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Streptozotocin-induced diabetes impairs G-protein linked signal transduction in vascular smooth muscle

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Abstract

The present studies were undertaken to examine if the impaired vascular function observed in diabetes is attributed to the altered levels of G-protein. Diabetes was induced in Sprague Dawley rats by a single intraperitoneal injection of streptozotocin (STZ) (60 mg/kg body wt) and after a period of 5 days, the aorta were used for adenylyl cyclase activity determination and protein quantification. A temporal relationship between the expression of Giα proteins and development of diabetes was also examined on day 1, 2, 3, 4 and 5 of injection of STZ. Blood glucose levels were significantly increased from day 1 in STZ-rats as compared to their counterpart control rats and reached to about 20 mM on 3rd day and 30 mM on 5th day. The expression of Giα-2 and Giα-3 proteins as determined by immunoblotting techniques was decreased by about 70 and 50% respectively in aorta from STZ rats compared to the control rats after 5 days of treatment, whereas 40% decrease in Giα-2 and Giα-3 was observed after 3rd day of STZ injection. On the other hand, the expression of Gsα was unaltered in STZ rats. In addition, the stimulatory effect of cholera toxin (CT) on GTP-mediated stimulation of adenylyl cyclase was not different in STZ as compared to the control group. However, the stimulatory effects of isoproterenol, glucagon, NaF and FSK on adenylyl cyclase activity were significantly enhanced in STZ rats as compared to control rats, whereas basal adenylyl cyclase activity was significantly lower in STZ-rats as compared to control rats. In addition, GTPγS inhibited FSK-stimulated adenylyl cyclase activity in concentration-dependent manner (receptor-independent functions of Giα) in control rats which was completely attenuated in STZ-rats. In addition, receptor-mediated inhibitions of adenylyl cyclase by angiotensin II, oxotremorine, atrial natriuretic peptide (ANP99–126) and C-ANP4–23 were also attenuated (receptor-dependent functions of Giα) in STZ-rats. These results indicate that aorta from diabetic rats exhibit decreased levels of cAMP and decreased expression of Giα. The decreased expression of Giα may be responsible for the altered responsiveness of adenylyl cyclase to hormonal stimulation and inhibition in STZ-rats. It may thus be suggested that the impaired adenylyl cyclase-Giα protein signaling may be one of the possible mechanisms responsible for the impaired vascular functions in diabetes.

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References

  1. Neer EJ: Heterotrimeric G proteins. Organizers of transmembrane signals. Cell 80: 249-257, 1995

    Google Scholar 

  2. Fleming JW, Wisler PL, Watanbe AM: Signal transduction by G-proteins in cardiac tissues. Circulation 85: 420-433, 1992

    Google Scholar 

  3. Murakami T, Yasuda H: Rat heart cell membranes contain three substrate for cholera toxin-catalyzed ADP-ribosylation and a single substrate for Pertussis toxin-catalyzed ADP-ribosylation. Biochem Biophys Res Commun 138: 1355-1361, 1986

    Google Scholar 

  4. Robishaw JD, Smigel MD, Gilman AG: Molecular basis for two forms of the G-protein that stimulate adenylate cyclase. J Biol Chem 261: 9587-9590, 1986

    Google Scholar 

  5. Bray P, Caster A, Simons C, Guo V, Puckett C, Hamholz J, Spiegel A, Nirenberg H: Human cDNA clones for four species of Gsá signal transduction protein. Proc Natl Acad Sci USA 83: 8893-8897, 1986

    Google Scholar 

  6. Itoh H, Kozaka, T, Magata S, Nakamura S, Katada T, Ui M, Iwai S, Ohtsuka E, Kawasaki H, Suzuki K, Kaziro Y: Molecular cloning and sequence determination of cDNAs for α subunits of the guanine nucleotide-binding proteins Gs, Gi and Go from brain. Proc Natl Acad Sci USA 83: 3776-3780, 1986

    Google Scholar 

  7. Itoh H, Toyama R, Kozasa T, Tsukamoto T, Matsuoka M, Kaziro Y: Presence of three distinct molecular species of Gi protein: A subunit structure of rat cDNA and human genomic DNAs. J Biol Chem 263: 6656-6664, 1988

    Google Scholar 

  8. Wong YH, Conklin BB, Bourne HR: G2-mediated hormonal inhibition of cyclic AMP accumulation. Science 255: 339-342, 1992

