Insulin action and characterization of insulin receptors in rat luteal cells

The effect of insulin (Ins) on luteal cell function was assayed and Ins binding was characterized in isolated collagenase-dispersed rat luteal cells. Ins stimulated progesterone production at concentrations over 10-8 M, whereas human CG (hCG) produced maximal stimulation at 10- M. Ins had an additiv...

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Autores principales:	Ladenheim, Ruth Graciela, Tesone, Marta, Charreau, Eduardo Hernán
Publicado:	1984
Materias:	chorionic gonadotropin cyclic amp epidermal growth factor growth hormone insulin insulin i 125 insulin receptor luteinizing hormone progesterone prolactin radioisotope seric gonadotropin animal cell animal experiment drug administration drug binding drug efficacy drug interaction drug receptor binding endocrine system etiology female genital system luteal cell methodology nonhuman ovary pharmacokinetics progesterone h 3 rat steroidogenesis Animal Chorionic Gonadotropin Corpus Luteum Cyclic AMP Diabetes Mellitus, Insulin-Dependent Female Insulin Progesterone Rats Rats, Inbred Strains Receptor, Insulin Statistics Stimulation, Chemical Support, Non-U.S. Gov't Trypsin
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00137227_v115_n2_p752_Ladenheim http://hdl.handle.net/20.500.12110/paper_00137227_v115_n2_p752_Ladenheim
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paper:paper_00137227_v115_n2_p752_Ladenheim2023-06-08T14:36:04Z Insulin action and characterization of insulin receptors in rat luteal cells Ladenheim, Ruth Graciela Tesone, Marta Charreau, Eduardo Hernán chorionic gonadotropin cyclic amp epidermal growth factor growth hormone insulin insulin i 125 insulin receptor luteinizing hormone progesterone prolactin radioisotope seric gonadotropin animal cell animal experiment drug administration drug binding drug efficacy drug interaction drug receptor binding endocrine system etiology female genital system luteal cell methodology nonhuman ovary pharmacokinetics progesterone h 3 rat steroidogenesis Animal Chorionic Gonadotropin Corpus Luteum Cyclic AMP Diabetes Mellitus, Insulin-Dependent Female Insulin Progesterone Rats Rats, Inbred Strains Receptor, Insulin Statistics Stimulation, Chemical Support, Non-U.S. Gov't Trypsin The effect of insulin (Ins) on luteal cell function was assayed and Ins binding was characterized in isolated collagenase-dispersed rat luteal cells. Ins stimulated progesterone production at concentrations over 10-8 M, whereas human CG (hCG) produced maximal stimulation at 10- M. Ins had an additive effect when it was added to the luteal cells in the presence of hCG in concentrations that produced submaximal stimulation. At 10-9 M Ins the additive effect on hCG response became apparent (80% of maximal progesterone production). The maximal hCG response cannot be exceeded, even in the presence of high amounts of both hormones. hCG maximally stimulated cAMP production at 10-10 M, whereas Ins neither stimulated cAMP production nor enhanced hCG response. Ins binding to luteal cells attained highest values after 30 min of incubation at 20 C. A curvilinear Scatchard plot was obtained and it was analyzed using a two-binding site model. The affinity constant values were 2.1 × 108 M-1 and 5.2 × 106 M-1 and the number of binding sites were 35,000 and 900,000 per cell, respectively. Bound Ins was not displaceable by hCG, human GH, human LH, epidermal growth factor, or ovine PRL. A protein moiety was essential for the receptor activity, since after trypsin digestion marked loss of binding was verified. Subcellular fractionation was performed and the highest binding was observed in the 105,000 × g pellet fraction. This study demonstrates that Ins binds specifically to rat luteal cells and stimulates steroidogenesis, adding support for the hypothesis that insulin acts directly upon the ovary. © 1984 by The Endocrine Society. Fil:Ladenheim, R.G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Tesone, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Charreau, E.H. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1984 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00137227_v115_n2_p752_Ladenheim http://hdl.handle.net/20.500.12110/paper_00137227_v115_n2_p752_Ladenheim
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	chorionic gonadotropin cyclic amp epidermal growth factor growth hormone insulin insulin i 125 insulin receptor luteinizing hormone progesterone prolactin radioisotope seric gonadotropin animal cell animal experiment drug administration drug binding drug efficacy drug interaction drug receptor binding endocrine system etiology female genital system luteal cell methodology nonhuman ovary pharmacokinetics progesterone h 3 rat steroidogenesis Animal Chorionic Gonadotropin Corpus Luteum Cyclic AMP Diabetes Mellitus, Insulin-Dependent Female Insulin Progesterone Rats Rats, Inbred Strains Receptor, Insulin Statistics Stimulation, Chemical Support, Non-U.S. Gov't Trypsin
