Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells

The L-type Ca 2+ channel is the primary voltage-dependent Ca 2+ -influx pathway in many excitable and secretory cells, and direct phosphorylation by different kinases is one of the mechanisms involved in the regulation of its activity. The aim of this study was to evaluate the participation of Ser/T...

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Publicado:	2007
Materias:	Epidermal growth factor Phosphatases Protein kinase A Protein kinase C 4 (3 chloroanilino) 6,7 dimethoxyquinazoline calcium channel calcium channel L type calcium ion chelerythrine cyclic AMP dependent protein kinase epidermal growth factor epidermal growth factor receptor kinase inhibitor fura 2 acetoxymethyl ester genistein n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide nifedipine okadaic acid orthovanadic acid phorbol 13 acetate 12 myristate phosphatase phosphotransferase potassium chloride potassium ion protein kinase C protein kinase C inhibitor protein kinase p60 protein serine threonine kinase protein tyrosine kinase tyrosine kinase receptor vasculotropin voltage gated calcium channel animal cell article calcium cell level calcium mobilization calcium transport cell line cell membrane cell membrane depolarization cell proliferation cell strain GH3 controlled study enzyme activity enzymology gene expression hormone release hypophysis cell nonhuman priority journal protein synthesis rat spectrofluorometry Animals Calcium Calcium Channels, L-Type Carcinogens Cell Line Cell Line, Tumor Cyclic AMP-Dependent Protein Kinases Female Fluorescent Dyes Fura-2 Isoquinolines Membrane Potentials Pituitary Gland Pituitary Neoplasms Potassium Chloride Protein Kinase C Protein Kinase Inhibitors Protein Kinases Protein-Serine-Threonine Kinases Protein-Tyrosine Kinases Proto-Oncogene Proteins pp60(c-src) Rats Rats, Inbred WF Receptor, Epidermal Growth Factor Sulfonamides Tetradecanoylphorbol Acetate
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03636143_v293_n3_pC951_Vela http://hdl.handle.net/20.500.12110/paper_03636143_v293_n3_pC951_Vela
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_03636143_v293_n3_pC951_Vela
record_format	dspace
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Epidermal growth factor Phosphatases Protein kinase A Protein kinase C 4 (3 chloroanilino) 6,7 dimethoxyquinazoline calcium channel calcium channel L type calcium ion chelerythrine cyclic AMP dependent protein kinase epidermal growth factor epidermal growth factor receptor kinase inhibitor fura 2 acetoxymethyl ester genistein n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide nifedipine okadaic acid orthovanadic acid phorbol 13 acetate 12 myristate phosphatase phosphotransferase potassium chloride potassium ion protein kinase C protein kinase C inhibitor protein kinase p60 protein serine threonine kinase protein tyrosine kinase tyrosine kinase receptor vasculotropin voltage gated calcium channel animal cell article calcium cell level calcium mobilization calcium transport cell line cell membrane cell membrane depolarization cell proliferation cell strain GH3 controlled study enzyme activity enzymology gene expression hormone release hypophysis cell nonhuman priority journal protein synthesis rat spectrofluorometry Animals Calcium Calcium Channels, L-Type Carcinogens Cell Line Cell Line, Tumor Cyclic AMP-Dependent Protein Kinases Female Fluorescent Dyes Fura-2 Isoquinolines Membrane Potentials Pituitary Gland Pituitary Neoplasms Potassium Chloride Protein Kinase C Protein Kinase Inhibitors Protein Kinases Protein-Serine-Threonine Kinases Protein-Tyrosine Kinases Proto-Oncogene Proteins pp60(c-src) Rats Rats, Inbred WF Receptor, Epidermal Growth Factor Sulfonamides Tetradecanoylphorbol Acetate
spellingShingle	Epidermal growth factor Phosphatases Protein kinase A Protein kinase C 4 (3 chloroanilino) 6,7 dimethoxyquinazoline calcium channel calcium channel L type calcium ion chelerythrine cyclic AMP dependent protein kinase epidermal growth factor epidermal growth factor receptor kinase inhibitor fura 2 acetoxymethyl ester genistein n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide nifedipine okadaic acid orthovanadic acid phorbol 13 acetate 12 myristate phosphatase phosphotransferase potassium chloride potassium ion protein kinase C protein kinase C inhibitor protein kinase p60 protein serine threonine kinase protein tyrosine kinase tyrosine kinase receptor vasculotropin voltage gated calcium channel animal cell article calcium cell level calcium mobilization calcium transport cell line cell membrane cell membrane depolarization cell proliferation cell strain GH3 controlled study enzyme activity enzymology gene expression hormone release hypophysis cell nonhuman priority journal protein synthesis rat spectrofluorometry Animals Calcium Calcium Channels, L-Type Carcinogens Cell Line Cell Line, Tumor Cyclic AMP-Dependent Protein Kinases Female Fluorescent Dyes Fura-2 Isoquinolines Membrane Potentials Pituitary Gland Pituitary Neoplasms Potassium Chloride Protein Kinase C Protein Kinase Inhibitors Protein Kinases Protein-Serine-Threonine Kinases Protein-Tyrosine Kinases Proto-Oncogene Proteins pp60(c-src) Rats Rats, Inbred WF Receptor, Epidermal Growth Factor Sulfonamides Tetradecanoylphorbol Acetate Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells
