Summary
Glutathione-insulin transhydrogenase catalyzes the inactivation of insulin by splitting the hormone into A and B chains. We have localized this enzyme immunocytochemically by light microscopy in the pancreas, kidney and liver of both lean and obese (ob/ob) mice and similarly in normal and streptozotocin-diabetic rats. Localization was achieved by an antibody to glutathione-insulin transhydrogenase using a peroxidase-antiperoxidase technique. In comparison with tissues from control animals, positive immunostaining for glutathione-insulin transhydrogenase was increased in the obese mouse but reduced in the diabetic rat. Different tissues showed considerable variation in the amount of glutathioneinsulin transhydrogenase which could be detected. In the pancreatic islets there was little or no evidence for the presence of the enzyme in peripheral cells. In the kidney, immunocytochemical staining was found only in the proximal tubules. In the liver there was a generalised distribution of the enzyme, but the greatest concentration was in the periportal region. These observations parallel the biochemical data relating to glutathione-insulin transhydrogenase, indicating that different amounts of insulin degrading activity exist in different regions of tissues.
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Beatens D, Stefan Y, Ravazzola M, Malaisse-Lagae F, Coleman DL, Orci L (1978) Alteration of islet cell populations in spontaneously diabetic mice. Diabetes 27: 1–7
Chandler ML, Varandani PT (1972) Insulin degradation. II. The wide-spread distribution of glutathione-insulin transhydrogenase in the tissues of the rat. Biochim Biophys Acta 286: 136–145
Chandler ML, Varandani PT (1974) Insulin degradation. IX. n the presence of glutathione-insulin transhydrogenase in human leukocytes. Diabetes 23: 232–239
Chandler ML, Varandani PT (1975) Kinetic analysis of the mechanism of insulin degradation by glutathione-insulin transhydrogenase (thiol: protein disulfide oxidoreductase). Biochemistry 14: 2107–2115
Cudworth AG, Barber HE (1975) The effect of hydrocortisone phosphate, methylpredinisolone and phenytoin on pancreatic insulin release and hepatic glutathione-insulin transhydrogenase activity in the rat. Eur J Pharmacol 31: 23–38
Dorn A, Kohnert K-D, Ansorge S, Zühlke H Arlt B, Lorenz D (1978) The immunohistochemical demonstration of the thiol: protein disulfide oxidoreductase (TPO) in pancreas and isolated Langerhans' islets: a light, fluorescent and electron microscopic study. Acta Histochem 63: 200–208
Graham RC, Karnovsky MJ (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14: 291–301
Herzberg L, Coleman DL (1977) Laboratory animals exhibiting obesity and diabetes syndromes. Metabolism 26: 59–99
Karakash C, Assimacopoulos-Jeannet F, Jeanrenaud B (1976) An anomaly of insulin removal in perfused livers of obese hyperglycemic (ob/ob) mice. J Clin Invest 57: 1117–1124
McCarroll AM, Buchanan KD (1973) Insulin clearance by the isolated perfused livers of insulin deficient rats. Diabetologia 9: 457–460
Petri W Jr, Rodriguez J, Pitot HC (1976) Environmental effects on glutathione-insulin transhydrogenase in rat liver. Proc Soc Exp Biol Med 152: 610–614
Phelps BH, Varandani PT (1977) Stimulation by insulin of cyclic AMP phosphodiesterase. Role of glutathione-insulin transhydrogenase. Biochem Biophys Res Commun 75: 302–310
Sacks H, Pimstone B, Waligora K, Peires L, Weinkove E, Saunders S (1977) Differences between insulin degradation by soluble fractions and insulin clearance by perfused livers of normal, protein-depleted, and starved rats. Diabetes 26: 956–966
Sternberger LA, Hardy PH, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody compley (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18: 315–333
Taylor CR (1978) Immunocytochemical methods in the study of lymphoma and related conditions. J Histochem Cytochem 26: 496–512
Thomas JH, Varandani PT (1979) Insulin degradation. XXV. Glutathione-insulin transhydrogenase activity of rat liver and kidney during the development of streptozotocin-diabetes. Biochim Biophys Acta 567: 88–95
