The TRPV4 cation channel mediates stretch-evoked Ca2+ influx and ATP release in primary urothelial cell cultures

Tsutomu Mochizuki, Takaaki Sokabe, Isao Araki, Kayoko Fujishita, Koji Shibasaki, Kunitoshi Uchida, Keiji Naruse, Schuichi Koizumi, Masayuki Takeda, Makoto Tominaga

Research output: Contribution to journalArticle

158 Citations (Scopus)

Abstract

Transient receptor potential channels have recently been implicated in physiological functions in a urogenital system. In this study, we investigated the role of transient receptor potential vanilloid 4 (TRPV4) channels in a stretch sensing mechanism in mouse primary urothelial cell cultures. The selective TRPV4 agonist, 4phorbol 12,13-didecanoate (4PDD) evoked Ca2+ influx in wild-type (WT) urothelial cells, but not in TRPV4-deficient (TRPV4KO) cells. We established a cell-stretch system to investigate stretch-evoked changes in intracellular Ca2+ concentration and ATP release. Stretch stimulation evoked intracellular Ca2+ increases in a stretch speed- and distance-dependent manner in WT and TRPV4KO cells. In TRPV4KO urothelial cells, however, the intracellular Ca2+ increase in response to stretch stimulation was significantly attenuated compared with that in WT cells. Stretch-evoked Ca2+ increases in WT urothelium were partially reduced in the presence of ruthenium red, a broad TRP channel blocker, whereas that in TRPV4KO cells did not show such reduction. Potent ATP release occurred following stretch stimulation or 4PDD administration in WT urothelial cells, which was dramatically suppressed in TRPV4KO cells. Stretch-dependent ATP release was almost completely eliminated in the presence of ruthenium red or in the absence of extracellular Ca2+. These results suggest that TRPV4 senses distension of the bladder urothelium, which is converted to an ATP signal in the micturition reflex pathway during urine storage.

Original languageEnglish
Pages (from-to)21257-21264
Number of pages8
JournalJournal of Biological Chemistry
Volume284
Issue number32
DOIs
Publication statusPublished - Aug 7 2009

Fingerprint

TRPV Cation Channels
Primary Cell Culture
Cell culture
Cations
Adenosine Triphosphate
Ruthenium Red
Transient Receptor Potential Channels
Urothelium
Urogenital System
Urination
Reflex
Urinary Bladder
Urine

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Mochizuki, T., Sokabe, T., Araki, I., Fujishita, K., Shibasaki, K., Uchida, K., ... Tominaga, M. (2009). The TRPV4 cation channel mediates stretch-evoked Ca2+ influx and ATP release in primary urothelial cell cultures. Journal of Biological Chemistry, 284(32), 21257-21264. https://doi.org/10.1074/jbc.M109.020206

The TRPV4 cation channel mediates stretch-evoked Ca2+ influx and ATP release in primary urothelial cell cultures. / Mochizuki, Tsutomu; Sokabe, Takaaki; Araki, Isao; Fujishita, Kayoko; Shibasaki, Koji; Uchida, Kunitoshi; Naruse, Keiji; Koizumi, Schuichi; Takeda, Masayuki; Tominaga, Makoto.

In: Journal of Biological Chemistry, Vol. 284, No. 32, 07.08.2009, p. 21257-21264.

Research output: Contribution to journalArticle

Mochizuki, T, Sokabe, T, Araki, I, Fujishita, K, Shibasaki, K, Uchida, K, Naruse, K, Koizumi, S, Takeda, M & Tominaga, M 2009, 'The TRPV4 cation channel mediates stretch-evoked Ca2+ influx and ATP release in primary urothelial cell cultures', Journal of Biological Chemistry, vol. 284, no. 32, pp. 21257-21264. https://doi.org/10.1074/jbc.M109.020206
Mochizuki, Tsutomu ; Sokabe, Takaaki ; Araki, Isao ; Fujishita, Kayoko ; Shibasaki, Koji ; Uchida, Kunitoshi ; Naruse, Keiji ; Koizumi, Schuichi ; Takeda, Masayuki ; Tominaga, Makoto. / The TRPV4 cation channel mediates stretch-evoked Ca2+ influx and ATP release in primary urothelial cell cultures. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 32. pp. 21257-21264.
@article{da5e3d15e4344965b02c35ed8213a2cc,
title = "The TRPV4 cation channel mediates stretch-evoked Ca2+ influx and ATP release in primary urothelial cell cultures",
abstract = "Transient receptor potential channels have recently been implicated in physiological functions in a urogenital system. In this study, we investigated the role of transient receptor potential vanilloid 4 (TRPV4) channels in a stretch sensing mechanism in mouse primary urothelial cell cultures. The selective TRPV4 agonist, 4phorbol 12,13-didecanoate (4PDD) evoked Ca2+ influx in wild-type (WT) urothelial cells, but not in TRPV4-deficient (TRPV4KO) cells. We established a cell-stretch system to investigate stretch-evoked changes in intracellular Ca2+ concentration and ATP release. Stretch stimulation evoked intracellular Ca2+ increases in a stretch speed- and distance-dependent manner in WT and TRPV4KO cells. In TRPV4KO urothelial cells, however, the intracellular Ca2+ increase in response to stretch stimulation was significantly attenuated compared with that in WT cells. Stretch-evoked Ca2+ increases in WT urothelium were partially reduced in the presence of ruthenium red, a broad TRP channel blocker, whereas that in TRPV4KO cells did not show such reduction. Potent ATP release occurred following stretch stimulation or 4PDD administration in WT urothelial cells, which was dramatically suppressed in TRPV4KO cells. Stretch-dependent ATP release was almost completely eliminated in the presence of ruthenium red or in the absence of extracellular Ca2+. These results suggest that TRPV4 senses distension of the bladder urothelium, which is converted to an ATP signal in the micturition reflex pathway during urine storage.",
author = "Tsutomu Mochizuki and Takaaki Sokabe and Isao Araki and Kayoko Fujishita and Koji Shibasaki and Kunitoshi Uchida and Keiji Naruse and Schuichi Koizumi and Masayuki Takeda and Makoto Tominaga",
year = "2009",
month = "8",
day = "7",
doi = "10.1074/jbc.M109.020206",
language = "English",
volume = "284",
pages = "21257--21264",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "32",

