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. 1992 Feb 15;282(Pt 1):99–106. doi: 10.1042/bj2820099

Cellular mechanism of the insulin-like effect of growth hormone in adipocytes. Rapid translocation of the HepG2-type and adipocyte/muscle glucose transporters.

J W Tanner 1, K A Leingang 1, M M Mueckler 1, K C Glenn 1
PMCID: PMC1130895  PMID: 1371670

Abstract

The cellular mechanism whereby growth hormone (GH) acutely stimulates adipocyte glucose uptake was studied in cultures of primary rat adipocytes differentiated in vitro. Preadipocytes were isolated by collagenase digestion of inguinal fat-pads from young rats and were differentiated in the presence of 3-isobutyl-1-methylxanthine, insulin and dexamethasone. The development of an adipocyte morphology (i.e. lipid inclusions) was observed over 6 days after initiation of differentiation. Coincident with this phenotypic change was an increase in glyceraldehyde-3-phosphate dehydrogenase (GPDH) activity and in cellular content of the HepG2-type (Glut1) and adipocyte/muscle (Glut4) glucose transporter isoforms as determined by Western immunoblotting of total cellular protein. Age-matched undifferentiated cells expressed the Glut1 transporter and low levels of GPDH, but neither accumulated lipid nor exhibited measurable expression of the Glut4 protein. On day 6 after the initiation of differentiation, GH and insulin stimulated 2-deoxy[14C]glucose uptake in a dose- and time-dependent fashion in adipocytes cultured under serum-free conditions for at least 15 h. Western-blot analysis of subcellular fractions revealed that both GH and insulin rapidly (within 20 min) stimulated translocation of the Glut1 and Glut4 proteins from a low-density microsomal fraction to the plasma membrane. Confirmatory evidence was provided in immunocytochemical experiments utilizing antisera directed against the C-terminal region of the Glut4 protein and a fluorescein isothiocyanate-labelled second antibody. Observation of the cells via confocal laser microscopic imaging was consistent with glucose transporter redistribution from an intracellular region to the plasma membrane after treatment with GH or insulin. On the basis of these data, we suggest that the insulin-like effect of GH on adipocyte glucose transport involves translocation of the Glut1 and Glut4 proteins to the plasma membrane. Furthermore, stimulation of glucose-transporter translocation by both GH and insulin may indicate a common cell signalling element between the adipocyte GH and insulin receptors or, alternatively, the existence of multiple cellular mechanisms for stimulating glucose-transporter translocation.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baumbach W. R., Horner D. L., Logan J. S. The growth hormone-binding protein in rat serum is an alternatively spliced form of the rat growth hormone receptor. Genes Dev. 1989 Aug;3(8):1199–1205. doi: 10.1101/gad.3.8.1199. [DOI] [PubMed] [Google Scholar]
  2. Benovic J. L., Bouvier M., Caron M. G., Lefkowitz R. J. Regulation of adenylyl cyclase-coupled beta-adrenergic receptors. Annu Rev Cell Biol. 1988;4:405–428. doi: 10.1146/annurev.cb.04.110188.002201. [DOI] [PubMed] [Google Scholar]
  3. Björntorp P., Karlsson M., Pettersson P., Sypniewska G. Differentiation and function of rat adipocyte precursor cells in primary culture. J Lipid Res. 1980 Aug;21(6):714–723. [PubMed] [Google Scholar]
  4. Calderhead D. M., Kitagawa K., Tanner L. I., Holman G. D., Lienhard G. E. Insulin regulation of the two glucose transporters in 3T3-L1 adipocytes. J Biol Chem. 1990 Aug 15;265(23):13801–13808. [PubMed] [Google Scholar]
  5. Campbell R. M., Kostyo J. L., Scanes C. G. Lipolytic and antilipolytic effects of human growth hormone, its 20-kilodalton variant, a reduced and carboxymethylated derivative, and human placental lactogen on chicken adipose tissue in vitro. Proc Soc Exp Biol Med. 1990 Apr;193(4):269–273. doi: 10.3181/00379727-193-43034. [DOI] [PubMed] [Google Scholar]