topic_facet	Epidermal growth factor Phosphatases Protein kinase A Protein kinase C 4 (3 chloroanilino) 6,7 dimethoxyquinazoline calcium channel calcium channel L type calcium ion chelerythrine cyclic AMP dependent protein kinase epidermal growth factor epidermal growth factor receptor kinase inhibitor fura 2 acetoxymethyl ester genistein n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide nifedipine okadaic acid orthovanadic acid phorbol 13 acetate 12 myristate phosphatase phosphotransferase potassium chloride potassium ion protein kinase C protein kinase C inhibitor protein kinase p60 protein serine threonine kinase protein tyrosine kinase tyrosine kinase receptor vasculotropin voltage gated calcium channel animal cell article calcium cell level calcium mobilization calcium transport cell line cell membrane cell membrane depolarization cell proliferation cell strain GH3 controlled study enzyme activity enzymology gene expression hormone release hypophysis cell nonhuman priority journal protein synthesis rat spectrofluorometry Animals Calcium Calcium Channels, L-Type Carcinogens Cell Line Cell Line, Tumor Cyclic AMP-Dependent Protein Kinases Female Fluorescent Dyes Fura-2 Isoquinolines Membrane Potentials Pituitary Gland Pituitary Neoplasms Potassium Chloride Protein Kinase C Protein Kinase Inhibitors Protein Kinases Protein-Serine-Threonine Kinases Protein-Tyrosine Kinases Proto-Oncogene Proteins pp60(c-src) Rats Rats, Inbred WF Receptor, Epidermal Growth Factor Sulfonamides Tetradecanoylphorbol Acetate
description	The L-type Ca 2+ channel is the primary voltage-dependent Ca 2+ -influx pathway in many excitable and secretory cells, and direct phosphorylation by different kinases is one of the mechanisms involved in the regulation of its activity. The aim of this study was to evaluate the participation of Ser/Thr kinases and tyrosine kinases (TKs) in depolarization-induced Ca 2+ influx in the endocrine somatomammotrope cell line GH3. Intracellular Ca 2+ concentration ([Ca 2+ ] i ) was measured using a spectrofluorometric method with fura 2-AM, and 12.5 mM KCl (K + ) was used as a depolarization stimulus. K + induced an abrupt spike (peak) in [Ca 2+ ] i that was abolished in the presence of nifedipine, showing that K + enhances [Ca 2+ ] i , preferably activating L-type Ca 2+ channels. H89, a selective PKA inhibitor, significantly reduced depolarization-induced Ca 2+ mobilization in a concentration-related manner when it was applied before or after K + , and okadaic acid, an inhibitor of Ser/Thr phosphatases, which has been shown to regulate PKA-stimulated L-type Ca 2+ channels, increased K + -induced Ca 2+ entry. When PKC was activated by PMA, the K + -evoked peak in [Ca 2+ ] i , as well as the plateau phase, was significantly reduced, and chelerythrine (a PKC inhibitor) potentiated the K + -induced increase in [Ca 2+ ] i , indicating an inhibitory role of PKC in voltage-dependent Ca 2+ channel (VDCC) activity. Genistein, a TK inhibitor, reduced the K + -evoked increase in [Ca 2+ ] i , but, unexpectedly, the tyrosine phosphatase inhibitor orthovanadate reduced not only basal Ca 2+ levels but, also, Ca 2+ influx during the plateau phase. Both results suggest that different TKs may act differentially on VDCC activation. Activation of receptor TKs with epidermal growth factor (EGF) or vascular endothelial growth factor potentiated K + -induced Ca 2+ influx, and AG-1478 (an EGF receptor inhibitor) decreased it. However, inhibition of the non-receptor TK pp60 c-Src enhanced K + -induced Ca 2+ influx. The present study strongly demonstrates that a complex equilibrium among different kinases and phosphatases regulates VDCC activity in the pituitary cell line GH3: PKA and receptor TKs, such as vascular endothelial growth factor receptor and EGF receptor, enhance depolarization-induced Ca 2+ influx, whereas PKC and c-Src have an inhibitory effect. These kinases modulate membrane depolarization and may therefore participate in the regulation of a plethora of intracellular processes, such as hormone secretion, gene expression, protein synthesis, and cell proliferation, in pituitary cells. Copyright © 2007 the American Physiological Society.
