Ouabain Increases Gap Junctional Communication in Epithelial Cells

Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apica...
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Gespeichert in:

Autor*in:	Arturo Ponce [verfasserIn] Isabel Larre [verfasserIn] Aida Castillo [verfasserIn] Refugio García-Villegas [verfasserIn] Adrián Romero [verfasserIn] Catalina Flores-Maldonado [verfasserIn] Jacqueline Martinez-Rendón [verfasserIn] Rubén Gerardo Contreras [verfasserIn] Marcelino Cereijido [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2014

Schlagwörter:	Sodium pump Gap junctions Connexin Neurobiotin MDCK

Übergeordnetes Werk:	In: Cellular Physiology and Biochemistry - Cell Physiol Biochem Press GmbH & Co KG, 2002, 34(2014), 6, Seite 2081-2090
Übergeordnetes Werk:	volume:34 ; year:2014 ; number:6 ; pages:2081-2090

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc Journal toc

DOI / URN:	10.1159/000366403

Katalog-ID:	DOAJ016108051

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520			\|a Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. Conclusion: Ouabain 10 nM increases GJC in MDCK cells.
650		4	\|a Sodium pump
650		4	\|a Gap junctions
650		4	\|a Connexin
650		4	\|a Neurobiotin
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653		0	\|a Physiology
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700	0		\|a Adrián Romero \|e verfasserin \|4 aut
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700	0		\|a Jacqueline Martinez-Rendón \|e verfasserin \|4 aut
700	0		\|a Rubén Gerardo Contreras \|e verfasserin \|4 aut
700	0		\|a Marcelino Cereijido \|e verfasserin \|4 aut
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allfields	10.1159/000366403 doi (DE-627)DOAJ016108051 (DE-599)DOAJ76bc70779a4740d7b9fbda774d3380de DE-627 ger DE-627 rakwb eng QP1-981 QD415-436 Arturo Ponce verfasserin aut Ouabain Increases Gap Junctional Communication in Epithelial Cells 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. Conclusion: Ouabain 10 nM increases GJC in MDCK cells. Sodium pump Gap junctions Connexin Neurobiotin MDCK Physiology Biochemistry Isabel Larre verfasserin aut Aida Castillo verfasserin aut Refugio García-Villegas verfasserin aut Adrián Romero verfasserin aut Catalina Flores-Maldonado verfasserin aut Jacqueline Martinez-Rendón verfasserin aut Rubén Gerardo Contreras verfasserin aut Marcelino Cereijido verfasserin aut In Cellular Physiology and Biochemistry Cell Physiol Biochem Press GmbH & Co KG, 2002 34(2014), 6, Seite 2081-2090 (DE-627)300189702 (DE-600)1482056-0 14219778 nnns volume:34 year:2014 number:6 pages:2081-2090 https://doi.org/10.1159/000366403 kostenfrei https://doaj.org/article/76bc70779a4740d7b9fbda774d3380de kostenfrei http://www.karger.com/Article/FullText/366403 kostenfrei https://doaj.org/toc/1015-8987 Journal toc kostenfrei https://doaj.org/toc/1421-9778 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_374 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2018 GBV_ILN_2153 GBV_ILN_2885 GBV_ILN_2886 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 34 2014 6 2081-2090
spelling	10.1159/000366403 doi (DE-627)DOAJ016108051 (DE-599)DOAJ76bc70779a4740d7b9fbda774d3380de DE-627 ger DE-627 rakwb eng QP1-981 QD415-436 Arturo Ponce verfasserin aut Ouabain Increases Gap Junctional Communication in Epithelial Cells 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. Conclusion: Ouabain 10 nM increases GJC in MDCK cells. Sodium pump Gap junctions Connexin Neurobiotin MDCK Physiology Biochemistry Isabel Larre verfasserin aut Aida Castillo verfasserin aut Refugio García-Villegas verfasserin aut Adrián Romero verfasserin aut Catalina Flores-Maldonado verfasserin aut Jacqueline Martinez-Rendón verfasserin aut Rubén Gerardo Contreras verfasserin aut Marcelino Cereijido verfasserin aut In Cellular Physiology and Biochemistry Cell Physiol Biochem Press GmbH & Co KG, 2002 34(2014), 6, Seite 2081-2090 (DE-627)300189702 (DE-600)1482056-0 14219778 nnns volume:34 year:2014 number:6 pages:2081-2090 https://doi.org/10.1159/000366403 kostenfrei https://doaj.org/article/76bc70779a4740d7b9fbda774d3380de kostenfrei http://www.karger.com/Article/FullText/366403 kostenfrei https://doaj.org/toc/1015-8987 Journal toc kostenfrei https://doaj.org/toc/1421-9778 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_374 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2018 GBV_ILN_2153 GBV_ILN_2885 GBV_ILN_2886 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 34 2014 6 2081-2090
