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J Clin Invest. 1989 Jan; 83(1): 127–136.
PMCID: PMC303652

Identification of persistent defects in insulin receptor structure and function capillary endothelial cells from diabetic rats.


Insulin actions and receptors were studied in capillary endothelial cells cultured from diabetic BB rats and their nondiabetic colony mates. The endothelial cells from diabetic rats of 2 mo duration had persistent biological and biochemical defects in culture. Compared with normal rats, endothelial cells from diabetic rats grew 44% more slowly. Binding studies of insulin and insulin-like growth factor I (IGF-I) showed that cells from diabetic rats had 50% decrease of insulin receptor binding (nondiabetic: 4.6 +/- 0.7; diabetic: 2.6 +/- 0.4% per milligram protein, P less than 0.01), which was caused by a 50% decrease in the number of binding sites per milligram protein, whereas IGF-I binding was not changed. Insulin stimulation of 2-deoxy-glucose uptake and alpha-aminoisobutyric acid uptake were also severely impaired with a 80-90% decrease in maximal stimulation, in parallel with a 62% decrease in insulin-stimulated autophosphorylation (P less than 0.05). 125I-insulin cross-linking revealed an 140-kD alpha subunit of the insulin receptor similar to that in cells from nondiabetic rats, although bands at greater than 200 kD were also detected. The molecular weight of the insulin receptor beta subunit (by SDS-PAGE) was smaller in cells from diabetic than from normal rats (88-90 vs. 95 kD). Neuraminadase treatment of the partially purified insulin receptors decreased the molecular weight of the insulin receptors from nondiabetic rats to a greater degree than its diabetic counterpart. In contrast, Northern blot analysis of insulin receptor mRNAs using human cDNA probes revealed two species of 9.4 and 7.2 kb with no difference in mRNA abundance between cells from diabetic and nondiabetic rats. We conclude that the exposure of capillary endothelial cells to a diabetic milieu in vivo can cause specific and persistent changes in the insulin receptor and insulin action.

