Regulation of Ca<inf>V</inf>3.1 channels by glucocorticoids

Traudy Avila; Oscar Hernández-Hernández; Angélica Almanza; Mario Bermúdez De León; Mercedes Urban; Enrique Soto; Bulmaro Cisneros; Ricardo Felix

doi:10.1007/s10571-009-9422-2

Regulation of Ca<inf>V</inf>3.1 channels by glucocorticoids

Traudy Avila, Oscar Hernández-Hernández, Angélica Almanza, Mario Bermúdez De León, Mercedes Urban, Enrique Soto, Bulmaro Cisneros, Ricardo Felix

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

The activity of low voltage-activated Ca ²⁺ (Ca _V3) channels is tightly coupled to neurotransmitter and hormone secretion. Previous studies have shown that Ca _V3 channels are regulated by glucocorticoids (GCs), though the mechanism underlying channel regulation remains unclear. Here, using the pituitary GH ₃ cell line as a model, we investigated whether Ca _V3 channel expression is under the control of GCs, and if their actions are mediated by transcriptional and/or post-transcriptional mechanisms. RT-PCR and western blot analyses showed that Ca _V3.1 but not Ca _V3.2 and Ca _V3.3 channels is expressed in the GH ₃ cells, and patch clamp recordings confirmed that Ca ²⁺ currents through low voltage-activated channels were decreased after chronic treatment with GCs. Consistent with this, total plasma membrane expression of Ca _V3.1 protein as analyzed by cell-surface biotinylation assays and semi-quantitative western blotting was also down-regulated, while quantitative real-time RT-PCR analysis revealed a significant decrease of Ca _V3.1 mRNA expression in the treated cells. In contrast, patch-clamp recordings on HEK-293 cells stably expressing recombinant Ca _V3.1 channels showed that Ca ²⁺ currents were not affected by GC treatment. These results suggest that decreased transcription is a likely mechanism to explain the inhibitory actions of GCs on the functional expression of native Ca _V3.1 channels.

Original language	English
Pages (from-to)	1265-1273
Number of pages	9
Journal	Cellular and Molecular Neurobiology
Volume	29
Issue number	8
DOIs	https://doi.org/10.1007/s10571-009-9422-2
Publication status	Published - 1 Dec 2009
Externally published	Yes

Bibliographical note

Funding Information:
Acknowledgments This work was supported by funds from Cona-cyt to RF. We thank Drs. M. E. Mendoza (Cinvestav-IPN, Mexico) and J. C. Gomora (IFC-UNAM, Mexico) for the generous gift of the cell lines and Dr. M. Hernandez (Cinvestav-IPN, Mexico) and D. Mornet (INSERM ERI 25 Muscle et Pathologies, France) for the anti-actin and anti-β-dystroglycan antibodies, respectively. We are also indebted to J. Arikkath for critically reviewing the manuscript.

Copyright:
Copyright 2010 Elsevier B.V., All rights reserved.

All Science Journal Classification (ASJC) codes

Cellular and Molecular Neuroscience
Cell Biology

Access to Document

10.1007/s10571-009-9422-2

https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=77049109229&origin=inward

Cite this

@article{2c090a2997674f9a9622ee1e430829b2,

title = "Regulation of CaV3.1 channels by glucocorticoids",

abstract = "The activity of low voltage-activated Ca 2+ (Ca V3) channels is tightly coupled to neurotransmitter and hormone secretion. Previous studies have shown that Ca V3 channels are regulated by glucocorticoids (GCs), though the mechanism underlying channel regulation remains unclear. Here, using the pituitary GH 3 cell line as a model, we investigated whether Ca V3 channel expression is under the control of GCs, and if their actions are mediated by transcriptional and/or post-transcriptional mechanisms. RT-PCR and western blot analyses showed that Ca V3.1 but not Ca V3.2 and Ca V3.3 channels is expressed in the GH 3 cells, and patch clamp recordings confirmed that Ca 2+ currents through low voltage-activated channels were decreased after chronic treatment with GCs. Consistent with this, total plasma membrane expression of Ca V3.1 protein as analyzed by cell-surface biotinylation assays and semi-quantitative western blotting was also down-regulated, while quantitative real-time RT-PCR analysis revealed a significant decrease of Ca V3.1 mRNA expression in the treated cells. In contrast, patch-clamp recordings on HEK-293 cells stably expressing recombinant Ca V3.1 channels showed that Ca 2+ currents were not affected by GC treatment. These results suggest that decreased transcription is a likely mechanism to explain the inhibitory actions of GCs on the functional expression of native Ca V3.1 channels.",

author = "Traudy Avila and Oscar Hern{\'a}ndez-Hern{\'a}ndez and Ang{\'e}lica Almanza and {Berm{\'u}dez De Le{\'o}n}, Mario and Mercedes Urban and Enrique Soto and Bulmaro Cisneros and Ricardo Felix",

note = "Funding Information: Acknowledgments This work was supported by funds from Cona-cyt to RF. We thank Drs. M. E. Mendoza (Cinvestav-IPN, Mexico) and J. C. Gomora (IFC-UNAM, Mexico) for the generous gift of the cell lines and Dr. M. Hernandez (Cinvestav-IPN, Mexico) and D. Mornet (INSERM ERI 25 Muscle et Pathologies, France) for the anti-actin and anti-β-dystroglycan antibodies, respectively. We are also indebted to J. Arikkath for critically reviewing the manuscript. Copyright: Copyright 2010 Elsevier B.V., All rights reserved.",

