Gα12/13-mediated up-regulation of TRPC6 negatively regulates endothelin-1-induced cardiac myofibroblast formation and collagen synthesis through nuclear factor of activated T cells activation

Motohiro Nishida, Naoya Onohara, Yoji Sato, Reiko Suda, Mariko Ogushi, Shihori Tanabe, Ryuji Inoue, Yasuo Mori, Hitoshi Kurose

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Abstract

Sustained elevation of [Ca2+]i has been implicated in many cellular events. We previously reported that α subunits of G 12 family G proteins (Gα12/13) participate in sustained Ca2+ influx required for the activation of nuclear factor of activated T cells (NFAT), a Ca2+-responsive transcriptional factor, in rat neonatal cardiac fibroblasts. Here, we demonstrate that Gα12/13-mediated up-regulation of canonical transient receptor potential 6 (TRPC6) channels participates in sustained Ca2+ influx and NFAT activation by endothelin (ET)-1 treatment. Expression of constitutively active Gα12 or Gα13 increased the expression of TRPC6 proteins and basal Ca2+ influx activity. The treatment with ET-1 increased TRPC6 protein levels through Gα12/13, reactive oxygen species, and c-Jun N-terminal kinase (JNK)-dependent pathways. NFAT is activated by sustained increase in [Ca2+]i through up-regulated TRPC6. A Gα12/13-inhibitory polypeptide derived from the regulator of the G-protein signaling domain of p115-Rho guanine nucleotide exchange factor and a JNK inhibitor, SP600125, suppressed the ET-1-induced increase in expression of marker proteins of myofibroblast formation through a Gα12/13-reactive oxygen species-JNK pathway. The ET-1-induced myofibroblast formation was suppressed by overexpression of TRPC6 and CA NFAT, whereas it was enhanced by TRPC6 small interfering RNAs and cyclosporine A. These results suggest two opposite roles of Gα12/13 in cardiac fibroblasts. First, Gα12/13 mediate ET-1-induced myofibroblast formation. Second, Gα12/13 mediate TRPC6 up-regulation and NFAT activation that negatively regulates ET-1-induced myofibroblast formation. Furthermore, TRPC6 mediates hypertrophic responses in cardiac myocytes but suppresses fibrotic responses in cardiac fibroblasts. Thus, TRPC6 mediates opposite responses in cardiac myocytes and fibroblasts.

Original languageEnglish
Pages (from-to)23117-23128
Number of pages12
JournalJournal of Biological Chemistry
Volume282
Issue number32
DOIs
Publication statusPublished - Aug 10 2007

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NFATC Transcription Factors
Myofibroblasts
Endothelin-1
Up-Regulation
Collagen
Chemical activation
Fibroblasts
Cardiac Myocytes
G12-G13 GTP-Binding Protein alpha Subunits
Reactive Oxygen Species
Phosphotransferases
Rho Guanine Nucleotide Exchange Factors
GTP-Binding Protein Regulators
Proteins
JNK Mitogen-Activated Protein Kinases
GTP-Binding Proteins
Small Interfering RNA
Cyclosporine
Rats
Peptides

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Gα12/13-mediated up-regulation of TRPC6 negatively regulates endothelin-1-induced cardiac myofibroblast formation and collagen synthesis through nuclear factor of activated T cells activation. / Nishida, Motohiro; Onohara, Naoya; Sato, Yoji; Suda, Reiko; Ogushi, Mariko; Tanabe, Shihori; Inoue, Ryuji; Mori, Yasuo; Kurose, Hitoshi.

In: Journal of Biological Chemistry, Vol. 282, No. 32, 10.08.2007, p. 23117-23128.

Research output: Contribution to journalArticle

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AU - Nishida, Motohiro

AU - Onohara, Naoya

AU - Sato, Yoji

AU - Suda, Reiko

AU - Ogushi, Mariko

AU - Tanabe, Shihori

AU - Inoue, Ryuji

AU - Mori, Yasuo

AU - Kurose, Hitoshi

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