title	Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells
title_short	Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells
title_full	Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells
title_fullStr	Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells
title_full_unstemmed	Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells
title_sort	different kinases regulate activation of voltage-dependent calcium channels by depolarization in gh3 cells
publishDate	2007
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03636143_v293_n3_pC951_Vela http://hdl.handle.net/20.500.12110/paper_03636143_v293_n3_pC951_Vela
_version_	1768544687689302016
spelling	paper:paper_03636143_v293_n3_pC951_Vela2023-06-08T15:35:29Z Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells Epidermal growth factor Phosphatases Protein kinase A Protein kinase C 4 (3 chloroanilino) 6,7 dimethoxyquinazoline calcium channel calcium channel L type calcium ion chelerythrine cyclic AMP dependent protein kinase epidermal growth factor epidermal growth factor receptor kinase inhibitor fura 2 acetoxymethyl ester genistein n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide nifedipine okadaic acid orthovanadic acid phorbol 13 acetate 12 myristate phosphatase phosphotransferase potassium chloride potassium ion protein kinase C protein kinase C inhibitor protein kinase p60 protein serine threonine kinase protein tyrosine kinase tyrosine kinase receptor vasculotropin voltage gated calcium channel animal cell article calcium cell level calcium mobilization calcium transport cell line cell membrane cell membrane depolarization cell proliferation cell strain GH3 controlled study enzyme activity enzymology gene expression hormone release hypophysis cell nonhuman priority journal protein synthesis rat spectrofluorometry Animals Calcium Calcium Channels, L-Type Carcinogens Cell Line Cell Line, Tumor Cyclic AMP-Dependent Protein Kinases Female Fluorescent Dyes Fura-2 Isoquinolines Membrane Potentials Pituitary Gland Pituitary Neoplasms Potassium Chloride Protein Kinase C Protein Kinase Inhibitors Protein Kinases Protein-Serine-Threonine Kinases Protein-Tyrosine Kinases Proto-Oncogene Proteins pp60(c-src) Rats Rats, Inbred WF Receptor, Epidermal Growth Factor Sulfonamides Tetradecanoylphorbol Acetate The L-type Ca 2+ channel is the primary voltage-dependent Ca 2+ -influx pathway in many excitable and secretory cells, and direct phosphorylation by different kinases is one of the mechanisms involved in the regulation of its activity. The aim of this study was to evaluate the participation of Ser/Thr kinases and tyrosine kinases (TKs) in depolarization-induced Ca 2+ influx in the endocrine somatomammotrope cell line GH3. Intracellular Ca 2+ concentration ([Ca 2+ ] i ) was measured using a spectrofluorometric method with fura 2-AM, and 12.5 mM KCl (K + ) was used as a depolarization stimulus. K + induced an abrupt spike (peak) in [Ca 2+ ] i that was abolished in the presence of nifedipine, showing that K + enhances [Ca 2+ ] i , preferably activating L-type Ca 2+ channels. H89, a selective PKA inhibitor, significantly reduced depolarization-induced Ca 2+ mobilization in a concentration-related manner when it was applied before or after K + , and okadaic acid, an inhibitor of Ser/Thr phosphatases, which has been shown to regulate PKA-stimulated L-type Ca 2+ channels, increased K + -induced Ca 2+ entry. When PKC was activated by PMA, the K + -evoked peak in [Ca 2+ ] i , as well as the plateau phase, was significantly reduced, and chelerythrine (a PKC inhibitor) potentiated the K + -induced increase in [Ca 2+ ] i , indicating an inhibitory role of PKC in voltage-dependent Ca 2+ channel (VDCC) activity. Genistein, a TK inhibitor, reduced the K + -evoked increase in [Ca 2+ ] i , but, unexpectedly, the tyrosine phosphatase inhibitor orthovanadate reduced not only basal Ca 2+ levels but, also, Ca 2+ influx during the plateau phase. Both results suggest that different TKs may act differentially on VDCC activation. Activation of receptor TKs with epidermal growth factor (EGF) or vascular endothelial growth factor potentiated K + -induced Ca 2+ influx, and AG-1478 (an EGF receptor inhibitor) decreased it. However, inhibition of the non-receptor TK pp60 c-Src enhanced K + -induced Ca 2+ influx. The present study strongly demonstrates that a complex equilibrium among different kinases and phosphatases regulates VDCC activity in the pituitary cell line GH3: PKA and receptor TKs, such as vascular endothelial growth factor receptor and EGF receptor, enhance depolarization-induced Ca 2+ influx, whereas PKC and c-Src have an inhibitory effect. These kinases modulate membrane depolarization and may therefore participate in the regulation of a plethora of intracellular processes, such as hormone secretion, gene expression, protein synthesis, and cell proliferation, in pituitary cells. Copyright © 2007 the American Physiological Society. 2007 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03636143_v293_n3_pC951_Vela http://hdl.handle.net/20.500.12110/paper_03636143_v293_n3_pC951_Vela

Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells

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