It is our goal to continue our molecular and genetic analysis of the cha-1 gene complex from the soil nematode Caenorhabditis elegans. cha-1 is the structural gene for choline acetyltransferase (ChAT), the enzyme which synthesizes the neurotransmitter acetylcholine. Previously obtained evidence suggests that the ChAT protein in C. elegans may consist of multiple structural and functional domains, and we are interested in determining the organization of the gene. In addition, ChAT is considered a differentiation-specific marker for cholinergic neurons, and we wish to investigate its spatial and temporal control 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. We have recently used the transposable element Tcl to """"""""tag"""""""" and clone part of the cha-1 complex. A physical map of the gene will be constructed, and correlated with the genetic map already available. The complete nucleotide sequence of the gene will then be determined. Transcripts from the region will be studied by Northern analysis, S1 analysis, and the isolation of cDNA sequences. The transcriptional map thus constructed will be compared to the genetic map; this will test our published model of ChAT structure and function. The information obtained about wild-type gene structure and transcripts will then be used to analyze mutants defective in cholinergic function. Antibodies directed against ChAT will be produced, for use in biochemical and anatomical studies. 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 Alzheimer's disease, ALS, and depression.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM038679-07
Application #
3295283
Study Section
Neurology C Study Section (NEUC)
Project Start
1989-12-01
Project End
1994-08-31
Budget Start
1993-09-01
Budget End
1994-08-31
Support Year
7
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Oklahoma Medical Research Foundation
Department
Type
DUNS #
937727907
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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