Summary
Insulin, proinsulin and glucagon extracted from lean rat pancreases were studied in radioimmunoassay, radioreceptorassay and bioassay systems. Extracted insulin behaved identically to a rat insulin used as a reference standard in radioimmunoassay. On the basis of its immunoreactivity, extracted insulin was slightly less potent (about 70%) than the rat standard insulin in competing with the binding of125I-insulin to rat liver membranes (radioreceptorassay) and in stimulating glucose oxidation by rat fat cells (bioassay). Extracted glucagon and a pork glucagon used as a reference standard were indistinguishable in two radioimmunoassay systems for glucagon, in competing with the binding of125I-glucagon to rat liver membranes (radioreceptorassay) and in stimulating adenylate cyclase in rat liver membranes (bioassay). Genetically obese rats (Zucker, “fatty”) were compared to their lean littermates with respect to insulin, proinsulin and glucagon extracted from their pancreases. Proinsulin represented the same proportion of total immunoreactive insulin in both types of rats. In the radioimmunoassays, the radioreceptorassays and the bioassays, insulin, proinsulin and glucagon from obese rats were indistinguishable from insulin, proinsulin and glucagon from lean rats. It is concluded that the pancreatic hormones of obese (“fatty”) rats possess the same immunoreactivity and biological potency as those of nonobese rats. This excludes the possibility that some alteration in the biological properties of pancreas insulin and/or glucagon of fatty rats could explain the metabolic abnormalities observed in this type of obesity.
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Berson, S.A., Yalow, R.S.: Insulin in blood and insulin antibodies. Amer. J. Med.40, 676–690 (1966)
Chance, R.E.: Chemical, physical, biological and immunological studies on porcine proinsulin and related peptides. In: Diabetes (eds. R.R. Rodriguez, J. Vallance-Owen), pp. 292–305. Amsterdam: Excerpta Medica, I. C. S. n∘ 231, 1 vol. 1971
Heding, L.G.: Immunological properties of pancreatic glucagon: antigenicity and antibody characteristics. In: Glucagon: molecular physiology, clinical and therapeutic implications, pp. 187–200. Oxford: Pergamon Press, 1 vol. 1972
Faloona, G.R.: The structure — function relationships of pancreatic glucagon. In: Glucagon: molecular physiology, clinical and therapeutic implications, pp. 201–204. Oxford: Pergamon Press, 1 vol. 1972
Freychet, P., Roth, J., Neville, D.M., Jr.: Monoiodoinsulin: demonstration of its biological activity and binding to fat cells and liver membranes. Biochem. biophys. Res. Commun.43, 400–408 (1971)
Freychet, P., Roth, J., Neville, D.M., Jr: Insulin receptors in the liver: Specific binding of125I-insulin to the plasma membrane and its relation to insulin bioactivity. Proc. nat. Acad. Sci. (Wash.)68, 1833–1837 (1971)
Rodbell, M., Krans, M.J., Pohl, S.L., Birnbaumer, L.: The glucagon sensitive adenyl cyclase system in plasma membranes of rat liver. III Binding of glucagon: method of assay and specificity. J. biol. Chem.246, 1861–1871 (1971)
Zucker, L.M., Zucker, T.G.: Fatty, a new mutation in the rat. J. Hered.52, 275–278 (1961)
Kenny, A.J.: Extractable glucagon of human pancreas. J. clin. Endocr.15, 1089–1105 (1955)
Rançon, F., Laburthe, M., Rosselin, G., Freychet, P.: Untractable hypoglycemia in an infant: studies on pancreas insulin and glucagon. Horm. Metab. Res.6, 443–447 (1974)
Yalow, R.S., Berson, S.A.: Immunoassay of endogenous plasma insulin in man. J. clin. Invest.39, 1157–1175 (1960)
Rosselin, G., Dolais, J.: Application de la méthode radio-immunologique au dosage de l'insuline humaine et au dosage de l'hormone folliculostimulante humaine (H FSH). Monographie ann. Sté Fse Biol. Clin. “Les Isotopes — Applications biocliniques”, Expansion Sci. Ed. pp. 189–217. 1. Vol. 1967
Unger, R.H., Eisentraut, A.M., Mac Call, M.S., Madison, L.L.: Glucagon antibodies and an immunoassay for glucagon. J. clin. Invest.40, 1280–1289 (1961)
Heding, L.G.: Radioimmunological determination of pancreatic and gut glucagon in plasma. Diabetologia7, 10–19 (1970)
Nottey, J.J., Rosselin, G.: Monoiodoglucagon: préparation, isolement, identification, contrôle radio-immunologique. C. R. Acad. Sci. (Paris)273, 2118–2121 (1971)
Rosselin, G., Assan, R., Yalow, R.S., Berson, S.A.: Separation of antibody — bound and unbound peptide hormones labeled with iodine 131 by talcum powder and precipitated silica. Nature (Lond.)212, 355–357 (1966)
Freychet, P.: The interactions of proinsulin with insulin receptors on the plasma membrane of the liver. J. clin. Invest.54, 1020–1031 (1974)
Freychet, P., Brandenburg, D., Wollmer, A.: Receptor-binding assay of chemically modified insulins. Diabetologia10, 1–5 (1974)
Bataille, D., Freychet, P., Kitabgi, P., Rosselin, G.: Gut glucagon: a common receptor site with pancreatic glucagon in liver cell plasma membranes FEBS Letters30, 215–218 (1973)
