An interdisciplinary approach, involving molecular biology and electrophysiology, for cloning and studying structure and expression of genes coding for ion channels and neurotransmitter receptors expressed in mammalian brain and skeletal and cardiac muscle is proposed. Major emphasis will be placed on developing a new general method for cloning such genes for those cases in which the conventional approach through protein purification is not readily applicable. The method is based on electrophysiological assays for channels or receptors expressed on the oocyte surface by microinjection of mRNA into Xenopus oocytes, combined with cDNA cloning, and hybrid selection or hybrid arrest procedures. The first goals will be the isolation of genes coding for voltage sensitive calcium channels and for serotonin receptors. The chromosomal genes for the voltage sensitive sodium channels of brain, heart, and muscle will be isolated by standard methods from a presently available partial cDNA clone for the brain gene. Special emphasis will be placed on determining the molecular basis of the diversity of gene products for all the channels of interest, and to the significance of this diversity in signal processing. In all cases, high precision characterization of the electrophysiological properties of the channels expressed in abundance in the oocyte will be carried out by single channel patch clamping methods.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM010991-31
Application #
3268204
Study Section
Neurology C Study Section (NEUC)
Project Start
1977-06-01
Project End
1992-05-31
Budget Start
1990-06-01
Budget End
1992-05-31
Support Year
31
Fiscal Year
1990
Total Cost
Indirect Cost
Name
California Institute of Technology
Department
Type
Schools of Engineering
DUNS #
078731668
City
Pasadena
State
CA
Country
United States
Zip Code
91125
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