Involvement of G Protein-Coupled Receptor 30 (GPR30) in Rapid Action of Estrogen in Primate LHRH Neurons
Involvement of G Protein-Coupled Receptor 30 (GPR30) in Rapid Action of Estrogen in Primate LHRH Neurons
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DOI：
10.1210/me.2008-0299
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发表时间：
2009-03-01
影响因子：
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通讯作者：
Terasawa, Ei
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文献类型：
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作者：
Noel, Sekoni D.;Keen, Kim L.;Terasawa, Ei
在此之前，我们已经报道了17 β -雌二醇（E-2）诱导灵长类动物的LHRH释放激素（LHRH）神经元的放电活动增加。本研究探讨了E-2是否改变LHRH释放以及细胞内钙离子（[Ca2+](i)）振荡模式，以及G蛋白偶联受体30 （GPR30）是否在灵长类LHRH神经元中介导E-2的快速作用。结果表明：1)核膜不透性雌激素E-2（雌激素树状聚合物偶联物）和质膜不透性雌激素E-2- bsa偶联物均能在暴露10 min内刺激LHRH的释放；2)雌激素受体拮抗剂ICI 182780不能阻断E-2诱导的LHRH释放，E-2应用于百日咳毒素处理的细胞不能诱导LHRH释放；3) GPR30 mRNA在嗅觉基板培养中表达，GPR30蛋白在LHRH神经元亚群中表达；4)百日咳毒素处理阻断了e -2诱导的[Ca2+](i)振荡的增加；5)转染小干扰RNA （small interfering RNA, siRNA）敲低GPR30在灵长类LHRH神经元中完全消除了e -2诱导的[Ca2+](i)振荡的变化，而转染对照siRNA则没有；6)在转染GPR30 siRNA的LHRH神经元中，雌激素-树突状聚合物偶联物诱导的[Ca2+](i)振荡的增加也没有发生；7) GPR30激动剂G1导致[Ca2+](i)振荡的变化，与E-2相似。总的来说，E-2诱导灵长类LHRH神经元的快速兴奋作用，并且E-2的这种快速作用似乎部分通过GPR30介导。（分子内分泌学23:349-359,2009）
Previously, we have reported that 17 beta-estradiol (E-2) induces an increase in firing activity of primate LH-releasing hormone (LHRH) neurons. The present study investigates whether E-2 alters LHRH release as well as the pattern of intracellular calcium ([Ca2+](i)) oscillations and whether G protein-coupled receptor 30 (GPR30) plays a role in mediating the rapid E-2 action in primate LHRH neurons. Results are summarized: 1) E-2, the nuclear membrane-impermeable estrogen, estrogendendrimer conjugate, and the plasma membrane-impermeable estrogen, E-2-BSA conjugate, all stimulated LHRH release within 10 min of exposure; 2) whereas the estrogen receptor antagonist, ICI 182,780, did not block the E-2-induced LHRH release, E-2 application to cells treated with pertussis toxin failed to induce LHRH release; 3) GPR30 mRNA was expressed in olfactory placode cultures, and GPR30 protein was expressed in a subset of LHRH neurons; 4) pertussis toxin treatment blocked the E-2-induced increase in [Ca2+](i) oscillations; 5) knockdown of GPR30 in primate LHRH neurons by transfection with small interfering RNA ( siRNA) for GPR30 completely abrogated the E-2-induced changes in [Ca2+](i) oscillations, whereas transfection with control siRNA did not; 6) the estrogen-dendrimer conjugate-induced increase in [Ca2+](i) oscillations also did not occur in LHRH neurons transfected with GPR30 siRNA; and 7) G1, a GPR30 agonist, resulted in changes in [Ca2+](i) oscillations, similar to those observed with E-2. Collectively, E-2 induces a rapid excitatory effect on primate LHRH neurons, and this rapid action of E-2 appears to be mediated, in part, through GPR30. (Molecular Endocrinology 23: 349-359, 2009)