Insights into GABA receptor signalling in TM3 Leydig cells

Autores
Doepne, R. F. G,; Geigerseder, C.; Frungieri, Monica Beatriz; Gonzalez Calvar, Silvia I.; Calandra, Ricardo Saul; Raemsch, R.; Fohr K,; Kunz, L.; Mayerhofer, A.
Año de publicación
2005
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Gamma-aminobutyric acid (GABA) is an emerging signalling molecule in endocrine organs, since it is produced by endocrine cells and acts via GABA(A) receptors in a paracrine/autocrine fashion. Testicular Leydig cells are producers and targets for GABA. These cells express GABA(A) receptor subunits and in the murine Leydig cell line TM3 pharmacological activation leads to increased proliferation. The signalling pathway of GABA in these cells is not known in this study. We therefore attempted to elucidate details of GABA(A) signalling in TM3 and adult mouse Leydig cells using several experimental approaches. TM3 cells not only express GABA(A )receptor subunits, but also bind the GABA agonist [(3)H]muscimol with a binding affinity in the range reported for other endocrine cells (K(d) = 2.740 +/- 0.721 nM). However, they exhibit a low B(max) value of 28.08 fmol/mg protein. Typical GABA(A) receptor-associated events, including Cl(-) currents, changes in resting membrane potential, intracellular Ca(2+) or cAMP, were not measurable with the methods employed in TM3 cells, or, as studied in part, in primary mouse Leydig cells. GABA or GABA(A) agonist isoguvacine treatment resulted in increased or decreased levels of several mRNAs, including transcription factors (c-fos, hsf-1, egr-1) and cell cycle-associated genes (Cdk2, cyclin D1). In an attempt to verify the cDNA array results and because egr-1 was recently implied in Leydig cell development, we further studied this factor. RT-PCR and Western blotting confirmed a time-dependent regulation of egr-1 in TM3. In the postnatal testis egr-1 was seen in cytoplasmic and nuclear locations of developing Leydig cells, which bear GABA(A) receptors and correspond well to TM3 cells. Thus, GABA acts via an atypical novel signalling pathway in TM3 cells. Further details of this pathway remain to be elucidated.
Fil: Doepne, R. F. G,. Ludwig Maximilians Universitat; Alemania
Fil: Geigerseder, C.. Ludwig Maximilians Universitat; Alemania
Fil: Frungieri, Monica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Gonzalez Calvar, Silvia I.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Calandra, Ricardo Saul. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Raemsch, R.. Ludwig Maximilians Universitat; Alemania
Fil: Fohr K,. Universitat Ulm; Alemania
Fil: Kunz, L.. Ludwig Maximilians Universitat; Alemania
Fil: Mayerhofer, A.. Ludwig Maximilians Universitat; Alemania
Materia
Leydig Cells
Testis
Aminobutyric Acid
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/31342
Acceder
id CONICETDig_b562b899227e11b1f9d1daf3cf6dc3e3
oai_identifier_str oai:ri.conicet.gov.ar:11336/31342
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Insights into GABA receptor signalling in TM3 Leydig cellsDoepne, R. F. G,Geigerseder, C.Frungieri, Monica BeatrizGonzalez Calvar, Silvia I.Calandra, Ricardo SaulRaemsch, R.Fohr K,Kunz, L.Mayerhofer, A.Leydig CellsTestisAminobutyric Acidhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Gamma-aminobutyric acid (GABA) is an emerging signalling molecule in endocrine organs, since it is produced by endocrine cells and acts via GABA(A) receptors in a paracrine/autocrine fashion. Testicular Leydig cells are producers and targets for GABA. These cells express GABA(A) receptor subunits and in the murine Leydig cell line TM3 pharmacological activation leads to increased proliferation. The signalling pathway of GABA in these cells is not known in this study. We therefore attempted to elucidate details of GABA(A) signalling in TM3 and adult mouse Leydig cells using several experimental approaches. TM3 cells not only express GABA(A )receptor subunits, but also bind the GABA agonist [(3)H]muscimol with a binding affinity in the range reported for other endocrine cells (K(d) = 2.740 +/- 0.721 nM). However, they exhibit a low B(max) value of 28.08 fmol/mg protein. Typical GABA(A) receptor-associated events, including Cl(-) currents, changes in resting membrane potential, intracellular Ca(2+) or cAMP, were not measurable with the methods employed in TM3 cells, or, as studied in part, in primary mouse Leydig cells. GABA or GABA(A) agonist isoguvacine treatment resulted in increased or decreased levels of several mRNAs, including transcription factors (c-fos, hsf-1, egr-1) and cell cycle-associated genes (Cdk2, cyclin D1). In an attempt to verify the cDNA array results and because egr-1 was recently implied in Leydig cell development, we further studied this factor. RT-PCR and Western blotting confirmed a time-dependent regulation of egr-1 in TM3. In the postnatal testis egr-1 was seen in cytoplasmic and nuclear locations of developing Leydig cells, which bear GABA(A) receptors and correspond well to TM3 cells. Thus, GABA acts via an atypical novel signalling pathway in TM3 cells. Further details of this pathway remain to be elucidated.Fil: Doepne, R. F. G,. Ludwig Maximilians Universitat; AlemaniaFil: Geigerseder, C.. Ludwig Maximilians Universitat; AlemaniaFil: Frungieri, Monica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Gonzalez Calvar, Silvia I.