BIOPHYSICS OF GAP JUNCTION CHANNELS

Project: Research project

Project Details

Description

Gap junctions are low resistance channels that in excitable tissues provide
a pathway for impulse propagation and synchronous activation; in
inexcitable cells their role is presumably in exchange of metabolites or
signalling molecules. Considerable preliminary information is available
(largely funded by this grant in prior years) on both short and longterm
regulation of gap junctions. We intend to combine techniques of molecular
biology, immunology and electrophysiology in order to study expression of
gap junction protein and gating of gap junctions between cells of several
mammalian tissues. Pairs of isolated adult and cultured neonatal cells as
well as various cell lines, will be used as experimental models. In
addition we shall use methods of recording from gap junction membranes
isolated from rat hepatocytes. An antibody has been obtained which blocks
coupling between cell pairs from several tissues and has demonstrated
reactivity against gap junction protein. Using monospecific polyclonal
antibodies, relative levels of junctional protein will be determined by
immuno-precipitation and immunoblot techniques and compared to junctional
conductance (gj) under a variety of conditions expected to affect
expression. We will also correlate covalent protein modifications
(including phosphorylation) on gj, junctional permeability, and their
sensitivity to gating stimuli. These experiments should finally begin to
answer questions of how gap junctions are regulated and what role they play
in propagation of electrical and metabolic activity in physiological and
pathological states.
StatusFinished
Effective start/end date1/1/901/1/90

Funding

  • National Institute of Neurological Disorders and Stroke

ASJC

  • Biophysics
  • Physiology
  • Biochemistry
  • Cell Biology
  • Histology

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