  6. Carter-Su C., Rozsa F. W., Wang X., Stubbart J. R. Rapid and transitory stimulation of 3-O-methylglucose transport by growth hormone. Am J Physiol. 1988 Nov;255(5 Pt 1):E723–E729. doi: 10.1152/ajpendo.1988.255.5.E723. [DOI] [PubMed] [Google Scholar]
  7. Carter-Su C., Stubbart J. R., Wang X. Y., Stred S. E., Argetsinger L. S., Shafer J. A. Phosphorylation of highly purified growth hormone receptors by a growth hormone receptor-associated tyrosine kinase. J Biol Chem. 1989 Nov 5;264(31):18654–18661. [PubMed] [Google Scholar]
  8. Cushman S. W., Wardzala L. J. Potential mechanism of insulin action on glucose transport in the isolated rat adipose cell. Apparent translocation of intracellular transport systems to the plasma membrane. J Biol Chem. 1980 May 25;255(10):4758–4762. [PubMed] [Google Scholar]
  9. Czech M. P. The nature and regulation of the insulin receptor: structure and function. Annu Rev Physiol. 1985;47:357–381. doi: 10.1146/annurev.ph.47.030185.002041. [DOI] [PubMed] [Google Scholar]
  10. Daughaday W. H., Rotwein P. Insulin-like growth factors I and II. Peptide, messenger ribonucleic acid and gene structures, serum, and tissue concentrations. Endocr Rev. 1989 Feb;10(1):68–91. doi: 10.1210/edrv-10-1-68. [DOI] [PubMed] [Google Scholar]
  11. Davidson M. B. Effect of growth hormone on carbohydrate and lipid metabolism. Endocr Rev. 1987 May;8(2):115–131. doi: 10.1210/edrv-8-2-115. [DOI] [PubMed] [Google Scholar]
  12. Douen A. G., Jones M. N. The role of insulin receptor sulphydryl groups in insulin binding and cellular response in rat adipocytes. J Recept Res. 1990;10(1-2):45–59. doi: 10.3109/10799899009064657. [DOI] [PubMed] [Google Scholar]
  13. Eriksson J., Gause-Nilsson I., Lönnroth P., Smith U. The insulin-like effect of growth hormone on insulin-like growth factor II receptors is opposed by cyclic AMP. Evidence for a common post-receptor pathway for growth hormone and insulin action. Biochem J. 1990 Jun 1;268(2):353–357. doi: 10.1042/bj2680353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Foster C. M., Shafer J. A., Rozsa F. W., Wang X. Y., Lewis S. D., Renken D. A., Natale J. E., Schwartz J., Carter-Su C. Growth hormone promoted tyrosyl phosphorylation of growth hormone receptors in murine 3T3-F442A fibroblasts and adipocytes. Biochemistry. 1988 Jan 12;27(1):326–334. doi: 10.1021/bi00401a049. [DOI] [PubMed] [Google Scholar]
  15. Fukumoto H., Kayano T., Buse J. B., Edwards Y., Pilch P. F., Bell G. I., Seino S. Cloning and characterization of the major insulin-responsive glucose transporter expressed in human skeletal muscle and other insulin-responsive tissues. J Biol Chem. 1989 May 15;264(14):7776–7779. [PubMed] [Google Scholar]
  16. Fukumoto H., Seino S., Imura H., Seino Y., Eddy R. L., Fukushima Y., Byers M. G., Shows T. B., Bell G. I. Sequence, tissue distribution, and chromosomal localization of mRNA encoding a human glucose transporter-like protein. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5434–5438. doi: 10.1073/pnas.85.15.5434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Glenn K. C., Rose K. S., Krivi G. G. Somatotropin antagonism of insulin-stimulated glucose utilization in 3T3-L1 adipocytes. J Cell Biochem. 1988 Aug;37(4):371–383. doi: 10.1002/jcb.240370405. [DOI] [PubMed] [Google Scholar]
  18. Goodman H. M. Multiple effects of growth hormone on lipolysis. Endocrinology. 1968 Aug;83(2):300–308. doi: 10.1210/endo-83-2-300. [DOI] [PubMed] [Google Scholar]
  19. Gould G. W., Derechin V., James D. E., Tordjman K., Ahern S., Gibbs E. M., Lienhard G. E., Mueckler M. Insulin-stimulated translocation of the HepG2/erythrocyte-type glucose transporter expressed in 3T3-L1 adipocytes. J Biol Chem. 1989 Feb 5;264(4):2180–2184. [PubMed] [Google Scholar]