allfields_unstemmed	10.1159/000366403 doi (DE-627)DOAJ016108051 (DE-599)DOAJ76bc70779a4740d7b9fbda774d3380de DE-627 ger DE-627 rakwb eng QP1-981 QD415-436 Arturo Ponce verfasserin aut Ouabain Increases Gap Junctional Communication in Epithelial Cells 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. Conclusion: Ouabain 10 nM increases GJC in MDCK cells. Sodium pump Gap junctions Connexin Neurobiotin MDCK Physiology Biochemistry Isabel Larre verfasserin aut Aida Castillo verfasserin aut Refugio García-Villegas verfasserin aut Adrián Romero verfasserin aut Catalina Flores-Maldonado verfasserin aut Jacqueline Martinez-Rendón verfasserin aut Rubén Gerardo Contreras verfasserin aut Marcelino Cereijido verfasserin aut In Cellular Physiology and Biochemistry Cell Physiol Biochem Press GmbH & Co KG, 2002 34(2014), 6, Seite 2081-2090 (DE-627)300189702 (DE-600)1482056-0 14219778 nnns volume:34 year:2014 number:6 pages:2081-2090 https://doi.org/10.1159/000366403 kostenfrei https://doaj.org/article/76bc70779a4740d7b9fbda774d3380de kostenfrei http://www.karger.com/Article/FullText/366403 kostenfrei https://doaj.org/toc/1015-8987 Journal toc kostenfrei https://doaj.org/toc/1421-9778 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_374 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2018 GBV_ILN_2153 GBV_ILN_2885 GBV_ILN_2886 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 34 2014 6 2081-2090
allfieldsGer	10.1159/000366403 doi (DE-627)DOAJ016108051 (DE-599)DOAJ76bc70779a4740d7b9fbda774d3380de DE-627 ger DE-627 rakwb eng QP1-981 QD415-436 Arturo Ponce verfasserin aut Ouabain Increases Gap Junctional Communication in Epithelial Cells 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. Conclusion: Ouabain 10 nM increases GJC in MDCK cells. Sodium pump Gap junctions Connexin Neurobiotin MDCK Physiology Biochemistry Isabel Larre verfasserin aut Aida Castillo verfasserin aut Refugio García-Villegas verfasserin aut Adrián Romero verfasserin aut Catalina Flores-Maldonado verfasserin aut Jacqueline Martinez-Rendón verfasserin aut Rubén Gerardo Contreras verfasserin aut Marcelino Cereijido verfasserin aut In Cellular Physiology and Biochemistry Cell Physiol Biochem Press GmbH & Co KG, 2002 34(2014), 6, Seite 2081-2090 (DE-627)300189702 (DE-600)1482056-0 14219778 nnns volume:34 year:2014 number:6 pages:2081-2090 https://doi.org/10.1159/000366403 kostenfrei https://doaj.org/article/76bc70779a4740d7b9fbda774d3380de kostenfrei http://www.karger.com/Article/FullText/366403 kostenfrei https://doaj.org/toc/1015-8987 Journal toc kostenfrei https://doaj.org/toc/1421-9778 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_374 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2018 GBV_ILN_2153 GBV_ILN_2885 GBV_ILN_2886 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 34 2014 6 2081-2090
allfieldsSound	10.1159/000366403 doi (DE-627)DOAJ016108051 (DE-599)DOAJ76bc70779a4740d7b9fbda774d3380de DE-627 ger DE-627 rakwb eng QP1-981 QD415-436 Arturo Ponce verfasserin aut Ouabain Increases Gap Junctional Communication in Epithelial Cells 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. Conclusion: Ouabain 10 nM increases GJC in MDCK cells. Sodium pump Gap junctions Connexin Neurobiotin MDCK Physiology Biochemistry Isabel Larre verfasserin aut Aida Castillo verfasserin aut Refugio García-Villegas verfasserin aut Adrián Romero verfasserin aut Catalina Flores-Maldonado verfasserin aut Jacqueline Martinez-Rendón verfasserin aut Rubén Gerardo Contreras verfasserin aut Marcelino Cereijido verfasserin aut In Cellular Physiology and Biochemistry Cell Physiol Biochem Press GmbH & Co KG, 2002 34(2014), 6, Seite 2081-2090 (DE-627)300189702 (DE-600)1482056-0 14219778 nnns volume:34 year:2014 number:6 pages:2081-2090 https://doi.org/10.1159/000366403 kostenfrei https://doaj.org/article/76bc70779a4740d7b9fbda774d3380de kostenfrei http://www.karger.com/Article/FullText/366403 kostenfrei https://doaj.org/toc/1015-8987 Journal toc kostenfrei https://doaj.org/toc/1421-9778 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_374 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2018 GBV_ILN_2153 GBV_ILN_2885 GBV_ILN_2886 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 34 2014 6 2081-2090
