laboratories) structure-function studies of nematode and mammalian ChAT and VAChT. The results of these studies will elucidate the mechanisms used within nerve terminals to coordinate neurotransmitter supply and demand. Although this is basic research, it is clearly relevant to health, since alterations in acetylcholine metabolism and/or cholinergic function have been identified in many neurological and psychiatric disorders, including congenital myasthenic syndromes, Alzheimer's disease, and depression. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM038679-17
Application #
6732163
Study Section
Special Emphasis Panel (ZRG1-MDCN-1 (01))
Program Officer
Tompkins, Laurie
Project Start
1989-12-01
Project End
2007-03-31
Budget Start
2004-04-01
Budget End
2005-03-31
Support Year
17
Fiscal Year
2004
Total Cost
$331,100
Indirect Cost
Name
Oklahoma Medical Research Foundation
Department
Type
DUNS #
077333797
City
Oklahoma City
State
OK
Country
United States
Zip Code
73104
Mathews, Eleanor A; Mullen, Gregory P; Manjarrez, Jacob R et al. (2015) Unusual regulation of splicing of the cholinergic locus in Caenorhabditis elegans. Genetics 199:729-37
Mathews, Eleanor A; Mullen, Gregory P; Hodgkin, Jonathan et al. (2012) Genetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans. Genetics 192:1315-25
Duerr, Janet S; Han, He-Ping; Fields, Stephen D et al. (2008) Identification of major classes of cholinergic neurons in the nematode Caenorhabditis elegans. J Comp Neurol 506:398-408
Mullen, Gregory P; Mathews, Eleanor A; Vu, Mai H et al. (2007) Choline transport and de novo choline synthesis support acetylcholine biosynthesis in Caenorhabditis elegans cholinergic neurons. Genetics 177:195-204
Rand, James B (2007) Acetylcholine. WormBook :1-21
Sandoval, Gisela M; Duerr, Janet S; Hodgkin, Jonathan et al. (2006) A genetic interaction between the vesicular acetylcholine transporter VAChT/UNC-17 and synaptobrevin/SNB-1 in C. elegans. Nat Neurosci 9:599-601
Duerr, J S; Gaskin, J; Rand, J B (2001) Identified neurons in C. elegans coexpress vesicular transporters for acetylcholine and monoamines. Am J Physiol Cell Physiol 280:C1616-22
Lickteig, K M; Duerr, J S; Frisby, D L et al. (2001) Regulation of neurotransmitter vesicles by the homeodomain protein UNC-4 and its transcriptional corepressor UNC-37/groucho in Caenorhabditis elegans cholinergic motor neurons. J Neurosci 21:2001-14
Zhu, H; Duerr, J S; Varoqui, H et al. (2001) Analysis of point mutants in the Caenorhabditis elegans vesicular acetylcholine transporter reveals domains involved in substrate translocation. J Biol Chem 276:41580-7
Rand, J B; Duerr, J S; Frisby, D L (2000) Neurogenetics of vesicular transporters in C. elegans. FASEB J 14:2414-22

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