  20. Green H., Kehinde O. Spontaneous heritable changes leading to increased adipose conversion in 3T3 cells. Cell. 1976 Jan;7(1):105–113. doi: 10.1016/0092-8674(76)90260-9. [DOI] [PubMed] [Google Scholar]
  21. Hauner H., Entenmann G., Wabitsch M., Gaillard D., Ailhaud G., Negrel R., Pfeiffer E. F. Promoting effect of glucocorticoids on the differentiation of human adipocyte precursor cells cultured in a chemically defined medium. J Clin Invest. 1989 Nov;84(5):1663–1670. doi: 10.1172/JCI114345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hauser S. D., McGrath M. F., Collier R. J., Krivi G. G. Cloning and in vivo expression of bovine growth hormone receptor mRNA. Mol Cell Endocrinol. 1990 Sep 10;72(3):187–200. doi: 10.1016/0303-7207(90)90143-v. [DOI] [PubMed] [Google Scholar]
  23. Isgaard J., Möller C., Isaksson O. G., Nilsson A., Mathews L. S., Norstedt G. Regulation of insulin-like growth factor messenger ribonucleic acid in rat growth plate by growth hormone. Endocrinology. 1988 Apr;122(4):1515–1520. doi: 10.1210/endo-122-4-1515. [DOI] [PubMed] [Google Scholar]
  24. James D. E., Brown R., Navarro J., Pilch P. F. Insulin-regulatable tissues express a unique insulin-sensitive glucose transport protein. Nature. 1988 May 12;333(6169):183–185. doi: 10.1038/333183a0. [DOI] [PubMed] [Google Scholar]
  25. James D. E., Strube M., Mueckler M. Molecular cloning and characterization of an insulin-regulatable glucose transporter. Nature. 1989 Mar 2;338(6210):83–87. doi: 10.1038/338083a0. [DOI] [PubMed] [Google Scholar]
  26. Janicot M., Lane M. D. Activation of glucose uptake by insulin and insulin-like growth factor I in Xenopus oocytes. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2642–2646. doi: 10.1073/pnas.86.8.2642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kayano T., Burant C. F., Fukumoto H., Gould G. W., Fan Y. S., Eddy R. L., Byers M. G., Shows T. B., Seino S., Bell G. I. Human facilitative glucose transporters. Isolation, functional characterization, and gene localization of cDNAs encoding an isoform (GLUT5) expressed in small intestine, kidney, muscle, and adipose tissue and an unusual glucose transporter pseudogene-like sequence (GLUT6). J Biol Chem. 1990 Aug 5;265(22):13276–13282. [PubMed] [Google Scholar]
  28. Kayano T., Fukumoto H., Eddy R. L., Fan Y. S., Byers M. G., Shows T. B., Bell G. I. Evidence for a family of human glucose transporter-like proteins. Sequence and gene localization of a protein expressed in fetal skeletal muscle and other tissues. J Biol Chem. 1988 Oct 25;263(30):15245–15248. [PubMed] [Google Scholar]
  29. Leung D. W., Spencer S. A., Cachianes G., Hammonds R. G., Collins C., Henzel W. J., Barnard R., Waters M. J., Wood W. I. Growth hormone receptor and serum binding protein: purification, cloning and expression. Nature. 1987 Dec 10;330(6148):537–543. doi: 10.1038/330537a0. [DOI] [PubMed] [Google Scholar]
  30. Li C. H., Evans H. M. THE ISOLATION OF PITUITARY GROWTH HORMONE. Science. 1944 Mar 3;99(2566):183–184. doi: 10.1126/science.99.2566.183. [DOI] [PubMed] [Google Scholar]
  31. Mathews L. S., Enberg B., Norstedt G. Regulation of rat growth hormone receptor gene expression. J Biol Chem. 1989 Jun 15;264(17):9905–9910. [PubMed] [Google Scholar]
  32. Mathews L. S., Norstedt G., Palmiter R. D. Regulation of insulin-like growth factor I gene expression by growth hormone. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9343–9347. doi: 10.1073/pnas.83.24.9343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Miller W. L., Martial J. A., Baxter J. D. Molecular cloning of DNA complementary to bovine growth hormone mRNA. J Biol Chem. 1980 Aug 25;255(16):7521–7524. [PubMed] [Google Scholar]
