Capacitation-associated alkalization in human sperm is differentially controlled at the subcellular level
Capacitation-associated alkalization in human sperm is differentially controlled at the subcellular level
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DOI：
10.1242/jcs.238816
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发表时间：
2020-01-01
影响因子：
4
通讯作者：
Trevino, Claudia L.
中科院分区：
文献类型：
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作者：
Matamoros-Volante, Arturo;Trevino, Claudia L.
哺乳动物精子获能涉及细胞内pH（pH（i））的精确平衡，但控制该过程的机制尚未完全理解，特别是关于参与pHi调节的蛋白质的时空调节。在这里，我们采用了基于图像的流式细胞术技术结合药理学方法来研究获能过程中亚细胞水平的pHi动态。我们发现，获能诱导后，精子细胞进行细胞内碱化的头部和主片区域。所观察到的局部pHi增加需要HCO 3-的初始摄取，这是由几种与其亚细胞定位一致的蛋白质介导的。Hv 1质子通道（也称为HVCN 1）和cAMP激活的蛋白激酶（蛋白激酶A，PKA）拮抗剂主要在主要部分损害碱化。Na+/HCO 3-协同转运蛋白（NBC）和囊性纤维化跨膜调节因子（CFTR）拮抗剂仅轻度损害碱化，主要在头部。运动性测量表明，抑制碱化的主要部分，防止过度激活的运动的发展。总之，我们的研究结果揭示了pHi的复杂控制机制，并强调了它们在人类精子获能过程中的重要性。
Capacitation in mammalian sperm involves the accurate balance of intracellular pH (pH(i)), but the mechanisms controlling this process are not fully understood, particularly regarding the spatiotemporal regulation of the proteins involved in pHi modulation. Here, we employed an image-based flow cytometry technique combined with pharmacological approaches to study pHi dynamics at the subcellular level during capacitation. We found that, upon capacitation induction, sperm cells undergo intracellular alkalization in the head and principal piece regions. The observed localized pHi increases require the initial uptake of HCO3-, which is mediated by several proteins acting consistently with their subcellular localization. Hv1 proton channel (also known as HVCN1) and cAMP-activated protein kinase (protein kinase A, PKA) antagonists impair alkalization mainly in the principal piece. Na+/HCO3- cotransporter (NBC) and cystic fibrosis transmembrane regulator (CFTR) antagonists impair alkalization only mildly, predominantly in the head. Motility measurements indicate that inhibition of alkalization in the principal piece prevents the development of hyperactivated motility. Altogether, our findings shed light on the complex control mechanisms of pHi and underscore their importance during human sperm capacitation.