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首页> 外文期刊>Molecular reproduction and development >Extracellular calcium regulates protein tyrosine phosphorylation through calcium-sensing receptor (CaSR) in stallion sperm
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Extracellular calcium regulates protein tyrosine phosphorylation through calcium-sensing receptor (CaSR) in stallion sperm

机译：细胞外钙通过种马精子中的钙敏感受体（CaSR）调节蛋白质酪氨酸磷酸化

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Protein tyrosine phosphorylation (PY), a hallmark of sperm capacitation, is inhibited by extracellular calcium in stallion sperm. The objective of this study was to determine the presence and influence of the calcium-sensing receptor (CaSR) in this phenomenon. First, the presence of the CaSR was demonstrated in stallion sperm. We then tested its function in these gametes using its inhibitor NPS2143 or its agonist AC34356. Sperm were capacitated for 4hr in modified Whitten's medium with 25mM bicarbonate plus NPS2143 and 2.4mM calcium or AC34356 alone, followed by analysis of PY. Inhibition of CaSR with NPS2143 prevented the calcium-dependent PY inhibition in a dose-dependent manner (5, 10, and 15M) whereas AC34356 (100M) inhibited PY similarly to calcium. Stallion sperm motility and viability significantly decreased in presence of 15M of NPS2143 whereas only sperm motility decreased with 100M of AC34356. CaSR function was also studied in the complete absence of calcium by including 2mM ethylene glycol tetraacetic acid (EGTA); under these conditions, AC34356 again inhibited PY, but this time induced a significant increase in sperm motility. Inhibition of calmodulin by W-7 did not recover the AC34356-mediated PY inhibition. When stallion sperm were incubated under capacitating conditions (calcium, bicarbonate, plus bovine serum albumin) at elevated pH (7.9 or 8.5) AC34356 did not block PY. These results thus elucidate the effect of extracellular conditions on the regulation of CaSR, and point to its modulatory role on stallion sperm PY, motility, and viability. Mol. Reprod. Dev. 83: 236-245, 2016. (c) 2016 Wiley Periodicals, Inc.
机译：蛋白质酪氨酸磷酸化（PY）是精子获能的标志，被种马精子中的细胞外钙抑制。这项研究的目的是确定这种现象中钙敏感受体（CaSR）的存在和影响。首先，在种马精子中证明了CaSR的存在。然后，我们使用其抑制剂NPS2143或激动剂AC34356在这些配子中测试了其功能。将精子在含有25mM碳酸氢盐加NPS2143和2.4mM钙或单独AC34356的改良Whitten培养基中获能4小时，然后进行PY分析。用NPS2143抑制CaSR可以以剂量依赖性方式（5、10和15M）阻止钙依赖性PY抑制，而AC34356（100M）与钙类似地抑制PY。在15M的NPS2143存在下，公马的精子活力和生存能力显着下降，而100M的AC34356仅使精子活力下降。还通过加入2mM乙二醇四乙酸（EGTA）在完全不含钙的条件下研究了CaSR功能。在这些条件下，AC34356再次抑制了PY，但是这次诱导了精子活力的显着提高。 W-7抑制钙调蛋白不能恢复AC34356介导的PY抑制。当将种马精子在增容条件下（钙，碳酸氢盐和牛血清白蛋白）在升高的pH值（7.9或8.5）下孵育时，AC34356不会阻断PY。因此，这些结果阐明了细胞外条件对CaSR调控的影响，并指出了其对种马精子PY，运动性和生存力的调节作用。大声笑责备。开发人员83：236-245，2016。（c）2016 Wiley Periodicals，Inc.。

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