Hormone receptors 7. Characteristics of insulin receptors in a new line of cloned neonatal rat hepatocytes

Petersen, B. Nüchel; Beckner, Suzanne K.; Blecher, Melvin

doi:10.1016/0304-4165(78)90377-x

Cited by 46 publications

(7 citation statements)

References 26 publications

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“…The effect of chronic treatment ofcells with insulin on cell surface insulin receptor level is well documented. This phenomenon, known as ligand-induced receptor down-regulation, has been demonstrated in vivo with circulating monocytes (1) and fat cells (2) from obese humans and with cultured human IM-9 lymphocytes (3-6), rat hepatocytes (7), and chicken hepatocytes (8). The reciprocal effect-i.e., an increase in cell!…”

mentioning

confidence: 99%

Control of insulin receptor level in 3T3 cells: effect of insulin-induced down-regulation and dexamethasone-induced up-regulation on rate of receptor inactivation.

Knutson¹,

Ronnett²,

Lane³

1982

Proc. Natl. Acad. Sci. U.S.A.

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Chronic exposure of 3T3 mouse fibroblasts to insulin or to the glucocorticoid dexamethasone induces down-regulation and up-regulation, respectively, of cell-surface and total cellular insulin binding capacity. Both processes are reversed upon withdrawal of the inducer. Scatchard analysis of insulin binding for receptors in the down-and up-regulated states indicates that the changes in binding capacity result primarily from alterations in insulin receptor level. That these alterations in total receptor level are due to changes in cell-surface receptor level is indicated by the fact that the level of trypsin-insensitive, presumably intracellular, insulin binding sites does not change appreciably upon down-and up-regulation. The effects ofinsulin-induced down-regulation and dexamethasone-induced up-regulation on the rates of insulin receptor synthesis and~decay were assessed by the heavyisotope density-shift technique. Cells were shifted to medium containing heavy (2H, '3C, '5N) amino acids and, at various times after the shift, light and heavy receptors solubilized from total cellular membranes were resolved by isopycnic banding on density gradients and then quantitated. It was demonstrated that the insulinand dexamethasone-induced alterations in insulin receptor level were due entirely to changes in the rate constant for receptor inactivation. The decrease in the first-order rate constant for receptor decay caused by dexamethasone is unexpected in view of the known action of steroid hormones in the induction of the synthesis of specific proteins.The insulin receptor is a dynamic cellular protein whose equilibrium concentration in the plasma membrane can be altered by physiological perturbants. The effect of chronic treatment ofcells with insulin on cell surface insulin receptor level is well documented. This phenomenon, known as ligand-induced receptor down-regulation, has been demonstrated in vivo with circulating monocytes (1) and fat cells (2) from obese humans and with cultured human IM-9 lymphocytes (3-6), rat hepatocytes (7), and chicken hepatocytes (8). The reciprocal effect-i.e., an increase in cell! surface insulin binding capacity-is observed upon removal ofinsulin from a down-regulated system (3) or by induction of differentiation of 3T3-Ll preadipocytes into adipocytes (9-11).Changes in the level or affinity ofhormone receptors can also be induced by heterologous hormones (6). Glucocorticoids have been shown to affect cellular levels of prolactin receptors (12), epidermal growth factor receptors (13), and insulin, receptors (14-18). It was of interest, therefore, to determine the mechanism by which glucocorticoids induce an increased insulin binding capacity of cells.In this report the effects of two antagonistic perturbants of insulin receptor level in 3T3 cells are described: the effect of insulin, which causes down-regulation, and that of dexamethasone, which causes up-regulation ofinsulin receptor level. The heavy isotope density-shift technique (8,19,20) was employed to determine whether...

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mentioning

confidence: 99%

Control of insulin receptor level in 3T3 cells: effect of insulin-induced down-regulation and dexamethasone-induced up-regulation on rate of receptor inactivation.

Knutson¹,

Ronnett²,

Lane³

1982

Proc. Natl. Acad. Sci. U.S.A.