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Selected References

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  • Kohner EM. The problems of retinal blood flow in diabetes. Diabetes. 1976;25(2 Suppl):839–844. [PubMed]
  • Mogensen CE. Glomerular filtration rate and renal plasma flow in short-term and long-term juvenile diabetes mellitus. Scand J Clin Lab Invest. 1971 Sep;28(1):91–100. [PubMed]
  • Cohen MP, Surma ML, Wu VY. In vivo biosynthesis and turnover of glomerular basement membrane in diabetic rats. Am J Physiol. 1982 Apr;242(4):F385–F389. [PubMed]
  • Vracko R, Benditt EP. Restricted replicative life-span of diabetic fibroblasts in vitro: its relation to microangiopathy. Fed Proc. 1975 Jan;34(1):68–70. [PubMed]
  • Goldstein S, Moerman EJ, Soeldner JS, Gleason RE, Barnett DM. Diabetes mellitus and genetic prediabetes. Decreased replicative capacity of cultured skin fibroblasts. J Clin Invest. 1979 Mar;63(3):358–370. [PMC free article] [PubMed]
  • Bowersox JC, Sorgente N. Altered growth kinetics of dermal fibroblasts and arterial smooth muscle cells from spontaneously diabetic BB rats. Diabetes. 1985 Jul;34(7):628–633. [PubMed]
  • Freidenberg GR, Henry RR, Klein HH, Reichart DR, Olefsky JM. Decreased kinase activity of insulin receptors from adipocytes of non-insulin-dependent diabetic subjects. J Clin Invest. 1987 Jan;79(1):240–250. [PMC free article] [PubMed]
  • Comi RJ, Grunberger G, Gorden P. Relationship of insulin binding and insulin-stimulated tyrosine kinase activity is altered in type II diabetes. J Clin Invest. 1987 Feb;79(2):453–462. [PMC free article] [PubMed]
  • Sinha MK, Pories WJ, Flickinger EG, Meelheim D, Caro JF. Insulin-receptor kinase activity of adipose tissue from morbidly obese humans with and without NIDDM. Diabetes. 1987 May;36(5):620–625. [PubMed]
  • Burant CF, Treutelaar MK, Buse MG. Diabetes-induced functional and structural changes in insulin receptors from rat skeletal muscle. J Clin Invest. 1986 Jan;77(1):260–270. [PMC free article] [PubMed]
  • King GL, Goodman AD, Buzney S, Moses A, Kahn CR. Receptors and growth-promoting effects of insulin and insulinlike growth factors on cells from bovine retinal capillaries and aorta. J Clin Invest. 1985 Mar;75(3):1028–1036. [PMC free article] [PubMed]
  • Like AA, Butler L, Williams RM, Appel MC, Weringer EJ, Rossini AA. Spontaneous autoimmune diabetes mellitus in the BB rat. Diabetes. 1982;31(Suppl 1 Pt 2):7–13. [PubMed]
  • Sima AA, Chakrabarti S, Garcia-Salinas R, Basu PK. The BB-rat--an authentic model of human diabetic retinopathy. Curr Eye Res. 1985 Oct;4(10):1087–1092. [PubMed]
  • Brown DM, Steffes MW, Thibert P, Azar S, Mauer SM. Glomerular manifestations of diabetes in the BB rat. Metabolism. 1983 Jul;32(7 Suppl 1):131–135. [PubMed]
  • Gimbrone MA., Jr Culture of vascular endothelium. Prog Hemost Thromb. 1976;3:1–28. [PubMed]
  • Hachiya HL, Carpentier JL, King GL. Comparative studies on insulin-like growth factor II and insulin processing by vascular endothelial cells. Diabetes. 1986 Oct;35(10):1065–1072. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Ishibashi F, Hidaka H, Howard BV. Glucose enhancement of insulin action: elevated glucose levels increase insulin stimulation of 2-deoxyglucose uptake in cultured human fibroblasts. J Clin Endocrinol Metab. 1982 Jan;54(1):34–39. [PubMed]
  • Martin MS, Pohl SL. Insulin-induced insulin resistance of alpha-aminoisobutyric acid transport in cultured human skin fibroblasts. J Biol Chem. 1979 Oct 25;254(20):9976–9978. [PubMed]
  • Rechler MM, Podskalny JM, Goldfine ID, Wells CA. DNA synthesis in human fibroblasts: stimulation by insulin and by nonsuppressible insulin-like activity (NSILA-S). J Clin Endocrinol Metab. 1974 Sep;39(3):512–521. [PubMed]
  • Kasuga M, White MF, Kahn CR. Phosphorylation of the insulin receptor in cultured hepatoma cells and a solubilized system. Methods Enzymol. 1985;109:609–621. [PubMed]
  • Han JH, Stratowa C, Rutter WJ. Isolation of full-length putative rat lysophospholipase cDNA using improved methods for mRNA isolation and cDNA cloning. Biochemistry. 1987 Mar 24;26(6):1617–1625. [PubMed]
  • Aviv H, Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. [PMC free article] [PubMed]
