DIRECT EFFECT OF ENDOTHELIN-1 ON THE GRANULOSA-CELLS OF THE PORCINE OVARY

被引:41
作者
KAMADA, S [1 ]
KUBOTA, T [1 ]
HIRATA, Y [1 ]
TAGUCHI, M [1 ]
EGUCHI, S [1 ]
MARUMO, F [1 ]
ASO, T [1 ]
机构
[1] TOKYO MED & DENT UNIV,FAC MED,DEPT INTERNAL MED,BUNKYO KU,TOKYO 113,JAPAN
关键词
D O I
10.1677/joe.0.1340059
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Specific binding sites for endothelin-1 (ET-1), a novel potent vasoconstrictor peptide, as well as the effects of ET-1 on cytosolic free Ca2+ concentration ([Ca2+]i), intracellular total inositol phosphate (IP) generation and steroidogenesis were studied in cultured porcine granulosa cells. Scatchard analysis of a binding study using I-125-labelled ET-1 indicated the presence of a single class of high-affinity binding sites with almost equal affinity for ET-1 and ET-3: the apparent dissociation constant was 0.59 nmol/l and the maximal binding capacity was 1.84 pmol/mg protein. Affinity-labelling of I-125-labelled ET-1 to the membranes using disuccinimidyl tartarate as a cross-linker revealed one major and one minor band with the apparent molecular weights of 32 kDa and 49 kDa respectively. ET-1 dose-dependently (1-100 nmol/l) induced rapid and transient increases in [Ca2+]i in fura-2-labelled cells. ET-1 also dose-dependently stimulated total IPs in cells prelabelled with myo-[H-3]inositol. ET-1 had a slight stimulatory effect on the secretion of progesterone but not of oestradiol from porcine granulosa cells. The present data clearly demonstrate the presence of a non-selective ET receptor (ET(B)) in porcine granulosa cells coupled with phosphoinositide hydrolysis and [Ca2+]i mobilization, and suggest that ET-1 may play some role in the production of progesterone by porcine granulosa cells.
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页码:59 / 66
页数:8
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