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Sources of calcium in agonist-induced contraction of rat distal colon smooth muscle in vitro.

Abstract：

AIM:To study the origin of calcium necessary for agonist-induced contraction of the distal colon in rats. METHODS:The change in intracellular calcium concentration ([Ca2+]i) evoked by elevating external Ca2+ was detected by fura 2/AM fluorescence. Contractile activity was measured with a force displacement transducer. Tension was continuously monitored and recorded using a Powerlab 4/25T data acquisition system with an ML110 bridge bioelectric physiographic amplifier. RESULTS:Store depletion induced Ca2+ influx had an effect on [Ca2+]i. In nominally Ca2+-free medium, the sarco-endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin (1 mumol/L) increased [Ca2+]i from 68 to 241 nmol/L, and to 458 (P < 0.01) and 1006 nmol/L (P < 0.01), respectively, when 1.5 mmol/L and 3.0 mmol/L extracellular Ca2+ was reintroduced. Furthermore, the change in [Ca2+]i was observed with verapamil (5 micromol/L), La3+ (1 mmol/L) or KCl (40 mmol/L) in the bathing solution. These channels were sensitive to La3+ (P < 0.01), insensitive to verapamil, and voltage independent. In isolated distal colons we found that in normal Krebs solution, contraction induced by acetylcholine (ACh) was partially inhibited by verapamil, and the inhibitory rate was 41% (P < 0.05). On the other hand, in Ca2+-free Krebs solution, ACh induced transient contraction due to Ca2+ release from the intracellular stores. The transient contraction lasted until the Ca2+ store was depleted. Restoration of extracellular Ca2+ in the presence of atropine produced contraction, mainly due to Ca2+ influx. Such contraction was not inhibited by verapamil, but was decreased by La3+ (50 micromol/L) from 0.96 to 0.72 g (P < 0.01). CONCLUSION:The predominant source of activator Ca2+ for the contractile response to agonist is extracellular Ca2+, and intracellular Ca2+ has little role to play in mediating excitation-contraction coupling by agonists in rat distal colon smooth muscle in vitro. The influx of extracellular Ca2+ is mainly mediated through voltage-, receptor- and store-operated Ca2+ channels, which can be used as an alternative to develop new drugs targeted on the dysfunction of digestive tract motility.

journal_name

World J Gastroenterol

journal_title

World journal of gastroenterology

authors

Zhou H,Kong DH,Pan QW,Wang HH

doi

10.3748/wjg.14.1077

subject

Has Abstract

pub_date

2008-02-21 00:00:00

pages

1077-83

issue

eissn

1007-9327

issn

2219-2840

journal_volume

pub_type

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Sources of calcium in agonist-induced contraction of rat distal colon smooth muscle in vitro.