Summary
125I-Insulin binding and insulin stimulation of glycogen synthase were examined in fibroblasts cultured from nine Type 1 (insulin-dependent) diabetic patients with age of onset of <42 years. In all cases specific insulin binding was qualitatively and quantitatively normal. Total 125I-insulin binding was elevated in cells from three patients with early onset diabetes (two with onset before age 1 year) due to an increase in ‘non-specific’ binding. When the ability of insulin to stimulate the conversion of the glucose-6-phosphate dependent to the glucose-6-phosphate independent form of glycogen synthase was measured, all cell lines responded, albeit to differing degrees. In general, the response of cells from diabetic donors was more variable than that of control fibroblasts. A slightly lower level of cellular glycogen was evident in the cells of the diabetic patients, and this was mirrored in slightly higher levels of the independent form of the enzyme. The average maximal level of the independent form of the enzyme also was higher in the diabetic patients' cells. Fibroblasts from one of the patients with very early onset diabetes had glycogen synthase levels that were markedly lower than in any other cell line examined. In summary, fibroblasts cultured from Type 1 diabetic patients do not show major defects in either insulin binding or action. A suggestion of subtle differences in the cells from the diabetic patients, particularly those with very early onset, is evident, however. Whether these are secondary to some primary genetic defect or represent some selection during culture remains to be determined.
Article PDF
Similar content being viewed by others
References
Dörfler H, Wieczorek A, Wolfram G, Zöllner N (1977) Binding of insulin to fibroblasts in lipoatrophic diabetes. Res Exp Med 170: 161–168
D'Ercole AJ, Underwood L, Groelke J, Plet A (1979) Leprechaunism: Studies of the relationship among hyperinsulinism, insulin resistance, and growth retardation. J Clin Endocrinol Metab 48: 495–502
Kobayashi M, Olefsky JM, Elders J, Mako ME, Givens BD, Schwedie HK, Fisler RH, Hintz RL, Horner JA, Rubenstein AH (1978) Insulin resistance due to a defect distal to the insulin receptor: Demonstration in a patient with leprechaunism. Proc Natl Acad Sci USA 75: 3469–3473
Schilling E, Rechler M, Grunfeld C, Rosenberg A (1979) Primary defect of insulin receptors in skin fibroblasts cultured from an infant with leprechaunism and insulin resistance. Proc Natl Acad Sci USA 76: 5877–5881
Raizada M, Tan G, Fellows R (1980) Fibroblast cultures from the diabetic db/db mouse. J Biol Chem 255: 9149–9155
Podskalny JM, Kahn CR (1980) Insulin activates glycogen synthase in cultured human fibroblasts. Diabetes 29: 724–729
Lawrence JC, Guinovart J, Larner J (1977) Activation of rat adipocyte glycogen synthase by insulin. J Biol Chem 252: 444–450
Alpers J (1966) The influence of hexose and insulin on glycogen synthase in hela cells. J Biol Chem 241: 217–222
Huang K (1979) Regulation of glycogen synthase activity in choriocarcinoma cells. Eur J Biochem 95: 477–485
Freychet P, Kahn CR, Roth J, Neville Jr DM, (1972) Insulin interaction with liver plasma membranes. J Biol Chem 247: 3953–3961
Rechler M, Podskalny J, Goldfine I, Wells C (1974) DNA synthesis in human fibroblasts: stimulation by insulin and by non-suppressible insulin-like activity. J Clin Endocrinol Metab 39: 512–521
Rechler MM, Podskalny JM (1976) Insulin receptors in cultured human fibroblasts. Diabetes 25: 250–255
Thomas J, Schlender K, Larner J (1968) A rapid filter paper assay for UDP glucose-glycogen glucosyltransferase, including an improved biosynthesis of UDP-14C-glucose. Anal Biochem 25: 486–499
