Nitric oxide induction of neuronal endonuclease activity in programmed cell death - PubMed (original) (raw)

. 1999 Feb 1;246(2):290-300.

doi: 10.1006/excr.1998.4282.

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Nitric oxide induction of neuronal endonuclease activity in programmed cell death

A M Vincent et al. Exp Cell Res. 1999.

Abstract

Neuronal survival is intricately linked to the maintenance of intact DNA. In contrast, neuronal degeneration following nitric oxide (NO) exposure is dependent, in part, on the degradation of DNA through programmed cell death (PCD). We therefore investigated in primary rat hippocampal neurons the role of endogenous deoxyribonucleases, enzymes responsible for metabolically derived DNA cleavage, during NO-induced neurodegeneration. Twenty-four hours following exposure to the NO generators sodium nitroprusside (300 microM) and SIN-1 (300 microM), neuronal survival was reduced from approximately 88 to 23%. Treatment with aurintricarboxylic acid (1-100 microM), an endonuclease inhibitor, during NO exposure increased neuronal survival from 23 to 80% and decreased DNA fragmentation from 70 to 30% over a 24-h period. Enhancement of endonuclease activity alone with zinc chelation actively decreased neuronal survival from approximately 80% to approximately 34%. DNA digestion assays identified not only two constitutively active endonucleases, an acidic endonuclease (pH 4.0-7.0) and a calcium/magnesium-dependent endonuclease (pH 7.2-8.0), but also a NO-inducible magnesium-dependent endonuclease (pH 8.0). In the absence of endonuclease activity, DNA degradation did not occur during NO application, suggesting that endonuclease activity was a requisite pathway for NO-induced PCD. In addition, NO independently altered intracellular pH in ranges that were physiologically relevant for the activity of the endonucleases responsible for DNA degradation. Our identification and characterization of specific neuronal endonucleases suggest that the constitutive endonucleases may play a role in the initial stages of NO-induced PCD, but the subsequent "downstream" degradation of DNA may ultimately be dependent upon the NO-inducible endonuclease.

Copyright 1999 Academic Press.

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