Angiotensin-II-Evoked Ca<sup>2+</sup> Entry in Murine Cardiac Fibroblasts Does Not Depend on TRPC Channels
TRPC proteins form cation conducting channels regulated by different stimuli and are regulators of the cellular calcium homeostasis. TRPC are expressed in cardiac cells including cardiac fibroblasts (CFs) and have been implicated in the development of pathological cardiac remodeling including fibros...
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MDPI AG
2020-01-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/2/322 |
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| author | Juan E. Camacho Londoño André Marx Axel E. Kraft Alexander Schürger Christin Richter Alexander Dietrich Peter Lipp Lutz Birnbaumer Marc Freichel |
| author_facet | Juan E. Camacho Londoño André Marx Axel E. Kraft Alexander Schürger Christin Richter Alexander Dietrich Peter Lipp Lutz Birnbaumer Marc Freichel |
| author_sort | Juan E. Camacho Londoño |
| collection | DOAJ |
| description | TRPC proteins form cation conducting channels regulated by different stimuli and are regulators of the cellular calcium homeostasis. TRPC are expressed in cardiac cells including cardiac fibroblasts (CFs) and have been implicated in the development of pathological cardiac remodeling including fibrosis. Using Ca<sup>2+</sup> imaging and several compound TRPC knockout mouse lines we analyzed the involvement of TRPC proteins for the angiotensin II (AngII)-induced changes in Ca<sup>2+</sup> homeostasis in CFs isolated from adult mice. Using qPCR we detected transcripts of all <i>Trpc</i> genes in CFs; <i>Trpc1</i>, <i>Trpc3</i> and <i>Trpc4</i> being the most abundant ones. We show that the AngII-induced Ca<sup>2+</sup> entry but also Ca<sup>2+</sup> release from intracellular stores are critically dependent on the density of CFs in culture and are inversely correlated with the expression of the myofibroblast marker α-smooth muscle actin. Our Ca<sup>2+</sup> measurements depict that the AngII- and thrombin-induced Ca<sup>2+</sup> transients, and the AngII-induced Ca<sup>2+</sup> entry and Ca<sup>2+</sup> release are not affected in CFs isolated from mice lacking all seven TRPC proteins (TRPC-hepta KO) compared to control cells. However, pre-incubation with GSK7975A (10 µM), which sufficiently inhibits CRAC channels in other cells, abolished AngII-induced Ca<sup>2+</sup> entry. Consequently, we conclude the dispensability of the TRPC channels for the acute neurohumoral Ca<sup>2+</sup> signaling evoked by AngII in isolated CFs and suggest the contribution of members of the Orai channel family as molecular constituents responsible for this pathophysiologically important Ca<sup>2+</sup> entry pathway. |
| first_indexed | 2024-03-12T06:27:46Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T06:27:46Z |
| publishDate | 2020-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-29752007b159443c9aeec537c9250d022023-09-03T01:46:16ZengMDPI AGCells2073-44092020-01-019232210.3390/cells9020322cells9020322Angiotensin-II-Evoked Ca<sup>2+</sup> Entry in Murine Cardiac Fibroblasts Does Not Depend on TRPC ChannelsJuan E. Camacho Londoño0André Marx1Axel E. Kraft2Alexander Schürger3Christin Richter4Alexander Dietrich5Peter Lipp6Lutz Birnbaumer7Marc Freichel8Pharmakologisches Institut, Ruprecht-Karls-Universität Heidelberg, INF 366, 69120 Heidelberg, GermanyPharmakologisches Institut, Ruprecht-Karls-Universität Heidelberg, INF 366, 69120 Heidelberg, GermanyPharmakologisches Institut, Ruprecht-Karls-Universität Heidelberg, INF 366, 69120 Heidelberg, GermanyPharmakologisches Institut, Ruprecht-Karls-Universität Heidelberg, INF 366, 69120 Heidelberg, GermanyPharmakologisches Institut, Ruprecht-Karls-Universität Heidelberg, INF 366, 69120 Heidelberg, GermanyWalther-Straub-Institut