Project: Research project

Project Details


Basic information concerning the mechanisms responsible for hypoxic cell
death is lacking. The overall goal of the proposed project is to define
the cellular mechanisms which are responsible for the onset of irreversible
injury and cell death using isolated rat hepatocytes as a model system.
Cell viability, cytosolic free calcium and pH, mitochondrial membrane
potential, lysosomal distribution and integrity, cytoskeletal status,
plasma membrane distribution and integrity, and cell surface morphology
will be evaluated in single individual hepatocytes during hypoxia by
quantitative digitized video microscopy (DVM). DVM will allow the direct
observation of the dynamics of organelles and molecules in living cells,
providing new information about the roles of these components in cellular
function. The onset of irreversible cell injury and cell death and the
progression or recovery from injury during reoxygenation will be determined
with respect to these cellular functions. Since increased cytosolic free
calcium, cell swelling, decreased energy supply, proteolysis and reactive
oxygen species have all been hypothesized to play a role in cell death
during hypoxia or following reoxygenation, pharmacologic stabilization
regimes utilizing calcium antagonists, anaerobic substrates, osmotic
agents, protease inhibitors, and scavengers of free oxygen radicals will be
assessed with regard to their ability to delay or prevent the onset of
irreversible injury and cell death. The role of the cytoskeleton in
hypoxic injury will be examined using agents which modulate cytoskeletal
structure and function. Additionally, cytoskeletal structure during
hypoxia will be visualized using immunocytochemistry in combination with
light and electron microscopy. Parallel experiments with suspensions of
hepatocytes will also be carried out using conventional biochemical
techniques. This project will provide fundamental, new information
regarding mechanisms responsible for the onset of hypoxic cell death. This
information will be important for the development of treatment modalities
effective in the preservation of cells and tissue during hypoxic episodes.
Effective start/end date5/1/874/30/16


  • Medicine(all)