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首页> 外文期刊>Cell and Tissue Research >Content of testis-specific isoform of Na/K-ATPase (ATP1A4) is increased during bovine sperm capacitation through translation in mitochondrial ribosomes
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Content of testis-specific isoform of Na/K-ATPase (ATP1A4) is increased during bovine sperm capacitation through translation in mitochondrial ribosomes

机译：通过在线粒体核糖体中的翻译，在牛精子加固期间增加了Na / K-ATPase（ATP1A4）的睾丸特异性同种型的含量

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摘要

Capacitation comprises a series of structural and functional modifications of sperm that confer fertilizing ability. We previously reported that the testis-specific isoform of Na/K-ATPase (ATP1A4) regulated bovine sperm capacitation through signaling mechanisms involving kinases. During subsequent investigations to elucidate mechanisms by which ATP1A4 regulates sperm capacitation, we observed that ATP1A4 was localised in both raft and non-raft fractions of the sperm plasma membrane and that its total content was increased in both membrane fractions during capacitation. The objective of the present study was to investigate mechanism(s) of capacitation-associated increase in the content of ATP1A4. Despite the widely accepted dogma of transcriptional/translational quiescence, incubation of sperm with either ouabain (specific ligand for ATP1A4) or heparin increased ATP1A4 content in raft and non-raft sperm membrane fractions, total sperm protein extracts (immunoblotting) and fixed sperm (flow cytometry), with a concurrent increase in Na/K-ATPase enzyme activity. This capacitation-associated increase in ATP1A4 content was partially decreased by chloramphenicol (mitochondrial translation inhibitor) but not affected by actinomycin D (transcription inhibitor). To demonstrate de novo ATP1A4 synthesis, we evaluated incorporation of bodipy conjugated lysine in this protein during capacitation. A partial decrease in bodipy-lysine incorporation occurred in ATP1A4 from sperm capacitated in the presence of chloramphenicol. Therefore, increased ATP1A4 content during capacitation was attributed to mitochondrial translation of ATP1A4 mRNA present in ejaculated sperm, rather than gene transcription. To our knowledge, this is the first report demonstrating ATP1A4 synthesis during bovine sperm capacitation.
机译：电容包括一系列结构和功能性修饰，精子赋予施肥能力。我们之前报道了通过涉及激酶的信号机制调节Na / K-ATP酶（ATP1A4）的睾丸特异性同种型。在随后的调查期间，在阐明ATP1A4调节精子加固的机制期间，我们观察到，ATP1A4在精子等离子体膜的筏和非筏级分开，并且其总含量在电容过程中两种膜馏分增加。本研究的目的是调查ATP1A4含量的容量相关增加的机制。尽管众所周度地接受转录/翻译静脉的教条，但用瓦巴因（ATP1A4的特定配体）或肝素在筏子和非筏子精子膜馏分中孵育精子（特定配体）或肝素，总精子蛋白质提取物（免疫印迹）和固定精子（流动Cytometry），并发增加Na / K-ATP酶活性。通过氯霉素（线粒体平版翻译抑制剂）（线粒体平版翻译抑制剂）部分地减少了ATP1a4含量的这种相关的增加，但不受放线菌素D（转录抑制剂）的影响。为了证明De Novo ATP1A4合成，我们在电容过程中评估了在该蛋白质中的Bodipy缀合赖氨酸的掺入。在氯霉素存在下的精子中，在ATP1A4中发生Bodipy-Lysine掺入的部分减少。因此，在电容期间增加ATP1A4含量归因于射精精子的ATP1A4 mRNA的线粒体翻译，而不是基因转录。据我们所知，这是第一份报告在牛精子加工过程中证明ATP1A4合成。

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