Insulin-like growth factor-1-mediated protection from neuronal apoptosis is linked to phosphorylation of the pro-apoptotic protein BAD but not to inhibition of cytochrome c translocation in rat cerebellar neurons.
Cerebellar granule neurons cultured in the presence of serum and depolarizing potassium concentrations undergo apoptosis when switched to serum-free medium containing physiological potassium concentrations but remain viable after serum deprivation alone. Here, we show that potassium deprivation is associated with the dephosphorylation of the BCL-2-related BAD protein. Exposure to insulin-like growth factor-1 (IGF-1) inhibits both apoptosis and dephosphorylation of BAD. Both effects of IGF-1 do not depend on protein synthesis but are nullified by the phosphatidylinositol-3 kinase inhibitors, wortmannin and LY294002. In contrast to the treatment with cycloheximde, IGF-1 does not block the translocation of cytochrome c from mitochondria to the cytosol. Further, dephosphorylation of BAD alone does not appear to be sufficient to trigger apoptosis, since inhibition of protein synthesis by cycloheximide prevents apoptosis, but not BAD dephosphorylation, after potassium deprivation. These results suggest the coexistence of two parallel pathways, protein synthesis-dependent cytochrome c translocation and protein synthesis-independent dephosphorylation of BAD, both of which have to be activated to induce neuronal apoptosis.
Gleichmann
M
M
Department of Neurology, University of Tübingen, Medical School, Hoppe-Seyler-Strasse 3, D-72076, Tübingen, Germany.
Weller
M
M
Schulz
J B
JB
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism*; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism*; D002867:Chromones / Q000494:pharmacology; D003574:Cytochrome c Group / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000502:physiology*; D009025:Morpholines / Q000494:pharmacology; D009474:Neurons / Q000502:physiology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation; D011188:Potassium / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D051381:Rats; D000077191:Wortmannin; D051025:bcl-Associated Death Protein
D000730:Androstadienes; C494821:Bad protein, rat; D002352:Carrier Proteins; D002867:Chromones; D003574:Cytochrome c Group; D004791:Enzyme Inhibitors; D009025:Morpholines; D000081082:Phosphoinositide-3 Kinase Inhibitors; D019253:Proto-Oncogene Proteins c-bcl-2; D051025:bcl-Associated Death Protein; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D007334:Insulin-Like Growth Factor I; D011188:Potassium; D000077191:Wortmannin