year = "2009",

month = dec,

day = "1",

doi = "10.1007/s10571-009-9422-2",

language = "English",

volume = "29",

pages = "1265--1273",

journal = "Cellular and Molecular Neurobiology",

issn = "0272-4340",

publisher = "Springer New York",

number = "8",

}

Regulation of Ca<inf>V</inf>3.1 channels by glucocorticoids. / Avila, Traudy; Hernández-Hernández, Oscar; Almanza, Angélica; Bermúdez De León, Mario; Urban, Mercedes; Soto, Enrique; Cisneros, Bulmaro; Felix, Ricardo.

In: Cellular and Molecular Neurobiology, Vol. 29, No. 8, 01.12.2009, p. 1265-1273.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Regulation of CaV3.1 channels by glucocorticoids

AU - Avila, Traudy

AU - Hernández-Hernández, Oscar

AU - Almanza, Angélica

AU - Bermúdez De León, Mario

AU - Urban, Mercedes

AU - Soto, Enrique

AU - Cisneros, Bulmaro

AU - Felix, Ricardo

N1 - Funding Information: Acknowledgments This work was supported by funds from Cona-cyt to RF. We thank Drs. M. E. Mendoza (Cinvestav-IPN, Mexico) and J. C. Gomora (IFC-UNAM, Mexico) for the generous gift of the cell lines and Dr. M. Hernandez (Cinvestav-IPN, Mexico) and D. Mornet (INSERM ERI 25 Muscle et Pathologies, France) for the anti-actin and anti-β-dystroglycan antibodies, respectively. We are also indebted to J. Arikkath for critically reviewing the manuscript. Copyright: Copyright 2010 Elsevier B.V., All rights reserved.

PY - 2009/12/1

Y1 - 2009/12/1

N2 - The activity of low voltage-activated Ca 2+ (Ca V3) channels is tightly coupled to neurotransmitter and hormone secretion. Previous studies have shown that Ca V3 channels are regulated by glucocorticoids (GCs), though the mechanism underlying channel regulation remains unclear. Here, using the pituitary GH 3 cell line as a model, we investigated whether Ca V3 channel expression is under the control of GCs, and if their actions are mediated by transcriptional and/or post-transcriptional mechanisms. RT-PCR and western blot analyses showed that Ca V3.1 but not Ca V3.2 and Ca V3.3 channels is expressed in the GH 3 cells, and patch clamp recordings confirmed that Ca 2+ currents through low voltage-activated channels were decreased after chronic treatment with GCs. Consistent with this, total plasma membrane expression of Ca V3.1 protein as analyzed by cell-surface biotinylation assays and semi-quantitative western blotting was also down-regulated, while quantitative real-time RT-PCR analysis revealed a significant decrease of Ca V3.1 mRNA expression in the treated cells. In contrast, patch-clamp recordings on HEK-293 cells stably expressing recombinant Ca V3.1 channels showed that Ca 2+ currents were not affected by GC treatment. These results suggest that decreased transcription is a likely mechanism to explain the inhibitory actions of GCs on the functional expression of native Ca V3.1 channels.

AB - The activity of low voltage-activated Ca 2+ (Ca V3) channels is tightly coupled to neurotransmitter and hormone secretion. Previous studies have shown that Ca V3 channels are regulated by glucocorticoids (GCs), though the mechanism underlying channel regulation remains unclear. Here, using the pituitary GH 3 cell line as a model, we investigated whether Ca V3 channel expression is under the control of GCs, and if their actions are mediated by transcriptional and/or post-transcriptional mechanisms. RT-PCR and western blot analyses showed that Ca V3.1 but not Ca V3.2 and Ca V3.3 channels is expressed in the GH 3 cells, and patch clamp recordings confirmed that Ca 2+ currents through low voltage-activated channels were decreased after chronic treatment with GCs. Consistent with this, total plasma membrane expression of Ca V3.1 protein as analyzed by cell-surface biotinylation assays and semi-quantitative western blotting was also down-regulated, while quantitative real-time RT-PCR analysis revealed a significant decrease of Ca V3.1 mRNA expression in the treated cells. In contrast, patch-clamp recordings on HEK-293 cells stably expressing recombinant Ca V3.1 channels showed that Ca 2+ currents were not affected by GC treatment. These results suggest that decreased transcription is a likely mechanism to explain the inhibitory actions of GCs on the functional expression of native Ca V3.1 channels.

UR - http://www.scopus.com/inward/record.url?scp=77049109229&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77049109229&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/67f857a6-2d9f-39d8-ac42-367d18ed942b/

U2 - 10.1007/s10571-009-9422-2

DO - 10.1007/s10571-009-9422-2

M3 - Article

VL - 29

SP - 1265

EP - 1273

JO - Cellular and Molecular Neurobiology

JF - Cellular and Molecular Neurobiology

SN - 0272-4340

IS - 8

ER -

Regulation of Ca<inf>V</inf>3.1 channels by glucocorticoids

Abstract

Bibliographical note

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this