ALPHA-2-ADRENERGIC INHIBITION OF CL- TRANSPORT BY OPERCULAR EPITHELIUM IS MEDIATED BY INTRACELLULAR CA2+

被引:36
作者
MARSHALL, WS
BRYSON, SE
GARG, D
机构
[1] Department of Biology, St. Francis Xavier University, Antigonish
关键词
FUNDULUS-HETEROCLITUS; CHLORIDE CELL; IONOMYCIN; THAPSIGARGIN; INTRACELLULAR PH;
D O I
10.1073/pnas.90.12.5504
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We isolated the opercular epithelium of seawater killifish (Fundulus heteroclitus) to study the mediation of catecholamine inhibition of Cl- secretion. The receptors are alpha2-adrenergic, as they have a high affinity for the alpha2-adrenergic agonist clonidine over phenylephrine and clonidine action is blocked by yohimbine. Pertussis toxin and indomethacin did not block the clonidine effect; hence inhibitory guanine nucleotide-binding proteins (G(i) proteins) and prostaglandins (respectively) are not involved. Intracellular pH (pH(i)) of single chloride cells was measured microspectrofluorometrically and resting pH(i) was 7.22 +/- 0.03. However, pH(i) was unaffected by clonidine; hence pH(i) and Na+/H+ exchange are not involved. The lipoxygenase inhibitors nordihydroguaiaretic acid and baicalein and the lipoxygenase products (12S)- and (12R)-12-hydroxyeicosatetraenoic acid stimulated Cl- secretion. Protein kinase C is an unlikely site of action because the diacylglycerol kinase inhibitor R59022 had no effect alone and did not block the clonidine effect. Ionomycin (1 muM) in normal but not low-Ca2+ solutions mimicked the action of clonidine and both inhibitions were reversible by isoproterenol. Thapsigargin, a releaser of intracellular Ca2+, inhibited Cl- secretion and this effect was reduced in low-Ca2+ solutions. Low-Ca2+ solutions also blunted but did not block entirely the clonidine response, indicating that the primary Ca2+ release was from intracellular stores. Whereas alpha1-adrenergic receptors commonly act via the Ca2+/inositol trisphosphate pathway, to our knowledge this is the first report of a Ca2+-mediated alpha2-adrenergic response in a nonmammalian vertebrate.
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页码:5504 / 5508
页数:5
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