Genetic and metabolic status of NGF-deprived sympathetic neurons saved by an inhibitor of ICE family proteases

J Cell Biol. 1996 Dec;135(5):1341-54. doi: 10.1083/jcb.135.5.1341.

Abstract

Sympathetic neurons undergo programmed cell death (PCD) when deprived of NGF. We used an inhibitor to examine the function of interleukin-1 beta-converting enzyme (ICE) family proteases during sympathetic neuronal death and to assess the metabolic and genetic status of neurons saved by such inhibition. Bocaspartyl(OMe)-fluoromethylketone (BAF), a cell-permeable inhibitor of the ICE family of cysteine proteases, inhibited ICE and CPP32 (IC50 approximately 4 microM) in vitro and blocked Fas-mediated apoptosis in thymocytes (EC50 approximately 10 microM). At similar concentrations, BAF also blocked the NGF deprivation-induced death of rat sympathetic neurons in culture. Compared to NGF-maintained neurons, BAF-saved neurons had markedly smaller somas and maintained only basal levels of protein synthesis; readdition of NGF restored growth and metabolism. Although BAF blocked apoptosis in sympathetic neurons, it did not prevent the fall in protein synthesis or the increase in the expression of c-jun, c-fos, and other mRNAs that occur during neuronal PCD, implying that the ICE-family proteases function downstream of these events during PCD.NGF and BAF rescued sympathetic neurons with an identical time course, suggesting that NGF, in addition to inhibiting metabolic and genetic events associated with neuronal PCD, can act posttranslationally to abort apoptosis at a time point indistinguishable from the activation of cysteine proteases. Both poly-(ADP ribose) polymerase and pro-ICE and Ced-3 homolog-1 (ICH-1) appear to be cleaved in a BAF-inhibitable manner, although the majority of pro-CPP32 appears unchanged, suggesting that ICH-1 is activated during neuronal PCD. Potential implications of these findings for anti-apoptotic therapies are discussed.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Chloromethyl Ketones / pharmacology*
  • Animals
  • Apoptosis* / drug effects
  • Caspase 1
  • Caspase 2
  • Caspase 3
  • Caspases*
  • Cell Nucleus / metabolism
  • Cysteine Endopeptidases / metabolism*
  • Cysteine Proteinase Inhibitors / pharmacology*
  • DNA Fragmentation / drug effects
  • Gene Expression / drug effects
  • Genes, fos
  • Genes, jun
  • Mice
  • Mice, Inbred C57BL
  • Nerve Growth Factors / pharmacology
  • Nerve Growth Factors / physiology*
  • Neurons / cytology
  • Neurons / physiology*
  • Poly(ADP-ribose) Polymerases / metabolism
  • Proto-Oncogene Proteins c-fos / metabolism
  • Rats
  • Superior Cervical Ganglion / cytology
  • T-Lymphocytes / cytology
  • T-Lymphocytes / drug effects
  • Time Factors
  • fas Receptor / physiology

Substances

  • Amino Acid Chloromethyl Ketones
  • Cysteine Proteinase Inhibitors
  • Nerve Growth Factors
  • Proto-Oncogene Proteins c-fos
  • fas Receptor
  • Poly(ADP-ribose) Polymerases
  • Casp3 protein, mouse
  • Casp3 protein, rat
  • Caspase 2
  • Caspase 3
  • Caspases
  • Cysteine Endopeptidases
  • Caspase 1