Cl- Is Required for HCO3- Entry Necessary for Sperm Capacitation in Guinea Pig: Involvement of a Cl-/HCO3- Exchanger (SLC26A3) and CFTR
Cl- Is Required for HCO3- Entry Necessary for Sperm Capacitation in Guinea Pig: Involvement of a Cl-/HCO3- Exchanger (SLC26A3) and CFTR
复制标题
Cl- 是豚鼠精子获能所需的 HCO3- 进入所必需的：Cl-/HCO3- 交换器 (SLC26A3) 和 CFTR 的参与
DOI：
10.1095/biolreprod.108.068528
复制
发表时间：
2009-01-01
影响因子：
3.6
通讯作者：
Shi, Qi Xian
中科院分区：
文献类型：
--
作者：
Chen, Wen Ying;Xu, Wen Ming;Shi, Qi Xian
我们先前的研究证明囊性纤维化跨膜电导调节因子(CFTR)参与了精子获能所必需的碳酸氢盐的转运；然而，它是直接参与还是间接参与仍不清楚。本研究探讨了在豚鼠精子获能过程中使用氯离子/HCO3-交换器(溶质载体家族26，编号3[SLC26A3])的可能性。在含不同浓度氯离子的培养液中孵育精子，可获得不同比例的获能精子，并呈浓度依赖关系。即使在HCO3-存在的情况下，氯离子的耗尽也会破坏精子的获能能力，反之亦然，这表明这两种阴离子都参与了这一过程。与获能相关的HCO3依赖事件，包括细胞内pH升高、cAMP产生和蛋白质酪氨酸磷酸化，也取决于氯离子浓度。在精子过度激活的运动和精子-卵子融合方面，也观察到了相似的氯离子依赖性和抑制剂敏感性。免疫组织化学染色和Western印迹分析证实CFTR和SLC26A3的表达和定位。综上所述，我们的结果表明，Cl3-是精子获能所必需的HCO3-进入所必需的，这意味着SLC26A3参与了HCO3-的运输，CFTR为Cl2提供了循环途径。
Our previous study demonstrated the involvement of cystic fibrosis transmembrane conductance regulator (CFTR) in transporting bicarbonate that is necessary for sperm capacitation; however, whether its involvement is direct or indirect remains unclear. The present study investigated the possibility of a Cl-/HCO3- exchanger (solute carrier family 26, number 3 [SLC26A3]) operating with CFTR during guinea pig sperm capacitation. Incubating sperm in media with various concentrations of Cl- resulted in varied percentages of capacitated sperm in a concentration-dependent manner. Depletion of Cl-, even in the presence of HCO3-, abolished sperm capacitation and vice versa, indicating the involvement of both anions in the process. Capacitation-associated HCO3--dependent events, including increased intracellular pH, cAMP production, and protein tyrosine phosphorylation, also depend on Cl- concentrations. Similar Cl- dependence and inhibitor sensitivity were observed for sperm-hyperactivated motility and for sperm-egg fusion. The expression and localization of CFTR and SLC26A3 were demonstrated using immunostaining and Western blot analysis. Taken together, our results indicate that Cl- is required for the entry of HCO3- that is necessary for sperm capacitation, implicating the involvement of SLC26A3 in transporting HCO3-, with CFTR providing the recycling pathway for Cl-.