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ATP induces contraction mediated by the P2Y(2) receptor in rat intestinal subepithelial myofibroblasts.

Abstract：

:Intestinal subepithelial myofibroblasts (IMFs) exist just under the epithelial membrane directly facing the mucosal microvascular capillary surface distributed in the lamina propria. In the gastrointestinal tract, ATP is released from epithelial and endothelial cells in response to mechanical stimuli. Although it has been reported that mechanical stimuli evoke synchronized Ca(2+) waves in cultured IMFs, the contractile responses by ATP stimulation have not been examined. The aim of this study was to clarify the mechanism of the contraction of IMFs in response to ATP. ATP (1-30μM) induced contraction in a concentration-dependent manner. These contractions were inhibited by LaCl(3) (100-300μM) and by Ca(2+)-free solution (0.5mM EGTA). Fura-2/Ca(2+) signals indicated that ATP (1-10μM) elicited transient increases in intracellular Ca(2+) concentration ([Ca(2+)](i)). In addition, αβ-methylene-ATP (10, 30 and 300μM), a broad spectrum P2X agonist at a concentration higher than 100μM, induced neither contraction nor [Ca(2+)](i) rise. UTP (1-30μM), a selective P2Y(2) and P2Y(4) agonist in rodent, induced concentration-dependent contractions and [Ca(2+)](i) increases, whereas ADP and UDP (10μM) did not induce contractions. Pretreatment with suramin (30-100μM), a relatively selective P2Y(2) antagonist, strongly inhibited ATP- and UTP-induced contractions and [Ca(2+)](i) increases. However, pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS: 10-30μM), a receptor antagonist for several P2X and P2Y but less effective to P2Y(2) receptor, failed to inhibit ATP- and UTP-induced contractions and [Ca(2+)](i) increases. By RT-PCR, mRNA expressions of the P2Y(1) and P2Y(2) receptors, but not P2Y(4) or P2Y(6), were detected in IMFs. These results suggest that ATP induces [Ca(2+)](i)-dependent contraction in IMFs, which is mediated through the P2Y(2) receptor.

journal_name

Eur J Pharmacol

journal_title

European journal of pharmacology

authors

Nakamura T,Iwanaga K,Murata T,Hori M,Ozaki H

doi

10.1016/j.ejphar.2011.01.047

subject

Has Abstract

pub_date

2011-04-25 00:00:00

pages

152-8

issue

1-3

eissn

0014-2999

issn

1879-0712

pii

S0014-2999(11)00105-1

journal_volume

657

pub_type

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ATP induces contraction mediated by the P2Y(2) receptor in rat intestinal subepithelial myofibroblasts.