Evidence for impaired coupling of receptors to Gi protein in adipocytes from streptozocin-induced diabetic rats.
Adenosine and prostaglandins of the E series inhibit lipolysis in adipocytes by binding to cell surface receptors. This inhibition is mediated via Gi. It has been reported that Gi is almost absent in livers from diabetic rats. Therefore, we have evaluated the sensitivity of adipocytes from diabetic rats to the adenosine analogue N6-phenylisopropyl adenosine (PIA) and to prostaglandin E1 (PGE1). Diabetes was induced with streptozocin (65 mg/kg i.v.), and after 7 days, adipocytes were isolated. Lipolysis (measured in the presence of adenosine deaminase) was inhibited by PIA and PGE1 in both control and diabetic cells. However, the dose-response curves were markedly shifted to the right in the cells from diabetic rats. The IC50 for PIA was 0.30 +/- 0.02 nM in controls and 0.83 +/- 0.08 in diabetic rats (P less than 0.001), and the IC50 for PGE1 was 3.16 +/- 0.18 nM in controls and 5.26 +/- 0.57 nM in diabetic rats (P less than 0.02). These findings indicate decreased sensitivity to both adenosine and PGE1. Adipocyte membranes were isolated from control and diabetic rats. Adenosine receptors (measured by binding of 125I-labeled hydroxy-PIA) were not altered in cells from diabetic rats. However, the ability of Gpp(NH)p (a nonhydrolyzable GTP analogue) to inhibit adenosine-receptor binding was markedly decreased in membranes from diabetic rats, suggesting a change at the level of Gi. The alpha-subunits of Gi1, Gi2, Gi3, and Gs were quantitated on Western blots with a series of recently characterized anti-peptide antisera. This revealed that the amounts of each of these G proteins were normal in membranes from the diabetic rats.(ABSTRACT TRUNCATED AT 250 WORDS)
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Department of Internal Medicine, University of Texas Medical Branch, Galveston 77550.
Johnson
J L
JL
D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism*; D000527:Alprostadil / Q000494:pharmacology*; D000818:Animals; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000302:isolation & purification / Q000502:physiology*; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D007700:Kinetics; D008066:Lipolysis / Q000187:drug effects*; D008297:Male; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D010660:Phenylisopropyladenosine / Q000494:pharmacology*; D011458:Prostaglandins E / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D011982:Receptors, Prostaglandin / Q000187:drug effects / Q000502:physiology*; D018078:Receptors, Prostaglandin E; D011983:Receptors, Purinergic / Q000187:drug effects / Q000502:physiology*; D012016:Reference Values; D010566:Virulence Factors, Bordetella / Q000378:metabolism
D011458:Prostaglandins E; D011982:Receptors, Prostaglandin; D018078:Receptors, Prostaglandin E; D011983:Receptors, Purinergic; D010566:Virulence Factors, Bordetella; D009243:NAD; D010660:Phenylisopropyladenosine; D006165:Guanylyl Imidodiphosphate; D019204:GTP-Binding Proteins; D000527:Alprostadil