MEMBRANE PATHOLOGY IN RENAL CELL INJURY

Project: Research project

Project Details

Description

The objectives of the proposed research are to arrive at a better
understanding of how oxidative and anaerobic energy metabolism
in renal epithelial cells relate to the maintenance of cell
membrane integrity, and how impairment of energy metabolism
and the generation of ATP can cause cell damage by producing
pathological alterations in the lipid composition of cell
membranes. Experiments will be conducted on an established
renal epithelial cell line with proximal tubule characteristics,
primary cultured proximal tubule cells and freshly isolated
proximal tubule segments. Cellular energy metabolism will be
manipulated by the use of anoxia, or inhibitors of mitochondrial
oxidative phosphorylation, and by graded deletion of glucose
from the experimental medium. These manipulations will result
in graded levels of cell ATP, derived either from oxidative
metabolism or anaerobic glycolysis. The effects of these
manipulations on cells will be studied by measurement of
phospholipids, neutral lipids and cellular electrolytes, by
morphological examination and by a variety of techniques to
determine viability. After this characterization, studies will
focus on the use of supplemental or alternative glycolytic
substrates to sustain cell viability during injury, and the study of
the effect of cellular work on the evolution of cell injury. A
variety of biochemical and morphological techniques will be used
to achieve these objectives. Cells with impaired energy
metabolism will be metabolically characterized for oxygen
consumption, cytochrome a (a3) reduction, mitochondrial
membrane potential and glucose utilization. Alterations of lipid
metabolism will be studied by lipid extraction, chromatographic
separation and quantification of lipid mass. In addition, lipid
metabolism will be studied by radiolabelling techniques. Cell
electrolytes will be measured by atomic absorption spectroscopy
and electron probe microanalysis. Cell morphology will be
studied by light and electron microscopy. Cell viability will be
studies by determining the ability of cells to recover their ATP
levels, perform complex integrated functions such as protein
synthesis, and by electron microscopy of the cells following
restoration to a normal environment to allow recovery. These
studies may help devise effective strategies for the preservation
of cell integrity during injury caused by conditions characterized
by impaired energy metabolism, such as ischemia or anoxia.
StatusFinished
Effective start/end date7/1/876/30/10

Funding

  • National Institutes of Health: $481,271.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $350,120.00
  • National Institutes of Health: $464,562.00
  • National Institutes of Health: $481,470.00
  • National Institutes of Health
  • National Institutes of Health: $478,499.00
  • National Institutes of Health: $208,975.00
  • National Institutes of Health: $495,915.00
  • National Institutes of Health: $463,784.00
  • National Institutes of Health
  • National Institutes of Health: $517,446.00
  • 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: $250,724.00
  • National Institutes of Health
  • National Institutes of Health: $460,687.00

ASJC

  • Medicine(all)

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