für Pharmakologie und Toxikologie, Ludwig-Maximilians-Universität, 80336 München, GermanyMedical Faculty, Centre for Molecular Signalling (PZMS), Institute for Molecular Cell Biology and Research Center for Molecular Imaging and Screening, Saarland University, 66421 Homburg/Saar, GermanyLaboratory of Neurobiology, NIEHS, North Carolina, USA and Institute of Biomedical Research (BIOMED), Catholic University of Argentina, Buenos Aires C1107AFF, ArgentinaPharmakologisches Institut, Ruprecht-Karls-Universität Heidelberg, INF 366, 69120 Heidelberg, GermanyTRPC proteins form cation conducting channels regulated by different stimuli and are regulators of the cellular calcium homeostasis. TRPC are expressed in cardiac cells including cardiac fibroblasts (CFs) and have been implicated in the development of pathological cardiac remodeling including fibrosis. Using Ca<sup>2+</sup> imaging and several compound TRPC knockout mouse lines we analyzed the involvement of TRPC proteins for the angiotensin II (AngII)-induced changes in Ca<sup>2+</sup> homeostasis in CFs isolated from adult mice. Using qPCR we detected transcripts of all <i>Trpc</i> genes in CFs; <i>Trpc1</i>, <i>Trpc3</i> and <i>Trpc4</i> being the most abundant ones. We show that the AngII-induced Ca<sup>2+</sup> entry but also Ca<sup>2+</sup> release from intracellular stores are critically dependent on the density of CFs in culture and are inversely correlated with the expression of the myofibroblast marker α-smooth muscle actin. Our Ca<sup>2+</sup> measurements depict that the AngII- and thrombin-induced Ca<sup>2+</sup> transients, and the AngII-induced Ca<sup>2+</sup> entry and Ca<sup>2+</sup> release are not affected in CFs isolated from mice lacking all seven TRPC proteins (TRPC-hepta KO) compared to control cells. However, pre-incubation with GSK7975A (10 µM), which sufficiently inhibits CRAC channels in other cells, abolished AngII-induced Ca<sup>2+</sup> entry. Consequently, we conclude the dispensability of the TRPC channels for the acute neurohumoral Ca<sup>2+</sup> signaling evoked by AngII in isolated CFs and suggest the contribution of members of the Orai channel family as molecular constituents responsible for this pathophysiologically important Ca<sup>2+</sup> entry pathway.https://www.mdpi.com/2073-4409/9/2/322trpc channelscardiac fibroblasts (cfs)ca<sup>2+</sup> release and ca<sup>2+</sup> entryangiotensin ii |
| spellingShingle | Juan E. Camacho Londoño André Marx Axel E. Kraft Alexander Schürger Christin Richter Alexander Dietrich Peter Lipp Lutz Birnbaumer Marc Freichel Angiotensin-II-Evoked Ca<sup>2+</sup> Entry in Murine Cardiac Fibroblasts Does Not Depend on TRPC Channels Cells trpc channels cardiac fibroblasts (cfs) ca<sup>2+</sup> release and ca<sup>2+</sup> entry angiotensin ii |
| title | Angiotensin-II-Evoked Ca<sup>2+</sup> Entry in Murine Cardiac Fibroblasts Does Not Depend on TRPC Channels |
| title_full | Angiotensin-II-Evoked Ca<sup>2+</sup> Entry in Murine Cardiac Fibroblasts Does Not Depend on TRPC Channels |
| title_fullStr | Angiotensin-II-Evoked Ca<sup>2+</sup> Entry in Murine Cardiac Fibroblasts Does Not Depend on TRPC Channels |
| title_full_unstemmed | Angiotensin-II-Evoked Ca<sup>2+</sup> Entry in Murine Cardiac Fibroblasts Does Not Depend on TRPC Channels |
| title_short | Angiotensin-II-Evoked Ca<sup>2+</sup> Entry in Murine Cardiac Fibroblasts Does Not Depend on TRPC Channels |
| title_sort | angiotensin ii evoked ca sup 2 sup entry in murine cardiac fibroblasts does not depend on trpc channels |
| topic | trpc channels cardiac fibroblasts (cfs) ca<sup>2+</sup> release and ca<sup>2+</sup> entry angiotensin ii |
| url | https://www.mdpi.com/2073-4409/9/2/322 |
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