  34. Mueckler M., Caruso C., Baldwin S. A., Panico M., Blench I., Morris H. R., Allard W. J., Lienhard G. E., Lodish H. F. Sequence and structure of a human glucose transporter. Science. 1985 Sep 6;229(4717):941–945. doi: 10.1126/science.3839598. [DOI] [PubMed] [Google Scholar]
  35. Norstedt G., Möller C. Growth hormone induction of insulin-like growth factor I messenger RNA in primary cultures of rat liver cells. J Endocrinol. 1987 Oct;115(1):135–139. doi: 10.1677/joe.0.1150135. [DOI] [PubMed] [Google Scholar]
  36. Négrel R., Grimaldi P., Ailhaud G. Establishment of preadipocyte clonal line from epididymal fat pad of ob/ob mouse that responds to insulin and to lipolytic hormones. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6054–6058. doi: 10.1073/pnas.75.12.6054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Oka Y., Asano T., Shibasaki Y., Kasuga M., Kanazawa Y., Takaku F. Studies with antipeptide antibody suggest the presence of at least two types of glucose transporter in rat brain and adipocyte. J Biol Chem. 1988 Sep 15;263(26):13432–13439. [PubMed] [Google Scholar]
  38. RODBELL M. METABOLISM OF ISOLATED FAT CELLS. I. EFFECTS OF HORMONES ON GLUCOSE METABOLISM AND LIPOLYSIS. J Biol Chem. 1964 Feb;239:375–380. [PubMed] [Google Scholar]
  39. Ramsay T. G., White M. E., Wolverton C. K. Insulin-like growth factor 1 induction of differentiation of porcine preadipocytes. J Anim Sci. 1989 Sep;67(9):2452–2459. doi: 10.2527/jas1989.6792452x. [DOI] [PubMed] [Google Scholar]
  40. Schwartz Y., Goodman H. M., Yamaguchi H. Refractoriness to growth hormone is associated with increased intracellular calcium in rat adipocytes. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6790–6794. doi: 10.1073/pnas.88.15.6790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Silverman M. S., Mynarcik D. C., Corin R. E., Haspel H. C., Sonenberg M. Antagonism by growth hormone of insulin-sensitive hexose transport in 3T3-F442A adipocytes. Endocrinology. 1989 Nov;125(5):2600–2604. doi: 10.1210/endo-125-5-2600. [DOI] [PubMed] [Google Scholar]
  42. Simpson I. A., Cushman S. W. Hormonal regulation of mammalian glucose transport. Annu Rev Biochem. 1986;55:1059–1089. doi: 10.1146/annurev.bi.55.070186.005211. [DOI] [PubMed] [Google Scholar]
  43. Suzuki K., Kono T. Evidence that insulin causes translocation of glucose transport activity to the plasma membrane from an intracellular storage site. Proc Natl Acad Sci U S A. 1980 May;77(5):2542–2545. doi: 10.1073/pnas.77.5.2542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tordjman K. M., Leingang K. A., James D. E., Mueckler M. M. Differential regulation of two distinct glucose transporter species expressed in 3T3-L1 adipocytes: effect of chronic insulin and tolbutamide treatment. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7761–7765. doi: 10.1073/pnas.86.20.7761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tordjman K. M., Leingang K. A., Mueckler M. Differential regulation of the HepG2 and adipocyte/muscle glucose transporters in 3T3L1 adipocytes. Effect of chronic glucose deprivation. Biochem J. 1990 Oct 1;271(1):201–207. doi: 10.1042/bj2710201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Waters M. J., Barnard R. T., Lobie P. E., Lim L., Hamlin G., Spencer S. A., Hammonds R. G., Leung D. W., Wood W. I. Growth hormone receptors--their structure, location and role. Acta Paediatr Scand Suppl. 1990;366:60–72. doi: 10.1111/j.1651-2227.1990.tb11600.x. [DOI] [PubMed] [Google Scholar]
  47. Zorzano A., Wilkinson W., Kotliar N., Thoidis G., Wadzinkski B. E., Ruoho A. E., Pilch P. F. Insulin-regulated glucose uptake in rat adipocytes is mediated by two transporter isoforms present in at least two vesicle populations. J Biol Chem. 1989 Jul 25;264(21):12358–12363. [PubMed] [Google Scholar]

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