DIGITAL FLUORESCENCE IMAGING SYSTEM

Project: Research project

Project Details

Description

This facility is designed for the general purpose of correlating optical
and electrophysiological measurements. The optical measurements that this
equipment would permit include spatial measurements of intracellular pH and
pCa, determination of membrane potentials in multicellular preparations by
voltage sensitive dyes, quantitation of dye fluxes within and between cells
resolution of cell volume and secretion changes and localization of
antibody binding to subcellular compartments. Electrophysiological
measurements will include resting and action potentials, junctional
conductances and ion concentrations as measured by ion selective
electrodes. Project 1 will evaluate gating mechanism and membrane
trafficking of gap junctions in cardiac myocytes, astroglial cells and
hepatocytes isolated from adult mammalian tissues. Sensitivity of
junctional conductance to intracellular H and Ca will be determined.
Fluorescent antibodies will be used to monitor internalization and possible
reuse. Label applied intracellularly may determine whether formation is by
redistribution of surface molecules or insertion from the cytoplasm. Other
studies will concern drug effects on cardiac myocytes in respect to changes
in intracellular ions and the concomitant changes in contractility.
Contractility of hepatocyte canaliculi will also be evaluated. Project 2
will determine gap junction channel properties by quantitative comparison
of dye fluxes and electrical conductance in various cell types. Problems
addressed will include all-or-none channel closure and interactions of
permeant molecules with channel walls. Project 3 will evaluate factors in
death and growth of cultured sympathetic neurons. Changes in neuritic
voltages, pH, pCa and redox state will be measured during normal growth and
during injury caused by starvation and anoxia. Project 4 will study
intracellular pH and intracellular communication between cells in the renal
collecting ducts with particular attention to the mitochondria-rich cells
that are presumed to secrete acid. These cells are surrounded by principal
cells to which they may not be coupled. Comparison will be made to other
regions of the kidney where coupling is presumed to occur. Project 5 will
concern second messengers and membrane trafficking in cells and cell lines
derived from kidney. Question to be addressed include temporal and spatial
resolution of Ca generation and time course of response to antidiuretic
hormone.
StatusFinished
Effective start/end date1/1/906/13/90

Funding

  • National Center for Research Resources

ASJC

  • Nephrology

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