Inhibition of p38 mitogen-activated protein kinase by insulin in cultured fetal neurons.
Insulin supports the survival and differentiation of many types of fetal neurons. To determine if mitogen-activated protein (MAP) kinases play a role in mediating the neurotrophic actions of insulin, we identified the MAP kinases present in fetal chick forebrain neurons and examined their regulation by insulin. Cell extracts were fractionated on Mono Q columns, and phosphotransferase activity was measured using myelin basic protein as the substrate. In control neurons, four peaks of MAP kinase activity were resolved. Peaks I, II, and IV were identified by immunoblotting as c-Jun N-terminal kinase (JNK), extracellular signal-related kinase (ERK), and p38 MAP kinase, respectively. Neurons treated with insulin showed a dramatic decrease, 80-90%, in p38 MAP kinase activity without significant changes in the other MAP kinase activities. Insulin decreased the phosphotyrosine content of p38 MAP kinase with maximal effects observed within 5 min. Pretreatment of neurons with sodium orthovanadate blocked the ability of insulin to inhibit the tyrosine phosphorylation and activity of p38 MAP kinase, suggesting that activation of a tyrosine or dual specific phosphatase is necessary for the inhibition of p38 MAP kinase by insulin. Since p38 MAP kinase has been recently implicated in neuronal cell apoptosis, negative regulation of this kinase by insulin may be critical for the neurotrophic actions of insulin.
Heidenreich
K A
KA
Department of Pharmacology, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA.
Kummer
J L
JL
D051698:Activating Transcription Factor 2; D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002645:Chickens; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology*; D048031:JNK Mitogen-Activated Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D009474:Neurons / Q000201:enzymology*; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D019000:Phosphotyrosine / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases
D051698:Activating Transcription Factor 2; D017362:Cyclic AMP Response Element-Binding Protein; D004791:Enzyme Inhibitors; D007328:Insulin; D014157:Transcription Factors; D019000:Phosphotyrosine; D014638:Vanadates; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D010749:Phosphoprotein Phosphatases