    Google Scholar 

  9. Yatani A, Mattera R, Codina J: The G-protein gated atrial K+ channels is stimulated by three distinct Giα subunits. Nature 336: 680-682, 1988

    Google Scholar 

  10. Simon MI, Strathmann MP, Gautam N, Science 252: 802-808, 1991

    Google Scholar 

  11. Cali JJ, Balcueva EA, Rybalkin I, Robishaw JD: Selective tissue distribution of G protein γ subunits, including a new form of the γ subunits identified by cDNA cloning. J Biol Chem 267: 24023-24027, 1992

    Google Scholar 

  12. Tang WJ, Gilman AG: Type-specific regulation of adenylyl cyclase by G protein βγ subunits. Science 254: 1500-1503 1991

    Google Scholar 

  13. Wickman KD, Iniguez-Liuhi JA, Davenport PA, Taussig R, Krapivinski GB, Linder ME, Gilman AG, Clapham DE: Recombinant G-protein βγ subunit activate the muscarinic-gated atrial potassium channel. Nature 368: 255-257, 1994

    Google Scholar 

  14. Tang WJ, Gilman AG: Adenylyl cyclase. Cell 70: 869-872, 1992

    Google Scholar 

  15. Premont RT, Chen J, Ma HW, Ponnapalli M, Iyengar R: Two members of a widely expressed subfamily of hormone-stimulated adenylyl cyclases. Proc Natl Acad Sci USA 89: 9809-9813, 1992

    Google Scholar 

  16. Katsushika S, Chen L, Kawabe JI, Nilakantan R, Halnon NJ, Homcy CJ, Ishikawa Y: Cloning and characterization of a sixth adenylyl cyclase isoform: type V and VI constitute a subgroup within the mammalian adenylyl cyclase family. Proc Natl Acad Sci USA 89: 8774-8778, 1992

    Google Scholar 

  17. Katz MA, Tada M, Kirchberger MA: Control of calcium transport in the myocardium by the cyclic AMP-protein kinase system. Adv Cyclic Nucleotide Res 5: 453-472, 1975

    Google Scholar 

  18. Keely SL Jr, Lincoln TM, Corbin JD: Interaction of acetylcholine and epinephrine on heart cyclic AMP-dependent protein kinase. Am J Physiol 234: H432-H438, 1978

    Google Scholar 

  19. Triner L, Vulliemoz Y, Verosky M, Habif DV, Nahas VV: Adenylyl cyclase phosphodiesterase system in arterial smooth muscle. Life Sci 11: 817-824, 1972

    Google Scholar 

  20. Bär HP: Cyclic nucleotides and smooth muscle. Adv Cyclic nucleotide Res 4: 195-237, 1974

    Google Scholar 

  21. Feldman AM, Cates AE, Veazey WB, Hershberger RE, Bristow MR, Baughman KL, Baumgartner WA, Dop CV: Increase of the 40,000 mol wt pertussis toxin substrate (G-protein) in the failing human heart. J Clin Invest 82: 189-197, 1988

    Google Scholar 

  22. Malbon CC, Rapiejko PJ, Watkins DC: Permissive hormone regulation of hormone-sensitive effector systems. Trends Pharmacol Sci 9: 33-36, 1988

    Google Scholar 

  23. Anand-Srivastava MB: Enhanced expression of inhibitory guanine nucleotide regulatory protein in spontaneously hypertensive rats: Relationship to adenylate cyclase inhibition. Biochem J 288: 79-85, 1992

    Google Scholar 

  24. Thibault C, Anand-Srivastava MB: Altered expression of G-protein mRNA in spontaneously hypertensive rats. FEBS Lett 313: 160-164, 1992

    Google Scholar 

  25. Anand-Srivastava MB, de Champlain J, Thibault C: DOCA-salt hypertensive rat hearts exhibit altered expression of G-proteins. Am J Hypertens 6: 72-75, 1993

    Google Scholar 

  26. Moxham CM, Malbon CC: Insulin action impaired by deficiency of the G-protein subunit Gi alpha 2. Nature 379: 840-844, 1996

    Google Scholar 

  27. Zheng XL, Guo J, Wang HY, Malbone CC: Expression of constitutively activated Gialpha 2 in vivo ameliorates streptozotocin induced diabetes. J Biol Chem 273: 23649-23651, 1998

