The long-term goal of the research described in this application is the elucidation of how chemical and electrical excitation of the plasma membrane affect gene expression in the neural control of acetylcholine sensitivity. Analysis of acetylcholine receptor regulation is important for a better understanding of neurological and neuromuscular disorders such as Alzheimer's disease or myasthenia gravis. The denervation-induced increase in acetylcholine receptor synthesis rate will be investigated in chick skeletal muscle. It is now known that during differentiation of myogenic cells and upon denervation of mature muscle fibers the transcription of the genes coding for the receptor subunits is activated. Regulatory elements in these genes that mediate stage- and tissue-specific expression will be identified by transfection of constructs containing sequences of interest as well as suitable reporter genes. The response of transfected cells to drugs that stimulate or block membrane activity will be measured and will permit identification of regulatory elements have already been identified in the upstream flanking regions of the alpha- and delta-subunit genes; future work will focus on other noncoding regions of these genes and on the gamma-subunit. Identification, isolation, and analysis of beta- and alpha-subunit genes will also be undertaken. The initial deletion mutation analysis of cis elements will be followed by more precise delineation using point mutations and footprinting techniques. Eventually, the developmental as well as stimulus-induced coordinated regulation of all subunits will be investigated in vitro and in transgenic animals.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS020233-05A1
Application #
3400496
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1985-07-01
Project End
1997-03-31
Budget Start
1990-04-01
Budget End
1991-03-31
Support Year
5
Fiscal Year
1990
Total Cost
Indirect Cost
Name
State University New York Stony Brook
Department
Type
Schools of Arts and Sciences
DUNS #
804878247
City
Stony Brook
State
NY
Country
United States
Zip Code
11794
Spinner, Daryl S; Liu, Shaohua; Wang, Shao-Wen et al. (2002) Interaction of the myogenic determination factor myogenin with E12 and a DNA target: mechanism and kinetics. J Mol Biol 317:431-45
Liu, S; Spinner, D S; Schmidt, M M et al. (2000) Interaction of MyoD family proteins with enhancers of acetylcholine receptor subunit genes in vivo. J Biol Chem 275:41364-8
Neville, C M; Choe, Y H; Lee, Y S et al. (1998) The E protein CTF4 and acetylcholine receptor expression in development and denervation supersensitivity. J Biol Chem 273:14046-52
Malik, S; Huang, C F; Schmidt, J (1995) The role of the CANNTG promoter element (E box) and the myocyte-enhancer-binding-factor-2 (MEF-2) site in the transcriptional regulation of the chick myogenin gene. Eur J Biochem 230:88-96
Huang, C F; Flucher, B E; Schmidt, M M et al. (1994) Depolarization-transcription signals in skeletal muscle use calcium flux through L channels, but bypass the sarcoplasmic reticulum. Neuron 13:167-77
Huang, C F; Lee, Y S; Schmidt, M M et al. (1994) Rapid inhibition of myogenin-driven acetylcholine receptor subunit gene transcription. EMBO J 13:634-40
Huang, C F; Neville, C M; Schmidt, J (1993) Control of myogenic factor genes by the membrane depolarization/protein kinase C cascade in chick skeletal muscle. FEBS Lett 319:21-5
Jia, H T; Tsay, H J; Schmidt, J (1992) Analysis of binding and activating functions of the chick muscle acetylcholine receptor gamma-subunit upstream sequence. Cell Mol Neurobiol 12:241-58
Neville, C M; Schmidt, J (1992) Expression of myogenic factors in skeletal muscle and electric organ of Torpedo californica. FEBS Lett 305:23-6
Neville, C M; Schmidt, M; Schmidt, J (1992) Response of myogenic determination factors to cessation and resumption of electrical activity in skeletal muscle: a possible role for myogenin in denervation supersensitivity. Cell Mol Neurobiol 12:511-27

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