Comparison of anti-apoptotic signalling by the insulin receptor and IGF-I receptor in preadipocytes and adipocytes.
We compared the effectiveness of insulin receptor (IR) and type I insulin-like growth factor (IGF) receptor (IGFR) cytoplasmic domains in mediating anti-apoptotic effects in 3T3-L1 preadipocytes and adipocytes. We used TrkC/IR and TrkC/IGFR chimeras, stably expressed in these cells and stimulated with neurotrophin-3 (NT-3), so as to avoid interference from endogenous receptors. After 24-h serum deprivation, 10% of preadipocytes and 2% of adipocytes appeared apoptotic as determined by fluorescence-activated cell sorter (FACS) analysis of cells stained with propidium iodide (PI) and Annexin V. When NT-3 was added, the two chimeras inhibited apoptosis to the same extent by 80% in preadipocytes and 50% in adipocytes. Mutation of juxtamembrane tyrosines (IR Y960F, IGFR Y950F) abrogated these anti-apoptotic effects. Qualitatively similar results were obtained by determination of viable rather than apoptotic cells. We conclude that IR and IGFR have equal potential to inhibit apoptosis in cell backgrounds, which are normally responsive to either IGF-I or insulin.
Ursø
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B
University of Cambridge, Department of Clinical Biochemistry, Addenbrooke's Hospital, Hills Road, CB2 2QR, Cambridge, UK.
Niesler
C U
CU
O'Rahilly
S
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Siddle
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K
D017667:Adipocytes / Q000378:metabolism*; D000818:Animals; D017304:Annexin A5 / Q000494:pharmacology; D017209:Apoptosis; D002460:Cell Line; D002470:Cell Survival; D003593:Cytoplasm / Q000378:metabolism; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D007328:Insulin / Q000378:metabolism; D055504:Insulin Receptor Substrate Proteins; D051379:Mice; D009154:Mutation; D009414:Nerve Growth Factors / Q000378:metabolism; D020933:Neurotrophin 3 / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011419:Propidium / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D017526:Receptor, IGF Type 1 / Q000378:metabolism*; D011972:Receptor, Insulin / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors
D017304:Annexin A5; D007328:Insulin; D055504:Insulin Receptor Substrate Proteins; C526295:Irs1 protein, mouse; D009414:Nerve Growth Factors; D020933:Neurotrophin 3; D010750:Phosphoproteins; D011419:Propidium; D017526:Receptor, IGF Type 1; D011972:Receptor, Insulin; D016650:Fluorescein-5-isothiocyanate; C073922:neurotrophin 4