}

TY - JOUR

T1 - The TRPV4 cation channel mediates stretch-evoked Ca2+ influx and ATP release in primary urothelial cell cultures

AU - Mochizuki, Tsutomu

AU - Sokabe, Takaaki

AU - Araki, Isao

AU - Fujishita, Kayoko

AU - Shibasaki, Koji

AU - Uchida, Kunitoshi

AU - Naruse, Keiji

AU - Koizumi, Schuichi

AU - Takeda, Masayuki

AU - Tominaga, Makoto

PY - 2009/8/7

Y1 - 2009/8/7

N2 - Transient receptor potential channels have recently been implicated in physiological functions in a urogenital system. In this study, we investigated the role of transient receptor potential vanilloid 4 (TRPV4) channels in a stretch sensing mechanism in mouse primary urothelial cell cultures. The selective TRPV4 agonist, 4phorbol 12,13-didecanoate (4PDD) evoked Ca2+ influx in wild-type (WT) urothelial cells, but not in TRPV4-deficient (TRPV4KO) cells. We established a cell-stretch system to investigate stretch-evoked changes in intracellular Ca2+ concentration and ATP release. Stretch stimulation evoked intracellular Ca2+ increases in a stretch speed- and distance-dependent manner in WT and TRPV4KO cells. In TRPV4KO urothelial cells, however, the intracellular Ca2+ increase in response to stretch stimulation was significantly attenuated compared with that in WT cells. Stretch-evoked Ca2+ increases in WT urothelium were partially reduced in the presence of ruthenium red, a broad TRP channel blocker, whereas that in TRPV4KO cells did not show such reduction. Potent ATP release occurred following stretch stimulation or 4PDD administration in WT urothelial cells, which was dramatically suppressed in TRPV4KO cells. Stretch-dependent ATP release was almost completely eliminated in the presence of ruthenium red or in the absence of extracellular Ca2+. These results suggest that TRPV4 senses distension of the bladder urothelium, which is converted to an ATP signal in the micturition reflex pathway during urine storage.

AB - Transient receptor potential channels have recently been implicated in physiological functions in a urogenital system. In this study, we investigated the role of transient receptor potential vanilloid 4 (TRPV4) channels in a stretch sensing mechanism in mouse primary urothelial cell cultures. The selective TRPV4 agonist, 4phorbol 12,13-didecanoate (4PDD) evoked Ca2+ influx in wild-type (WT) urothelial cells, but not in TRPV4-deficient (TRPV4KO) cells. We established a cell-stretch system to investigate stretch-evoked changes in intracellular Ca2+ concentration and ATP release. Stretch stimulation evoked intracellular Ca2+ increases in a stretch speed- and distance-dependent manner in WT and TRPV4KO cells. In TRPV4KO urothelial cells, however, the intracellular Ca2+ increase in response to stretch stimulation was significantly attenuated compared with that in WT cells. Stretch-evoked Ca2+ increases in WT urothelium were partially reduced in the presence of ruthenium red, a broad TRP channel blocker, whereas that in TRPV4KO cells did not show such reduction. Potent ATP release occurred following stretch stimulation or 4PDD administration in WT urothelial cells, which was dramatically suppressed in TRPV4KO cells. Stretch-dependent ATP release was almost completely eliminated in the presence of ruthenium red or in the absence of extracellular Ca2+. These results suggest that TRPV4 senses distension of the bladder urothelium, which is converted to an ATP signal in the micturition reflex pathway during urine storage.

UR - http://www.scopus.com/inward/record.url?scp=69249100479&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=69249100479&partnerID=8YFLogxK

U2 - 10.1074/jbc.M109.020206

DO - 10.1074/jbc.M109.020206

M3 - Article

C2 - 19531473

AN - SCOPUS:69249100479

VL - 284

SP - 21257

EP - 21264

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 32

ER -