Thomas JH, Wakefield SM, Jones RH (1973) The effect of insulin deficiency on the glutathione-insulin transhydrogenase activity of rat liver. Biochem Soc. Trans 1: 1179–1182
Uete T, Shimano N, Shimizu S, Morikawa M (1976) Autoregulatory system of insulin degradation in liver. II. Relationship between blood insulin levels and GSH-dependent insulin degrading activity in liver and blood. Metabolism 25: 375–384
Varandani PT (1973) Insulin degradation. IV. Sequential degradation of insulin by rat kidney, heart and skeletal muscle homogenates. Biochim Biophys Acta 295: 630–636
Varandani PT (1973) Insulin degradation. V. Unmasking of glutathione-insulin transhydrogenase in rat liver microsomal membrane. Biochim Biophys Acta 304: 642–659
Varandani PT (1973) Insulin degradation. VII. Sequential degradation of insulin by rat liver homogenates at physiological concentrations of insulin and in the absence of exogenous glutathione. Biochim Biophys Acta 320: 249–257
Varandani PT (1974) Insulin degradation in insulinoma: evidence for the occurrence of an inactive form of glutathione-insulin transhydrogenase and for the absence of insulin A and B chains degrading protease(s). Biochem Biophys Res Commun 60: 1119–1126
Varandani PT (1974) Insulin degradation. VI. Feedback control by insulin of liver glutathione-insulin transhydrogenase in rat. Diabetes 23: 117–125
Varandani PT (1976) The role of glutathione-insulin transhydrogenase in insulin degradation. In: Proc. IX Congress of the International Diabetes Federation (New Delhi, India). Excerpta Medica, Amsterdam, pp 213–223
Varandani PT (1978) Mechanistic and structural aspects of glutathione-insulin transhydrogenase (protein-disulfide interchange enzyme). In: Singer TP, Ondarza RN (eds) Mechanisms of oxidizing enzymes. Elsevier/North-Holland, New York, pp 29–42
Varandani PT, Darrow RM, Nafz MA (1977) Phospholipid composition of liver homogenates and microsomes of the lean and obese hyperglycemic (ob/ob) mouse. Proc Soc Exp Biol Med 156: 123–126
Varandani PT, Nafz MA (1972) Insulin degradation. I. Purification and properties of glutathione-insulin transhydrogenase of rat liver. Biochim Biophys Acta 286: 126–135
Varandani PT, Nafz MA (1976) Interaction of glutathione-insulin transhydrogenase (disulfide interchange enzyme) with phospholipids. Biochim Biophys Acta 438: 358–369
Varandani PT, Nafz MA (1976) Insulin degradation. XVI. Evidence for the sequential degradative pathway in isolated liver cells. Diabetes 25: 173–179
Varandani PT, Nafz MA (1976) Insulin degradation. XVIII. On the regulation of glutathione-insulin transhydrogenase in the hyperglycemic obese (ob/ob) mouse. Biochim Biophys Acta 451: 382–392
Varandani PT, Nafz MA, Chandler ML (1975) Interaction of insulin analogs, glucagon, growth hormone, vasopressin, oxytocin, and scrambled forms of ribonuclease and lysozyme with glutathione-insulin transhydrogenase (thiol: protein disulfide oxidoreductase): dependence upon conformation. Biochemstry 14: 2115–2120
Varandani PT, Nafz MA, Shroyer LA (1971) Glutathione-insulin transhydrogenase: evidence for a key role in insulin metabolism. Diabetes 20 [Suppl 1]: 342
Varandani PT, Raveed D, Nafz MA (1978) Insulin degradation XXIII. Distribution of glutathione-insulin transhydrogenase in isolated rat hepatocytes as studied by immuno-ferritin and electron microscopy. Biochim Biophys Acta 538: 343–353
Varandani PT, Shroyer LA, Nafz MA (1972) Sequential degradation of insulin by rat liver homogenates. Proc Natl Acad Sci USA 69: 1681–1684
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Taylor, C.A., Varandani, P.T. Insulin degradation. XXVIII. Immunocytochemical localization of glutathione-insulin transhydrogenase in the pancreas, kidney and liver of normal and streptozotocin-diabetic rats and of lean and obese (ob/ob) mice. Diabetologia 21, 464–469 (1981). https://doi.org/10.1007/BF00257787
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DOI: https://doi.org/10.1007/BF00257787