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authorswithroles_txt_mv	Arturo Ponce @@aut@@ Isabel Larre @@aut@@ Aida Castillo @@aut@@ Refugio García-Villegas @@aut@@ Adrián Romero @@aut@@ Catalina Flores-Maldonado @@aut@@ Jacqueline Martinez-Rendón @@aut@@ Rubén Gerardo Contreras @@aut@@ Marcelino Cereijido @@aut@@
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callnumber-first	Q - Science
author	Arturo Ponce
spellingShingle	Arturo Ponce misc QP1-981 misc QD415-436 misc Sodium pump misc Gap junctions misc Connexin misc Neurobiotin misc MDCK misc Physiology misc Biochemistry Ouabain Increases Gap Junctional Communication in Epithelial Cells
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topic_title	QP1-981 QD415-436 Ouabain Increases Gap Junctional Communication in Epithelial Cells Sodium pump Gap junctions Connexin Neurobiotin MDCK
topic	misc QP1-981 misc QD415-436 misc Sodium pump misc Gap junctions misc Connexin misc Neurobiotin misc MDCK misc Physiology misc Biochemistry
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title	Ouabain Increases Gap Junctional Communication in Epithelial Cells
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title_full	Ouabain Increases Gap Junctional Communication in Epithelial Cells
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author_browse	Arturo Ponce Isabel Larre Aida Castillo Refugio García-Villegas Adrián Romero Catalina Flores-Maldonado Jacqueline Martinez-Rendón Rubén Gerardo Contreras Marcelino Cereijido
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title_sort	ouabain increases gap junctional communication in epithelial cells
callnumber	QP1-981
title_auth	Ouabain Increases Gap Junctional Communication in Epithelial Cells
abstract	Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. Conclusion: Ouabain 10 nM increases GJC in MDCK cells.
abstractGer	Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. Conclusion: Ouabain 10 nM increases GJC in MDCK cells.
abstract_unstemmed	Background/Aims: The finding that endogenous ouabain acts as a hormone prompted efforts to elucidate its physiological function. In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. Conclusion: Ouabain 10 nM increases GJC in MDCK cells.
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title_short	Ouabain Increases Gap Junctional Communication in Epithelial Cells
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In previous studies, we have shown that 10 nM ouabain (i.e., a concentration within the physiological range) modulates cell-cell contacts such as tight junctions and apical/basolateral polarity. In this study, we examined whether 10 nM ouabain affects another important cell-cell feature: gap junction communication (GJC). Methods: We employed two different approaches: 1) analysis of the cell-to-cell diffusion of neurobiotin injected into a particular MDCK cell (epithelial cells from dog kidneys) in a confluent monolayer by counting the number of neighboring cells reached by the probe and 2) measurement of the electrical capacitance. Results: We found that 10 nM ouabain increase GJC by 475% within 1 hour. The Na+-K+-ATPase acts as a receptor of ouabain. In previous works we have shown that ouabain activates c-Src and ERK1/2 in 1 hour; in the present study we show that the inhibition of these proteins block the effect of ouabain on GJC. This increase in GJC does not require synthesis of new protein components, because the inhibitors cycloheximide and actinomycin D did not affect this phenomenon. Using silencing assays we also demonstrate that this ouabain-induced enhancement of GJC involves connexins 32 and 43. 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Ouabain Increases Gap Junctional Communication in Epithelial Cells

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