spellingShingle	chorionic gonadotropin cyclic amp epidermal growth factor growth hormone insulin insulin i 125 insulin receptor luteinizing hormone progesterone prolactin radioisotope seric gonadotropin animal cell animal experiment drug administration drug binding drug efficacy drug interaction drug receptor binding endocrine system etiology female genital system luteal cell methodology nonhuman ovary pharmacokinetics progesterone h 3 rat steroidogenesis Animal Chorionic Gonadotropin Corpus Luteum Cyclic AMP Diabetes Mellitus, Insulin-Dependent Female Insulin Progesterone Rats Rats, Inbred Strains Receptor, Insulin Statistics Stimulation, Chemical Support, Non-U.S. Gov't Trypsin Ladenheim, Ruth Graciela Tesone, Marta Charreau, Eduardo Hernán Insulin action and characterization of insulin receptors in rat luteal cells
topic_facet	chorionic gonadotropin cyclic amp epidermal growth factor growth hormone insulin insulin i 125 insulin receptor luteinizing hormone progesterone prolactin radioisotope seric gonadotropin animal cell animal experiment drug administration drug binding drug efficacy drug interaction drug receptor binding endocrine system etiology female genital system luteal cell methodology nonhuman ovary pharmacokinetics progesterone h 3 rat steroidogenesis Animal Chorionic Gonadotropin Corpus Luteum Cyclic AMP Diabetes Mellitus, Insulin-Dependent Female Insulin Progesterone Rats Rats, Inbred Strains Receptor, Insulin Statistics Stimulation, Chemical Support, Non-U.S. Gov't Trypsin
description	The effect of insulin (Ins) on luteal cell function was assayed and Ins binding was characterized in isolated collagenase-dispersed rat luteal cells. Ins stimulated progesterone production at concentrations over 10-8 M, whereas human CG (hCG) produced maximal stimulation at 10- M. Ins had an additive effect when it was added to the luteal cells in the presence of hCG in concentrations that produced submaximal stimulation. At 10-9 M Ins the additive effect on hCG response became apparent (80% of maximal progesterone production). The maximal hCG response cannot be exceeded, even in the presence of high amounts of both hormones. hCG maximally stimulated cAMP production at 10-10 M, whereas Ins neither stimulated cAMP production nor enhanced hCG response. Ins binding to luteal cells attained highest values after 30 min of incubation at 20 C. A curvilinear Scatchard plot was obtained and it was analyzed using a two-binding site model. The affinity constant values were 2.1 × 108 M-1 and 5.2 × 106 M-1 and the number of binding sites were 35,000 and 900,000 per cell, respectively. Bound Ins was not displaceable by hCG, human GH, human LH, epidermal growth factor, or ovine PRL. A protein moiety was essential for the receptor activity, since after trypsin digestion marked loss of binding was verified. Subcellular fractionation was performed and the highest binding was observed in the 105,000 × g pellet fraction. This study demonstrates that Ins binds specifically to rat luteal cells and stimulates steroidogenesis, adding support for the hypothesis that insulin acts directly upon the ovary. © 1984 by The Endocrine Society.
author	Ladenheim, Ruth Graciela Tesone, Marta Charreau, Eduardo Hernán
author_facet	Ladenheim, Ruth Graciela Tesone, Marta Charreau, Eduardo Hernán
author_sort	Ladenheim, Ruth Graciela
title	Insulin action and characterization of insulin receptors in rat luteal cells
title_short	Insulin action and characterization of insulin receptors in rat luteal cells
title_full	Insulin action and characterization of insulin receptors in rat luteal cells
title_fullStr	Insulin action and characterization of insulin receptors in rat luteal cells
title_full_unstemmed	Insulin action and characterization of insulin receptors in rat luteal cells
title_sort	insulin action and characterization of insulin receptors in rat luteal cells
publishDate	1984
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00137227_v115_n2_p752_Ladenheim http://hdl.handle.net/20.500.12110/paper_00137227_v115_n2_p752_Ladenheim
work_keys_str_mv	AT ladenheimruthgraciela insulinactionandcharacterizationofinsulinreceptorsinratlutealcells AT tesonemarta insulinactionandcharacterizationofinsulinreceptorsinratlutealcells AT charreaueduardohernan insulinactionandcharacterizationofinsulinreceptorsinratlutealcells
_version_	1768544532776878080

Insulin action and characterization of insulin receptors in rat luteal cells

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