Insulin Receptors in Human Circulating Cells and Fibroblasts

Gavin, James R.; Roth, Jesse; Jen, Phyllis; Freychet, P

doi:10.1073/pnas.69.3.747

Cited by 209 publications

(82 citation statements)

References 25 publications

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“…Explants were cultured in BGJB medium for 18 hr and then transferred to a Tris-buffered Ringer's solution (9) with 2% albumin at pH 7.5, containing 1 ng/ml of [125 ]insulin plus various concentrations of unlabeled insulin or other additions. At the end of the experiment, explants were rinsed in the buffer (40) and the 125I bound was measured.…”

Section: Methodsmentioning

confidence: 99%

Regulation of Hepatic Glycogen Synthesis During Fetal Development: Roles of Hydrocortisone, Insulin, and Insulin Receptors

Eisen¹,

Goldfine

Glinsmann³

1973

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

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In fetal rat liver in utero, an increase in glycogen and the glycogen synthetic enzyme glycogen synthetase (EC 2.4.1.11) occurs between gestational days 17 and 19. We used an organ culture system for finding the stimulus for increased enzyme activity and defining the relationship of this increase to glycogen synthesis. In fetal-liver explants from an earlier period (gestational day 16), hydrocortisone causes an elevation in total glycogen synthetase activity. This effect, which can be blocked by actinomycin D, is strikingly similar in time course and magnitude to the normal increase in utero. However, in order for glycogen synthesis to proceed after hydrocortisone increases glycogen synthetase levels, insulin is required. Unlike hydrocortisone, insulin does not increase total glycogen synthetase; it appears to act by converting the b or phospho form of synthetase to the a or dephospho form. Insulin alone does not stimulate glycogen synthesis in explants from 16-day fetal liver, although no defect in insulin binding was demonstrable. These findings support the hypothesis that the increase in liver-glycogen synthesis during the last trimester requires glucocorticoids for the developmental induction of glycogen synthetase and insulin for activation of the enzyme.The glycogen content of fetal rat liver increases greatly during the last few days of gestation (1). Experiments in utero have shown that adrenal corticosteroids can increase glycogen in fetal liver as early as the 15th day of gestation (2). Experiments in vitro with term (gestational day 21) fetal-liver explants in organ culture have shown that insulin, but not hydrocortisone, stimulates glycogen synthesis at the end of gestation (3, 4). We postulated that this difference in the action of hydrocortisone in utero and in organ culture may be explained on the basis of differences in gestational age. The major developmental increase in glycogen and the glycogen synthetic enzyme glycogen synthetase (EC 2.4.1.11) occurs between the 17th and 19th days of fetal life in the rat (5). It is possible that the effect of hydrocortisone may be fully expressed in term tissue. We therefore studied the action of hydrocortisone and insulin on glycogen metabolism in fetal rat liver explants from day 16 of gestation. At this stage of gestation, hydrocortisone caused an increase in total glycogen synthetase (a+b) that was strikingly similar to the normal induction of the enzyme in utero. Glycogen synthesis, however, required insulin. Unlike glucocorticoids, insulin did not increase total glycogen synthetase (a+b), but appeared to convert the b or phospho form of synthetase to the a or dephospho form. In term fetal liver, total glycogen synthetase had already reached adult levels and was not further increased by hydrocortisone, whereas insulin increased synthetase a and stimulated glycogen synthesis.The two hormones had distinctly different effects on glycogen synthetase. Glucorticoids stimulated the developmental induction of total glycogen synthetase (a+b), while insul...

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Section: Methodsmentioning

confidence: 99%

Regulation of Hepatic Glycogen Synthesis During Fetal Development: Roles of Hydrocortisone, Insulin, and Insulin Receptors

Eisen¹,

Goldfine

Glinsmann³

1973

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

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“…Although insulin receptors have been identified for fat, liver, skeletal and cardiac muscle, fibroblasts and monocytes [1][2][3][4][5][6][7][8] , quiescent rat splenic T lymphocytes do not bear such receptors. However, insulin receptors do emerge on T lymphocytes after concanavalin A treatment or allogeneic skin grafting 9,10 .…”

mentioning

confidence: 99%

Specific insulin binding site on T and B lymphocytes as a marker of cell activation