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Calandra, Ricardo Saul. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Raemsch, R.. Ludwig Maximilians Universitat; AlemaniaFil: Fohr K,. Universitat Ulm; AlemaniaFil: Kunz, L.. Ludwig Maximilians Universitat; AlemaniaFil: Mayerhofer, A.. Ludwig Maximilians Universitat; AlemaniaKarger2005-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/31342Mayerhofer, A.; Kunz, L.; Fohr K,; Raemsch, R.; Calandra, Ricardo Saul; Gonzalez Calvar, Silvia I.; et al.; Insights into GABA receptor signalling in TM3 Leydig cells; Karger; Neuroendocrinology; 81; 6; 12-2005; 381-3900028-38351423-0194CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.karger.com/Article/Abstract/89556info:eu-repo/semantics/altIdentifier/doi/10.1159/000089556info:eu-repo/semantics/altIdentifier/pmid/https://www.ncbi.nlm.nih.gov/pubmed/16276116info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:42:26Zoai:ri.conicet.gov.ar:11336/31342instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:42:26.309CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Insights into GABA receptor signalling in TM3 Leydig cells
title Insights into GABA receptor signalling in TM3 Leydig cells
spellingShingle Insights into GABA receptor signalling in TM3 Leydig cells
Doepne, R. F. G,
Leydig Cells
Testis
Aminobutyric Acid
title_short Insights into GABA receptor signalling in TM3 Leydig cells
title_full Insights into GABA receptor signalling in TM3 Leydig cells
title_fullStr Insights into GABA receptor signalling in TM3 Leydig cells
title_full_unstemmed Insights into GABA receptor signalling in TM3 Leydig cells
title_sort Insights into GABA receptor signalling in TM3 Leydig cells
dc.creator.none.fl_str_mv Doepne, R. F. G,
Geigerseder, C.
Frungieri, Monica Beatriz
Gonzalez Calvar, Silvia I.
Calandra, Ricardo Saul
Raemsch, R.
Fohr K,
Kunz, L.
Mayerhofer, A.
author Doepne, R. F. G,
author_facet Doepne, R. F. G,
Geigerseder, C.
Frungieri, Monica Beatriz
Gonzalez Calvar, Silvia I.
Calandra, Ricardo Saul
Raemsch, R.
Fohr K,
Kunz, L.
Mayerhofer, A.
author_role author
author2 Geigerseder, C.
Frungieri, Monica Beatriz
Gonzalez Calvar, Silvia I.
Calandra, Ricardo Saul
Raemsch, R.
Fohr K,
Kunz, L.
Mayerhofer, A.
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Leydig Cells
Testis
Aminobutyric Acid
topic Leydig Cells
Testis
Aminobutyric Acid
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Gamma-aminobutyric acid (GABA) is an emerging signalling molecule in endocrine organs, since it is produced by endocrine cells and acts via GABA(A) receptors in a paracrine/autocrine fashion. Testicular Leydig cells are producers and targets for GABA. These cells express GABA(A) receptor subunits and in the murine Leydig cell line TM3 pharmacological activation leads to increased proliferation. The signalling pathway of GABA in these cells is not known in this study. We therefore attempted to elucidate details of GABA(A) signalling in TM3 and adult mouse Leydig cells using several experimental approaches. TM3 cells not only express GABA(A )receptor subunits, but also bind the GABA agonist [(3)H]muscimol with a binding affinity in the range reported for other endocrine cells (K(d) = 2.740 +/- 0.721 nM). However, they exhibit a low B(max) value of 28.08 fmol/mg protein. Typical GABA(A) receptor-associated events, including Cl(-) currents, changes in resting membrane potential, intracellular Ca(2+) or cAMP, were not measurable with the methods employed in TM3 cells, or, as studied in part, in primary mouse Leydig cells. GABA or GABA(A) agonist isoguvacine treatment resulted in increased or decreased levels of several mRNAs, including transcription factors (c-fos, hsf-1, egr-1) and cell cycle-associated genes (Cdk2, cyclin D1). In an attempt to verify the cDNA array results and because egr-1 was recently implied in Leydig cell development, we further studied this factor. RT-PCR and Western blotting confirmed a time-dependent regulation of egr-1 in TM3. In the postnatal testis egr-1 was seen in cytoplasmic and nuclear locations of developing Leydig cells, which bear GABA(A) receptors and correspond well to TM3 cells. Thus, GABA acts via an atypical novel signalling pathway in TM3 cells. Further details of this pathway remain to be elucidated.
