Degradation of the type I inositol 1,4,5-trisphosphate receptor by caspase-3 in SH-SY5Y neuroblastoma cells undergoing apoptosis

J Neurochem. 2000 Nov;75(5):1852-61. doi: 10.1046/j.1471-4159.2000.0751852.x.

Abstract

The type I inositol 1,4,5-trisphosphate (IP(3)) receptor is selectively down-regulated in several neurodegenerative diseases, including Alzheimer's disease, Huntington's chorea, and ischemia, all conditions in which apoptotic neuronal loss occurs. In the present study, we used a neuronal cell line, human neuroblastoma SH-SY5Y cells, to investigate whether the levels of IP(3) receptor are changed during apoptosis in these cells. Following induction of apoptosis by staurosporine, the immunoreactivity of the type I IP(3) receptor in microsome preparations from SH-SY5Y cells was reduced within 2 h, with a further reduction during subsequent hours. Immunoblot analyses, using antibodies to poly(ADP-ribose) polymerase and spectrin breakdown products, revealed proteolysis of these caspase-3 substrates within 3 h, confirming that IP(3) receptor cleavage is an early consequence of apoptosis. In vitro incubation of SH-SY5Y microsomes or immunopurified IP(3) receptor from rat cerebellum with recombinant caspase-3 led to generation of immunoreactive breakdown products similar to those observed in intact cells, suggesting that the type I IP(3) receptor is a potential substrate for caspase-3. Preincubation of the neuroblastoma cells with the caspase-3 inhibitor Z-Asp-Glu-Val-Asp-fluoromethyl ketone prevented IP(3) receptor degradation. These results show that the type I IP(3) receptor is a substrate for caspase-3 in neuronal cells and indicate that apoptotic down-regulation of IP(3) receptor levels may contribute to the pathology of neurodegenerative conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs / genetics
  • Apoptosis*
  • Calcium Channels / genetics
  • Calcium Channels / metabolism*
  • Caspase 3
  • Caspases / metabolism*
  • Cysteine Proteinase Inhibitors / pharmacology
  • Down-Regulation / drug effects
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Inositol 1,4,5-Trisphosphate Receptors
  • Neuroblastoma / genetics
  • Neuroblastoma / metabolism*
  • Neuroblastoma / pathology
  • Oligopeptides / pharmacology
  • Poly(ADP-ribose) Polymerases / metabolism
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Staurosporine / antagonists & inhibitors
  • Staurosporine / pharmacology
  • Tumor Cells, Cultured

Substances

  • Calcium Channels
  • Cysteine Proteinase Inhibitors
  • Enzyme Inhibitors
  • ITPR1 protein, human
  • Inositol 1,4,5-Trisphosphate Receptors
  • Oligopeptides
  • Receptors, Cytoplasmic and Nuclear
  • benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone
  • Poly(ADP-ribose) Polymerases
  • CASP3 protein, human
  • Casp3 protein, rat
  • Caspase 3
  • Caspases
  • Staurosporine