Helderman

Strom

1978

Nature

167

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Attempts to identify an easily measured cell surface marker that would detect activation of T or B lymphocytes in a variety of species have been unsuccessful. Although insulin receptors have been identified for fat, liver, skeletal and cardiac muscle, fibroblasts and monocytes [1][2][3][4][5][6][7][8] , quiescent rat splenic T lymphocytes do not bear such receptors. However, insulin receptors do emerge on T lymphocytes after concanavalin A treatment or allogeneic skin grafting 9,10 . Following our work on characterising a lymphocyte insulin receptor which participates in the modulation of immune response 11 , we have examined and report here the possibility that the emergence of a membrane-bound receptor for the peptide hormone insulin may be a universal marker for activated T and B lymphocytes.Rat splenic lymphocyte cell suspensions were established in RPMI-1640 media buffered with 5 mM HEPES and fortified with 5% sterile, heat-inactivated fetal calf serum (designated 'enriched' media). Leukocytes were collected from a Ficoll-Hypaque gradient. The cells were fractionated on nylon wool columns yielding T-enriched, nylon woolnonadherent cells and B-enriched, nylon wool-adherent cells. Macrophages were removed from all final cell suspensions by removing the cells that phagocytose iron particles with magnetic attraction. Insulin binding assays were carried out in suspensions containing 10 7 cells per ml in Hank's balanced salt solution enriched with 0.1% bovine serum albumin for analysis of receptor binding or the 'enriched' media for analysis of thymidine incorporation. Such purification techniques were evaluated using cell surface markers for the Fc receptor, the complement receptor, surface immunoglobulin, anti-θ determinants, and acridine orange and p-rosaniline staining characteristics.The insulin receptor was measured by the use of a modification of the insulin binding assay described by Gammeltoft and Gliemann 12 which has been extensively described 10 . In brief, the assay involves a competitive binding reaction between radiolabelled insulin and unlabelled monocomponent porcine insulin. By such competition, both total and nonspecific isotopic binding may be measured, and specific (receptor) binding can be calculated. Specific binding is 50-60% of the total binding and is similar to that of the fibroblast. Final separation of cell-bound from free insulin was accomplished over oil with virtually no contamination of the cell fraction by free insulin and recovery of 93% of cells for analysis. Lymphocyte cultures were also evaluated for the intracellular accumulation of tritiated thymidine after a four-hour pulse of 10 6 cells in flat-bottom microtitre plates (1 μ Ci per well).© 1978 Nature Publishing Group * To whom reprint requests should be addressed at the Texas Health Science Center. NIH Public Access Author ManuscriptNature. Author manuscript; available in PMC 2014 January 07. Published in final edited form as:Nature. 1978 July 6; 274(5666): 62-63. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Au...

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“…The magnitude of this response depends on the hormone concentration, the receptor concentration and the affinity of the receptor, whereby alterations in any one of these can alter the biological response. Insulin receptors have been defined in human mononuclear cells (4)(5)(6)(7)(8) adipocytes (2,5), placental cells (9,10), cultured lymphocytes (l 1), hepatocytes (12), cultured fibroblasts (11), myocytes (13), gjanulocytes (14), reticulocytes (15) and erythrocytes (11,.…”

Section: Introductionmentioning

confidence: 99%

Insulin Binding to Erythrocytes from Pregnant, Postpartum, Follicular and Luteal States

Dwenger¹,

Mitzkat²,

Holle³

et al. 1982

CCLM

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By using a modified Scatchard analysis, statistically significant differences were observed between the receptor affinities of the groups A aiid D, B and D, A and C.The receptor affinities and concentrations were not significantly different between the follicular and the luteal phases. From the data, no inverse correlation between the plasma insulin concentration and receptor binding was seen, i.e. the phenömenon of downiegulation pf insulin receptor concentration with hyperinsulinaemia seemed not to apply to erythrocytes.The present results suggest that insulin binding to erythrocytes is mpdulated preferably or even exclusively by an alteration of receptor affinity and that short-term chariges in insulin binding to erythroeytes are not caused by an alterätion of receptor concentration. Vergleich der Bindung von Insulin an Erythrocyten während und nach Schwangerschaft und in der Proliferationsund der Sekretionsphase des Menstruations-Cyclus

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Insulin Receptors in Human Circulating Cells and Fibroblasts

Cited by 209 publications

References 25 publications

Regulation of Hepatic Glycogen Synthesis During Fetal Development: Roles of Hydrocortisone, Insulin, and Insulin Receptors

Regulation of Hepatic Glycogen Synthesis During Fetal Development: Roles of Hydrocortisone, Insulin, and Insulin Receptors

Specific insulin binding site on T and B lymphocytes as a marker of cell activation

Insulin Binding to Erythrocytes from Pregnant, Postpartum, Follicular and Luteal States

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