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Papers In Press, published online ahead of print April 30, 2002
Sperm Biology, National Institute of Immunology, New Delhi, Delhi 110067
Corresponding Author: cshaha{at}nii.res.in
Reactive oxygen species are important regulators of protozoal infection. Promastigotes of L. donovani, the causative agent of Kala-azar, undergo an apoptosis-like death upon exposure to H2O2. The present study shows that upon activation of death response by H2O2, a dose and time dependent loss of mitochondrial membrane potential occurs. This loss is accompanied by a depletion of cellular glutathione but cardiolipin content or thiol oxidation status remains unchanged. ATP levels are reduced within the first 60 min of exposure as a result of mitochondrial membrane potential loss. A tight link exists between changes in cytosolic Ca2+ homeostasis and collapse of the mitochondrial membrane potential, but the dissipation of the potential is independent of elevation of cytosolic Na+ and mitochondrial Ca2+. Partial inhibition of cytosolic Ca2+ increase achieved by chelating extracellular or intracellular Ca2+ by the use of appropriate agents result in significant rescue of the fall of the mitochondrial membrane potential and apoptosis-like death. It is further demonstrated that the increase in cytosolic Ca2+ is an additive result of release of Ca2+ from intracellular stores as well as by influx of extracellular Ca2+ through flufenamic acid sensitive non-selective cation channels, contribution of the latter being larger. Mitochondrial changes do not involve opening of the mitochondrial transition pore as cyclosporin A is unable to prevent mitochondrial membrane potential loss. Antioxidant like N-acetylcysteine is able to inhibit the fall of the mitochondrial membrane potential and prevent apoptosis-like death. Together, these findings show the importance of non-selective cation channels in regulating the response of L. donovani promastigotes to oxidative stress that trigger downstream signaling cascades leading to apoptosis-like death.
J. Biol. Chem, 10.1074/jbc.M201961200
Submitted on February 27, 2002
Revised on April 30, 2002
Accepted on April 30, 2002
Increase in cytosolic Ca2+ levels through the activation of non-selective cation channels induced by oxidative stress cause mitochondrial depolarization leading to apoptosis-like death in L. donovani promastigotes
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