Fil: Doepne, R. F. G,. Ludwig Maximilians Universitat; Alemania
Fil: Geigerseder, C.. Ludwig Maximilians Universitat; Alemania
Fil: Frungieri, Monica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Gonzalez Calvar, Silvia I.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Calandra, Ricardo Saul. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Raemsch, R.. Ludwig Maximilians Universitat; Alemania
Fil: Fohr K,. Universitat Ulm; Alemania
Fil: Kunz, L.. Ludwig Maximilians Universitat; Alemania
Fil: Mayerhofer, A.. Ludwig Maximilians Universitat; Alemania
description Gamma-aminobutyric acid (GABA) is an emerging signalling molecule in endocrine organs, since it is produced by endocrine cells and acts via GABA(A) receptors in a paracrine/autocrine fashion. Testicular Leydig cells are producers and targets for GABA. These cells express GABA(A) receptor subunits and in the murine Leydig cell line TM3 pharmacological activation leads to increased proliferation. The signalling pathway of GABA in these cells is not known in this study. We therefore attempted to elucidate details of GABA(A) signalling in TM3 and adult mouse Leydig cells using several experimental approaches. TM3 cells not only express GABA(A )receptor subunits, but also bind the GABA agonist [(3)H]muscimol with a binding affinity in the range reported for other endocrine cells (K(d) = 2.740 +/- 0.721 nM). However, they exhibit a low B(max) value of 28.08 fmol/mg protein. Typical GABA(A) receptor-associated events, including Cl(-) currents, changes in resting membrane potential, intracellular Ca(2+) or cAMP, were not measurable with the methods employed in TM3 cells, or, as studied in part, in primary mouse Leydig cells. GABA or GABA(A) agonist isoguvacine treatment resulted in increased or decreased levels of several mRNAs, including transcription factors (c-fos, hsf-1, egr-1) and cell cycle-associated genes (Cdk2, cyclin D1). In an attempt to verify the cDNA array results and because egr-1 was recently implied in Leydig cell development, we further studied this factor. RT-PCR and Western blotting confirmed a time-dependent regulation of egr-1 in TM3. In the postnatal testis egr-1 was seen in cytoplasmic and nuclear locations of developing Leydig cells, which bear GABA(A) receptors and correspond well to TM3 cells. Thus, GABA acts via an atypical novel signalling pathway in TM3 cells. Further details of this pathway remain to be elucidated.
publishDate 2005
dc.date.none.fl_str_mv 2005-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/31342
Mayerhofer, A.; Kunz, L.; Fohr K,; Raemsch, R.; Calandra, Ricardo Saul; Gonzalez Calvar, Silvia I.; et al.; Insights into GABA receptor signalling in TM3 Leydig cells; Karger; Neuroendocrinology; 81; 6; 12-2005; 381-390
0028-3835
1423-0194
CONICET Digital
CONICET
url http://hdl.handle.net/11336/31342
identifier_str_mv Mayerhofer, A.; Kunz, L.; Fohr K,; Raemsch, R.; Calandra, Ricardo Saul; Gonzalez Calvar, Silvia I.; et al.; Insights into GABA receptor signalling in TM3 Leydig cells; Karger; Neuroendocrinology; 81; 6; 12-2005; 381-390
0028-3835
1423-0194
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.karger.com/Article/Abstract/89556
info:eu-repo/semantics/altIdentifier/doi/10.1159/000089556
info:eu-repo/semantics/altIdentifier/pmid/https://www.ncbi.nlm.nih.gov/pubmed/16276116
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Karger
publisher.none.fl_str_mv Karger
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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