    Google Scholar 

  28. Lynch CJ, Blackmore PF, Johnson EH, Wang RL, Krone PK, Exton JH: Guanine nucleotide binding regulatory proteins and adenylate cyclase in livers of streptozotocin-and BB/Wor-diabetic rats. Immuno-detection of Gs and Gi with antisera prepared against synthetic peptides. J Clin Invest 83: 2050-2062, 1989

    Google Scholar 

  29. Strassheim D, Palmer T, Houslay MD: Genetically acquired diabetes: Adipocyte guanine nucleotide regulatory protein expression and adenylyl cyclase regulation. Biochim Biophys Acta 1096: 121-126, 1991

    Google Scholar 

  30. Allgayer H, Bachman W, Hepp KD: Increased dose response relationship of plasma membrane adenylate cyclase to glucagon stimulation in diabetic rats. A possible role of guanylyl nucleotide-binding regulatory proteins. Diabetelogia 22: 464-467, 1982

    Google Scholar 

  31. Gawler D, Milligan G, Spiegel AM, Unson CG, Houslay MD: Abolition of expression of inhibitory guanine nucleotide regulatory protein Gi activity in diabetes. Nature 327: 229-232, 1987

    Google Scholar 

  32. Pilkis SJ, Exton JH, Johnson RA, Park CR: Effects of glucagon on cyclic AMP and carbohydrate metabolism in livers from diabetic rats. Biochim Biophys Acta 20: 250-267, 1974

    Google Scholar 

  33. Garber AJ: The impact of streptozoticin-induced diabetes mellitus on cyclic nucleotide regulation of skeletal muscle amino acid metabolism in the rat. J Clin Invest 65: 478-487, 1980

    Google Scholar 

  34. Palmer GC, Wilson GL, Chronister RB: Streptozotocin-induced diabetes produces alterations in adenylate cyclase in rat cerebrum, cerebral microvessels and retina. Life Sci 24: 365-374, 1983

    Google Scholar 

  35. Marcil J, de Champlain J, Anand-Srivastava MB: Overexpression of Gi-proteins precedes the development of DOCA-salt-induced hypertension: Relationship with adenylyl cyclase. Cardiovasc Res 39: 492-505, 1998

    Google Scholar 

  36. Salomon Y, Londos C, Rodbell MA: Highly sensitive assay. Annal Biochem 58: 541-548, 1974

    Google Scholar 

  37. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with folin-phenol reagent. J Biol Chem 193: 265-275, 1951

    Google Scholar 

  38. Jones DT, Reed RR: Molecular cloning of five GTP-binding protein cDNA species from rat olfactory neuroepithelium. J Biol Chem 262: 14241-14249, 1987

    Google Scholar 

  39. Anand-Srivastava MB, Srivastava KA, Cantin M: Pertussis toxin attenuates atrial natriuretic factor-mediated inhibitin of adneylyl cyclase. Involvement of inhibitory guanine nucleotide regulatory protein. J Biol Chem 262: 4931-4934, 1987

    Google Scholar 

  40. Anand-Srivastava MB, Sairam MR, Cantin M: Ring-deleted analogs of atrial factor inhibits adenylyl cyclase/cAMP system: Possible coupling of clearance atrial natriuretic factor receptors to adenylate cyclase/cAMP signal transduction system. J Biol Chem 265: 8566-8572, 1990

    Google Scholar 

  41. Anand-Srivastava MB: Angiotensin II receptors negatively coupled to adenylate cyclase in rat myocardial sarcolemma: Involvement of inhibitory guanine nucleotide protein. Biochem Pharmacol 38: 489-496, 1989

    Google Scholar 

  42. Bushfield M, Griffiths SL, Murphy GJ, Pyne NJ, Knowler JT, Milligen G, Parker PJ, Mollner S, Houslay MD: Diabetes-induced alterations in the expression, functioning and phosphorylation state of the inhibitory guanine nucleotide regulatory protein Giα-2 in hepatocytes. Biochem J 271: 365-372, 1990

    Google Scholar 

  43. Caro JF, Raju MS, Caro M, Lynch CJ, Poulos J, Exton JH, Thakkar JK: Evidence for altered 'cross-talk' between the insulin receptor and Gi proteins. J Cell Biochem 54: 309-319, 1994