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“…Normal (70-150 population doublings) and transformed (300-400 population doublings) RL-PR-C hepatocytes were cultured as described (8,9). The criteria for spontaneous transformation were those of Schaeffer et al (6,7,12 The publication costs of this article were defrayed in part by page charge payment.…”

Section: Methodsmentioning

confidence: 99%

“…These cells represent a unique model system for investigating hormonal regulation of adenylate cyclase. For well over a year (over 200 population doublings) they remain stable culturally and karyotypically, during which time they display a wide variety of liver functions, including the production of many specific liver enzymes and structural proteins (7) and responsiveness to insulin (8), cholera toxin (9), catecholamines (10), and glucagon (11). After this stable periodi.e., between 200 and 300 population doublings-the cells spontaneously transform and acquire the cultural and chromosomal characteristics of cells transformed by artificial means, including loss of contact inhibition and tumorigenicity in isogenic animals (12), and also become insensitive to glucagon (11).…”

mentioning

confidence: 99%

Restoration of glucagon responsiveness in spontaneously transformed rat hepatocytes (RL-PR-C) by fusion with normal progenitor cells and rat liver plasma membranes.

Reilly¹,

Blecher²

1981

Proc. Natl. Acad. Sci. U.S.A.

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“…Down regulation has been demon-strated in several types ofinsulin-responsive cells, including rat adipocytes (6)(7)(8)(9)(10)(11)(12)(13). The decrease in insulin binding capacity was -50-60% and occurred half-maximally within 2-3 h at 37°C at high concentrations of insulin (reviewed in 14).…”

Section: Introductionmentioning

confidence: 99%

Desensitization of the insulin receptor by antireceptor antibodies in vivo is blocked by treatment of mice with beta-adrenergic agonists.

Elias

Rapoport²,

Cohen³

et al. 1988

J. Clin. Invest.

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In previous studies we reported that immunization of mice with ungulate insulins induced the development of antiinsulin antibodies, which include an idiotype that appeared to recognize the part of the insulin molecule recognized by the hormone receptor. The antiinsulin antibodies of this idiotype were replaced spontaneously by antiidiotypic antibodies. The antiidiotypic antibodies, which persisted for about 14 d, mimicked insulin and functioned as antibodies to the insulin receptor. They induced down regulation, desensitization and refractoriness of the insulin receptor and disturbances in glucose homeostasis in vivo (Shechter, Y., D. Elias, R. Maron, and I. R. Cohen., 1984; Elias, D., R. Maron, I. R. Cohen, and Y. Shechter. 1984, J. BioL Chem. 259:6411-6419). We now report that effects of the antiidiotypic antibodies on the insulin receptor effector system can be modified pharmacologically.Administration of the fl-adrenergic agonist isoproterenol during the period of insulin resistance (days 26-40 after primary immunization), largely restored fat cell responsiveness to insulin, and eliminated the appearance of fasting hyperglycemia. This restoration appeared to be caused by inhibition of both insulin receptor desensitization and refractoriness. In contrast, down regulation of insulin receptors was not reversed by isoproterenol treatment in vivo. The effects of treatment with isoproterenol persisted for 24 d after termination of treatment. The j-antagonist, propranolol and more so, the #I.-antagonist metoprolol, specifically blocked the effect of isoproterenol at a molar ratio of 3-10:1. Oral administration of the cAMP phosphodiesterase inhibitor, aminophylline, was also effective in inhibiting the development of desensitization in fat cells. These results indicate that treatment with 01-adrenergic agonists in vivo, or other agents that elevate cellular cAMP levels, can inhibit the development of the "postbinding" defects induced by insulin-mimicking, antireceptor antibodies. These observations have both basic and clinical implications.

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Hormone receptors 7. Characteristics of insulin receptors in a new line of cloned neonatal rat hepatocytes

Cited by 46 publications

References 26 publications

Control of insulin receptor level in 3T3 cells: effect of insulin-induced down-regulation and dexamethasone-induced up-regulation on rate of receptor inactivation.

Control of insulin receptor level in 3T3 cells: effect of insulin-induced down-regulation and dexamethasone-induced up-regulation on rate of receptor inactivation.

Restoration of glucagon responsiveness in spontaneously transformed rat hepatocytes (RL-PR-C) by fusion with normal progenitor cells and rat liver plasma membranes.

Desensitization of the insulin receptor by antireceptor antibodies in vivo is blocked by treatment of mice with beta-adrenergic agonists.

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