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Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
15:43

Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes

Published on: January 7, 2013

Partial ATP depletion induces Fas- and caspase-mediated apoptosis in MDCK cells

L R Feldenberg1, S Thevananther, M del Rio

  • 1Division of Pediatric Nephrology, Yale University School of Medicine, New Haven, Connecticut 06520, USA.

The American Journal of Physiology
|June 11, 1999
PubMed

Insights

Partial ATP depletion in kidney cells triggers apoptosis through Fas and caspase pathways. This programmed cell death involves DNA fragmentation and is preventable with caspase inhibitors.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Renal Physiology

Background:

  • In vitro hypoxia/ischemia induces apoptosis in renal epithelial cells.
  • Mechanisms underlying this programmed cell death remain largely unknown.

Purpose of the Study:

  • To investigate the mechanisms of apoptosis induction in kidney cells due to partial ATP depletion.
  • To elucidate the roles of Fas and caspase pathways in this process.

Main Methods:

  • Madin-Darby canine kidney (MDCK) cells were subjected to partial ATP depletion.
  • Apoptosis was assessed via DNA fragmentation, morphological changes, and phosphatidylserine externalization.
  • Expression of Fas, Fas ligand, and FADD was analyzed.
  • Caspase activity (caspase-8, -1, -3) and PARP cleavage were measured.
  • Inhibitors of caspases and serine proteases were used to evaluate pathway involvement.

Main Results:

  • Partial ATP depletion (10-65%) induced apoptosis in MDCK cells in a duration-dependent manner.
  • ATP-depleted cells showed increased expression of Fas, Fas ligand, and FADD.
  • Fas-dependent apoptosis was confirmed in non-depleted cells using Fas monoclonal antibodies.
  • Cleavage of PARP and increased caspase-8 activity indicated caspase activation.
  • Apoptosis was significantly reduced by caspase inhibitors (caspase-8, -1, -3) but not serine protease inhibitors.

Conclusions:

  • Partial ATP depletion is sufficient to induce apoptosis in renal epithelial cells.
  • Fas-mediated signaling and caspase activation play critical roles in ATP depletion-induced apoptosis.
  • Targeting caspase pathways may offer therapeutic strategies for renal ischemia/hypoxia.

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