    Google Scholar 

  44. Livingstone C, McLellan AR, McGregor M-A, Wilson A, Connell JMC, Small M, Milligan G, Paterson KR, Houslay MD: Altered G-protein expression and adenylate cyclase activity in platelets of non-insulin-dependent (NIDDM) male subjects. Biochim Biochem Acta 1096: 127-133, 1991

    Google Scholar 

  45. Hadjiconstantinou M, Ou ZX, Moroi-Fettersse, Neff NH: Apparent loss of Gi protein activity in diabetic retina. Eur J Pharmacol 149: 193-194, 1988

    Google Scholar 

  46. Weber LP, Macleod KM: Influence of streptozotocin diabetes on the alpha-1 adrenoceptor and associated G-protein in rat arteries. J Pharmacol Exp Ther 283: 1469-1478, 1997

    Google Scholar 

  47. Hattori Y, Matsuda N, Sato A, Watanuki S, Tomioka H, Kawasaki H, Kanno M: Predominant contribution of the G protein mediated mechanism to NaF-induced vascular contractions in diabetic rats: Association with an increased level of G (q alpha) expression. J Pharmacol Exp Ther 292: 761-768

  48. Swithers SB, Stewart RE, McCarty R: Binding sites for atrial natriuretic factor (ANF) in kidneys and adrenal glands of spontaneously hypertensive (SHR) rats. Life Sci 1673-1681, 1987

  49. Schiffrin EL: Decreased density of binding sites for atrial natriuretic peptide on platelets of patients with severe congestive heart failure. Clin Sci 74: 213-218, 1988

    Google Scholar 

  50. Strassheim D, Palmer T, Houslay MD: Diabetes abolishes the GTP-dependent, but not the nucleotide regulatory protein expression and adenylate cyclase regulation. Biochim Biophys Acta 1096: 121-126, 1991

    Google Scholar 

  51. Kowluru A, Kowluru RA, Yamaraki A: Functional alterations of G-proteins in diabetic rat retina: A possible explanation for the early visual abnormalities in diabetes mellitus. Diabetologia 35: 624-631, 1992

    Google Scholar 

  52. Limas C, Limas CJ: Reduced number of á-adrenergic receptors in the myocardium of spontaneously hypertensive rats. Biochem Biophys Res Commun 83: 710-714, 1978

    Google Scholar 

  53. Anand-Srivastava MB: Amiloride interacts with guanine nucleotide regulatory proteins and attenuates the hormonal inhibitin of adenylate cyclase. J Biol Chem 264: 9491-9496, 1989

    Google Scholar 

  54. Anand-Srivastava MB: Rat platelets from spontaneously hypertensive rats exhibit decreased expression of inhibitory guanine nucleotide regulatory protein: Relationship with adenylate cyclase activity. Circ Res 73: 1032-1039, 1993

    Google Scholar 

  55. Marcil J, Schiffrin EL, Anand-Srivastava MB: Aberrant adenylyl cyclase/cAMP signal transduction system and G protein levels in platelets from hypertensive patients: Improve with antihypertensive drug therapy. Hypertension 28: 83-90, 1996

    Google Scholar 

  56. Anand-Srivastava MB: Downregulation of atrial natriuretic peptide ANP-C receptor is associated with alterations in G-protein expression in A10 smooth muscle cells. Biochemistry 39: 6503-6513, 2000

    Google Scholar 

  57. Hildebrandt JD, Hanoune J, Birnbaumer J: Guanine nucleotide inhibition of cyc-S49 mouse lymphoma cell membrane adenylyl cyclase. J Biol Chem 257: 14273-14275, 1982

    Google Scholar 

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  1. Department of Physiology and Groupe de recherche sur le Système Nerveux Autonome, Faculty of Medicine, University of Montreal, Montreal, Quebec, Canada, H3C 3J7

    Shehla Hashim, Yi Yong Liu, Rui Wang & Madhu B. Anand-Srivastava

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  1. Shehla Hashim
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Hashim, S., Liu, Y.Y., Wang, R. et al. Streptozotocin-induced diabetes impairs G-protein linked signal transduction in vascular smooth muscle. Mol Cell Biochem 240, 57–65 (2002). https://doi.org/10.1023/A:1020652526803

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