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Na/K-ATPase and Regulation of Sperm Function

机译：Na / K-ATPase与精子功能调节

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A standard bull breeding soundness evaluation (BBSE) identifies bulls with semen that is grossly abnormal. Nonetheless, semen samples classified as satisfactory based on these traditional approaches differ in fertility; perhaps there are submicroscopic differences in sperm characteristics affecting fertility. Therefore, a better understanding of molecular regulation of sperm function could promote development of novel, evidence-based approaches to predict male fertility. Recently the α4 isoform of Na/K-ATPase (ATP1A4) has received considerable attention, due to its testis-specific expression in post-meiotic germ cells and mature sperm, in addition to its regulation of sperm motility and capacitation. Using fresh bull sperm, we determined that ATP1A4 resided in specialized microdomains (raft and non-raft) of the sperm plasma membrane and activated specific signaling (caveolin-1, EGFR, Src, ERK1/2) molecules during sperm capacitation. Furthermore, ATP1A4 was the predominant isoform responsible for total Na/K-ATPase activity in capacitated sperm. Despite the widely accepted dogma of transcriptional/translational quiescence, bovine sperm translated ATP1A4 mRNA on mitochondrial or mitochondrial-type ribosomes, increasing their content and activity during capacitation. Proteomic analysis of raft and non-raft fractions revealed a significant interaction between ATP1A4 and plakoglobin, a member of the β-catenin family of proteins involved in cell adhesion, in the equatorial segment of capacitated sperm, suggesting a potential role in sperm-oolemma fusion. In frozen-thawed sperm, ATP1A4 content and activity was greater in high- versus low-fertility bulls. Additionally, ATP1A4-induced increases in ROS, calcium, actin polymerization and tyrosine phosphorylation were also involved in regulating post-thaw sperm function in these bulls. Overall, results demonstrated that ATP1A4 had unique roles in controlling several aspects of sperm physiology, acting through well-established enzyme activity and signaling functions. Consequently, isoforms of Na/K-ATPase are potential biomarkers for male fertility.
机译：标准的公牛繁殖健全性评估（BBSE）可识别出精液严重异常的公牛。尽管如此，根据这些传统方法分类为令人满意的精液样品在生育能力上有所不同。精子特征可能存在亚显微差异，从而影响生育能力。因此，对精子功能的分子调控的更好理解可以促进新的，循证的方法来预测男性的生育能力。最近，由于Na / K-ATPase（ATP1A4）的α4亚型除了调节精子活力和获能外，还因其在减数分裂后的生殖细胞和成熟精子中的睾丸特异性表达而备受关注。使用新鲜的公牛精子，我们确定ATP1A4驻留在精子质膜的特定微区（筏和非筏）中，并在精子获能过程中激活了特定信号（caveolin-1，EGFR，Src，ERK1 / 2）分子。此外，ATP1A4是主要的同工型，负责被精子中的总Na / K-ATPase活性。尽管转录/翻译静止的教条已被广泛接受，但牛精子在线粒体或线粒体型核糖体上翻译了ATP1A4 mRNA，从而在捕获过程中增加了其含量和活性。蛋白质组学分析的筏和非筏部分，揭示了ATP1A4和plakoglobin，参与细胞粘附的β-catenin蛋白家族成员的蛋白，在capacated精子的赤道部分之间的显着相互作用，表明在精子-血肿融合中的潜在作用。在冷冻解冻的精子中，高繁殖力和低繁殖力公牛的ATP1A4含量和活性更高。另外，ATP1A4诱导的ROS，钙，肌动蛋白聚合和酪氨酸磷酸化增加也参与了这些公牛解冻后精子功能的调节。总体而言，结果表明ATP1A4在控制精子生理的多个方面具有独特的作用，它们通过公认的酶活性和信号传导功能起作用。因此，Na / K-ATPase的同工型是男性生育力的潜在生物标记。

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