  • Ullrich A, Bell JR, Chen EY, Herrera R, Petruzzelli LM, Dull TJ, Gray A, Coussens L, Liao YC, Tsubokawa M, et al. Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. Nature. 313(6005):756–761. [PubMed]
  • Zapf J, Schoenle E, Froesch ER. Insulin-like growth factors I and II: some biological actions and receptor binding characteristics of two purified constituents of nonsuppressible insulin-like activity of human serum. Eur J Biochem. 1978 Jun 15;87(2):285–296. [PubMed]
  • Jialal I, Crettaz M, Hachiya HL, Kahn CR, Moses AC, Buzney SM, King GL. Characterization of the receptors for insulin and the insulin-like growth factors on micro- and macrovascular tissues. Endocrinology. 1985 Sep;117(3):1222–1229. [PubMed]
  • Goldstein BJ, Muller-Wieland D, Kahn CR. Variation in insulin receptor messenger ribonucleic acid expression in human and rodent tissues. Mol Endocrinol. 1987 Nov;1(11):759–766. [PubMed]
  • Lorenzi M, Montisano DF, Toledo S, Barrieux A. High glucose induces DNA damage in cultured human endothelial cells. J Clin Invest. 1986 Jan;77(1):322–325. [PMC free article] [PubMed]
  • Viberti G, Pickup JC, Bilous RW, Keen H, Mackintosh D. Correction of exercise-induced microalbuminuria in insulin-dependent diabetics after 3 weeks of subcutaneous insulin infusion. Diabetes. 1981 Oct;30(10):818–823. [PubMed]
  • Rosenbloom AL, Silverstein JH, Lezotte DC, Richardson K, McCallum M. Limited joint mobility in childhood diabetes mellitus indicates increased risk for microvascular disease. N Engl J Med. 1981 Jul 23;305(4):191–194. [PubMed]
  • Goldstein S, Littlefield JW, Soeldner JS. Diabetes mellitus and aging: diminished planting efficiency of cultured human fibroblasts. Proc Natl Acad Sci U S A. 1969 Sep;64(1):155–160. [PMC free article] [PubMed]
  • Howard BV, Fields RM, Mott DM, Savage PJ, Nagulesparan M, Bennett PH. Diabetes and cell growth--lack of differences in growth characteristics of fibroblasts from diabetic and nondiabetic Pima Indians. Diabetes. 1980 Feb;29(2):119–124. [PubMed]
  • Raizada MK, Tan G, Fellows RE. Fibroblastic cultures from the diabetic db/db mouse. Demonstration of decreased insulin receptors and impaired responses to insulin. J Biol Chem. 1980 Oct 10;255(19):9149–9155. [PubMed]
  • Berliner JA, Frank HJ, Karasic D, Capdeville M. Lipoprotein-induced insulin resistance in aortic endothelium. Diabetes. 1984 Nov;33(11):1039–1044. [PubMed]
  • King GL, Buzney SM, Kahn CR, Hetu N, Buchwald S, Macdonald SG, Rand LI. Differential responsiveness to insulin of endothelial and support cells from micro- and macrovessels. J Clin Invest. 1983 Apr;71(4):974–979. [PMC free article] [PubMed]
  • Corkey RF, Corkey BE, Gimbrone MA., Jr Hexose transport in normal and SV40-transformed human endothelial cells in culture. J Cell Physiol. 1981 Mar;106(3):425–434. [PubMed]
  • Kahn CR. Insulin resistance, insulin insensitivity, and insulin unresponsiveness: a necessary distinction. Metabolism. 1978 Dec;27(12 Suppl 2):1893–1902. [PubMed]
  • Kahn CR. The molecular mechanism of insulin action. Annu Rev Med. 1985;36:429–451. [PubMed]
  • Megyesi K, Kahn CR, Roth J, Neville DM, Jr, Nissley SP, Humbel RE, Froesch ER. The NSILA-s receptor in liver plasma membranes. Characterization and comparison with the insulin receptor. J Biol Chem. 1975 Dec 10;250(23):8990–8996. [PubMed]
  • King GL, Kahn CR, Rechler MM, Nissley SP. Direct demonstration of separate receptors for growth and metabolic activities of insulin and multiplication-stimulating activity (an insulinlike growth factor) using antibodies to the insulin receptor. J Clin Invest. 1980 Jul;66(1):130–140. [PMC free article] [PubMed]
  • Banskota NK, Carpentier JL, King GL. Processing and release of insulin and insulin-like growth factor I by macro- and microvascular endothelial cells. Endocrinology. 1986 Nov;119(5):1904–1913. [PubMed]
  • Okamoto M, White MF, Maron R, Kahn CR. Autophosphorylation and kinase activity of insulin receptor in diabetic rats. Am J Physiol. 1986 Nov;251(5 Pt 1):E542–E550. [PubMed]
  • King GL, Johnson SM. Receptor-mediated transport of insulin across endothelial cells. Science. 1985 Mar 29;227(4694):1583–1586. [PubMed]

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