Cysteine-rich secretory protein 4 is an inhibitor of transient receptor potential M8 with a role in establishing sperm function
Cysteine-rich secretory protein 4 is an inhibitor of transient receptor potential M8 with a role in establishing sperm function
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DOI：
10.1073/pnas.1015935108
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发表时间：
2011-04-26
影响因子：
11.1
通讯作者：
O'Bryan, Moira K.
中科院分区：
文献类型：
--
作者：
Gibbs, Gerard M.;Orta, Gerardo;O'Bryan, Moira K.
富含半胱氨酸的分泌蛋白（CRISPs）是小鼠中的一组四种蛋白质，在雄性生殖道中大量表达，在其他组织中表达程度较低。对爬行动物CRISPs和小鼠CRISP2的分析表明，CRISPs可以通过离子通道调节细胞稳态。除了CRISP2调节ryanodine受体的能力之外，哺乳动物CRISPs功能的体内靶标是未知的。在这项研究中，我们使用电生理学，细胞测定和小鼠模型表征了附睾CRISP4的离子通道调节活性。通过睾丸精子的膜片钳，CRISP4 CRISP结构域显示出抑制瞬时受体电位（TRP）离子通道TRPM8。使用稳定转染的CHO细胞系证实了这些数据。TRPM8是人体内主要的冷受体，但也存在于其他组织中，包括睾丸以及小鼠和人类精子的尾部和头部。使用来自野生型小鼠的精子进行的功能测定表明，TRPM8活化显著减少了获能后经历孕酮诱导的顶体反应的精子数量，并且这种反应被CRISP4的共同添加逆转。因此，Crisp4基因敲除小鼠的精子在接受孕酮诱导的顶体反应方面能力受损。总的来说，这些数据将CRISP4鉴定为TRPM8的内源性调节剂，其在正常精子功能中起作用。
The cysteine-rich secretory proteins (CRISPs) are a group of four proteins in the mouse that are expressed abundantly in the male reproductive tract, and to a lesser extent in other tissues. Analysis of reptile CRISPs and mouse CRISP2 has shown that CRISPs can regulate cellular homeostasis via ion channels. With the exception of the ability of CRISP2 to regulate ryanodine receptors, the in vivo targets of mammalian CRISPs function are unknown. In this study, we have characterized the ion channel regulatory activity of epididymal CRISP4 using electrophysiology, cell assays, and mouse models. Through patch-clamping of testicular sperm, the CRISP4 CRISP domain was shown to inhibit the transient receptor potential (TRP) ion channel TRPM8. These data were confirmed using a stably transfected CHO cell line. TRPM8 is a major cold receptor in the body, but is found in other tissues, including the testis and on the tail and head of mouse and human sperm. Functional assays using sperm from wild-type mice showed that TRPM8 activation significantly reduced the number of sperm undergoing the progesterone-induced acrosome reaction following capacitation, and that this response was reversed by the coaddition of CRISP4. In accordance, sperm from Crisp4 null mice had a compromised ability to undergo to the progesterone-induced acrosome reaction. Collectively, these data identify CRISP4 as an endogenous regulator of TRPM8 with a role in normal sperm function.