Huang K, Cabib E (1973) Yeast glycogen synthase in the glucose-6-phosphate-independent form: a case of cold liability without major changes in molecular size. Biochem Biophys Acta 302: 240–248
Lo S, Russell JC, Taylor AW (1970) Determination of glycogen in small tissue samples. J App Physiol 28: 234–236
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–269
Podskalny J, Kahn CR (1982) Cell culture studies on patients with extreme insulin resistance. I. Receptor defects on cultured fibroblasts. J Clin Endocrinol Metab 54: 261–268
Podskalny J, Kahn CR (1982) Cell culture studies on patients with extreme insulin resistance. II. Abnormal biological responses in cultured fibroblasts. J Clin Endocrinol Metab 54: 269–275
Goldstein S, Littlefield JW, Soeldner JS (1979) Diabetes mellitus and aging: diminished plating efficiency of cultured human fibroblasts. Proc Natl Acad Sci USA 64:155–160
Goldstein S, Moerman EJ, Soeldner JS, Gleason RE, Barnett DM (1979) Diabetes mellitus and genetic prediabetics: decreased replicative capactiy of cultured skin fibroblasts. J Clin Invest 63: 358–370
Vracko R, Benditt EP (1975) Restricted replicative life-span of diabetic fibroblasts in vitro: its relation to micorangiopathy. Fed Proc 34:68–70
Goldstein S, Moerman E, Soeldner JS, Gleason RE, Barnett DM (1978) Chronologic and physiologic age affect replicative lifespan of fibroblasts from diabetic, pre-diabetic, and normal donors. Science 199: 781–782
Gleason RE, Goldstein S (1978) Age affect and replicative lifespan of fibroblasts of diabetic, prediabetic, and normal donors: another look at the data. Science 212:1217–1218
Rowe DW, Starman BJ, Fujimoto WY, Williams RH (1977) Abnormalities in proliferation and protein synthesis in skin fibroblast cultures from patients with diabetes mellitus. Diabetes 26: 284–290
Cooper JT, Goldstein S (1974) De novo synthesis of lipids and incorporation of oleic acid into cultured human fibroblasts from diabetics and normal controls. Atherosclerosis 20: 41–50
Kohn RR, Hensse S (1977) Abnormal collagen in cultures of fibroblasts from human beings with diabetes mellitus. Biochem Biophys Res Commun 76: 765–771
Silbert CK, Kleinman HK (1978) Studies of cultured human fibroblasts in diabetes mellitus: changes in heparan sulfate. Diabetes 28:61–65
Craig JW, Huang LS, Larner J (1980) Insulin stimulation of glycogen synthase activity in cultured human fibroblasts from diabetic and control subjects. Diabetologia 18:109–113
Rosenbloom AL, Rosenbloom EK (1978) Insulin-dependent childhood diabetes: normal viability of cultured fibroblasts. Diabetes 27: 338–341
Howard B, Fields R, Mott D, Savage P, Nagulesparan M, Bennett P (1980) Diabetes and cell growth: lack of differences in growth characteristics of fibroblasts from diabetic and non-diabetic Pima indians. Diabetes 29:119–124
Chait A, Bierman E, Albers J (1979) Low density lipoprotein receptor activity in fibroblasts cultured from diabetic donors. Diabetes 28:914–918
Goldstein S, Littlefield JW (1969) Effect of insulin on the conversion of glucose-C-14 to C-14-02 by nornal and diabetic fibroblasts in culture. Diabetes 18: 545–549
Zielke HR, Ozand PT, Tildon JT, Sevdalian DA, Cornblath M (1976) Growth of human diploid fibroblasts in the absence of glucose utilization. Proc Natl Acad Sci USA 73: 4110–4114
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Podskalny, J.M., Kahn, C.R. Insulin binding and activation of glycogen synthase in fibroblasts from type 1 (insulin-dependent) diabetic patients. Diabetologia 23, 431–435 (1982). https://doi.org/10.1007/BF00260957
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00260957