Neville, D.M., Jr: Isolation of an organ specific protein antigen from cell-surface membrane of rat liver. Biochim. biophys. Acta (Amst.)154, 540–552 (1968)
Freychet, P.: Insulin receptors. In: Methods in receptor research (ed. M. Blecher), New York: Marcel Dekker Inc. (in press, 1975)
Freychet, P., Kahn, R., Roth, J., Neville, D.M., Jr: Insulin interactions with liver plasma membranes. Independence of binding of the hormone and its degradation. J. biol. Chem.247, 3953–3961 (1972)
Desbuquois, B., Krug, F., Cuatrecasas, P.: Inhibitors of glucagon inactivation. Effect on glucagon-receptor interactions and glucagon — stimulated adenylate cyclase activity in liver cell membranes. Biochim. biophys. Acta (Amst.)343, 101–120 (1974)
Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the Folin phenol reagent. J. biol. Chem.193, 265–275 (1951)
Rosselin, G., Freychet, P.: Basal and hormone — stimulated adenylate cyclase in liver plasma membranes: Measurement by radioimmunoassay of cyclic AMP. Biochim. biophys. Acta (Amst.)304, 541–551 (1973)
Simon, J., Freychet, P., Rosselin, G.: Chicken insulin: radioimmunological characterization and enhanced activity in rat fat cells and liver plasma membranes. Endocrinology95, 1439–1449 (1974)
Rodbell, M.: Metabolism of isolated fat cells. I. Effects of hormones on glucose metabolism and lipolysis. J. biol. Chem.239, 375–380 (1964)
Jarrousse, C., Rançon, F., Rosselin, G.: Hormonogénèse périnatale de l'insuline et du glucagon chez le rat. C. R. Acad. Sci. (Paris)276, 585–588 (1973)
Kitabchi, A.: The biological and immunological properties of pork and beef insulin, proinsulin and connecting peptide. J. clin. Invest.49, 979–987 (1970)
Gliemann, J., Sorensen, H.H.: Assay of insulin-like activity by the isolated fat cell method. IV. The biological activity of proinsulin. Diabetologia6, 499–504 (1970)
Gammeltoft, S., Gliemann, J.: Binding and degradation of125I-insulin by isolated rat fat cells. Biochim. biophys. Acta (Amst.)320, 16–32 (1973)
Soeldner, J.J., Sawin, C.T., Silbert, C.K., Mitchell, M.L.: Differences in immunologic activity among preparations of human insulin. Metabolism23, 209–214 (1973)
Sundby, F., Markussen, J.: Isolation, crystallization and amino acid composition of rat glucagon. Horm. Metab. Res.3, 184–187 (1971)
Freychet, P., Laudat, M.H., Laudat, P., Rosselin, G., Kahn, C.R., Gorden, P., Roth, J.: Impairment of insulin binding to the fat cell plasma membrane in the obese hyperglycemie mouse. FEBS Letters25, 339–342 (1972)
Kahn, C.R., Neville, D.M., Jr, Roth, J.: Insulin — receptor interaction in the obese hyperglycémie mouse. A model of insulin resistance. J. biol. Chem.248, 244–250 (1973)
Shino, A., Matsuo, T., Iwatsuka, H., Suzuoki, Z.: Structural changes of pancreatic islets in genetically obese rats. Diabetologia9, 413–421 (1973)
Smith, L.F.: Species variation in the amino acid sequence of insulin. Amer. J. Med.40, 662–666 (1966)
Stauffacher, W., Lambert, A.E., Vecchio, D., Renold A.E.: Measurements of insulin activities in pancreas and serum of mice with spontaneous (“obese” and “New Zealand obese”) and induced (gold-thioglucose) obesity and hyperglycemia, with considerations on the pathogenesis of the spontaneous syndrome. Diabetologia3, 230–237 (1967)
Zucker, L.M.: Hereditary obesity in the rat associated with hyperlipemia. Ann. N. Y. Acad. Sci.131, 447–458 (1965)
Barry, N.S., Bray, G.A.: Plasma triglycerides in genetically obese rats. Metabolism18, 833–839 (1969)
Zucker, T.F., Zucker, L.M.: Hereditary obesity in the rat associated with high serum fat and cholesterol. Proc. Soc. exp. Biol. (N. Y.)110, 165–171 (1962)
Vaughan, M., Steinberg, D., Pittman, R.: On the interpretation of studies measuring uptake and esterification of I-14C palmitic acid by rat adipose tissue in vitro. Biochim. biophys. Acta (Amst.)84, 154–166 (1964)
Meikle, A.W., Klain, G.F., Hannon, J.P.: Inhibition of glucose oxidation and fatty acid synthesis in liver slices from fed, fasted and fasted-refed rats by glucagon epinephrine and cyclic adenosine-3′, 5′-monophosphate. Proc. Soc. exp. Biol. (N. Y.)143, 379–381 (1973)
Klain, G.J., Weiser, P.C.: Changes in hepatic fatty acid synthesis following glucagon injections in vivo. Biochem. biophys. Res. Commun.55, 76–83 (1973)
Eaton, R.P., Shade, D.S.: Glucagon resistance as a hormonal basis for endogenous hyperlipemia. Lancet1973 I, 973–974
Lemonnier, D., Aubert, R., Suquet, J.P., Rosselin, G.: Metabolism of genetically obese rats on normal or high-fat diet. Diabetologia10, 697–701 (1974)
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Laburthe, M., Rançon, F., Freychet, P. et al. Glucagon and insulin from lean rats and genetically obese fatty rats: Studies by radioimmunoassay, radioreceptorassay and bioassay. Diabetologia 11, 517–526 (1975). https://doi.org/10.1007/BF01222101
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DOI: https://doi.org/10.1007/BF01222101