REGULATION OF THE SODIUM CHANNEL IN EXCITABLE CELLS

Project: Research project

Description

The voltage-dependent sodium channel is the basis of electrical
excitability in mammalian nerve and muscle cells. This proposal decribes
experiments to elucidate the structure of the rat sodium channel, to
measure expression of the sodium channel gene in response to neuronal
growth factors, and to identify and characterize regulatory sequences in
the gene. The structural gene (cDNA) for the rat muscle sodium channel
will be isolated from a bacteriophage expression library by screening with
polyclonal and monoclonal antibodies directed against purified sodium
channel proteins. The nucleotide sequence of the muscle cDNA clones will
be used to deduce the primary structure of the sodium channel proteins.
Northern blot and S1 nuclease hybridization assays will be performed to
determine whether muscle and nerve sodium channels are identical. A
sensitive assay system for measuring changes in sodium channel mRNA
production will be established using an NGF-inducible cell line (PC-12) and
the characterized cDNA clones as probes. These probes will be used in
Northern blot assays to detect changes in sodium channel mRNA levels after
treatment of PC-12 cells with NGF. If an effect on transcription of the
sodium channel gene is observed, experiments will be performed to identify
the NGF-responsive sequences in the gene. These studies involve
constructing expression vectors containing the sodium channel gene fused to
bacterial gene sequences. The recombinants will be introduced into PC-12
cells and assayed for fusion gene products in the presence and absence of
NGF. Because cAMP may be involved in the mechanism by which NGF exerts its
effects on PC-12 cells, cAMP will also be tested for its ability to
regulate expression of the sodium channel gene. These experiments will
provide a framework for understanding the molecular mechanisms underlying
the development of excitability in neuronal cells.
StatusFinished
Effective start/end date9/1/854/30/15

Funding

  • National Institutes of Health: $295,780.00
  • National Institutes of Health: $220,567.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $308,220.00
  • National Institutes of Health: $317,467.00
  • National Institutes of Health: $279,000.00
  • National Institutes of Health: $330,138.00
  • National Institutes of Health
  • National Institutes of Health: $270,138.00
  • National Institutes of Health: $352,945.00
  • National Institutes of Health
  • National Institutes of Health: $304,652.00
  • National Institutes of Health: $330,138.00
  • National Institutes of Health
  • National Institutes of Health: $270,679.00
  • National Institutes of Health: $333,507.00
  • National Institutes of Health: $295,323.00
  • National Institutes of Health: $318,583.00
  • National Institutes of Health: $287,163.00
  • National Institutes of Health: $348,039.00
  • National Institutes of Health
  • National Institutes of Health: $60,864.00
  • National Institutes of Health: $336,875.00

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Sodium Channels
Genes
Neurogenesis
Gene Expression
DNA
Transgenic Mice
Phenotype
Nerve Growth Factor
MicroRNAs
Action Potentials
Nucleic Acid Regulatory Sequences
Chromatin
Nervous System
Stem Cells
Embryonic Stem Cells
Membranes
Cell Line
Complementary DNA
DNA-Binding Proteins
Pluripotent Stem Cells

ASJC

  • Medicine(all)
  • Neuroscience(all)