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Dynamics of cattle sperm sncRNAs during maturation, from testis to ejaculated sperm

机译：成熟过程中牛精子Sncrnas的动态，从睾丸到射精精子

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During epididymal transit, spermatozoa go through several functional maturation steps, resulting from interactions with epididymal secretomes specific to each region. In particular, the sperm membrane is under constant remodeling, with sequential attachment and shedding of various molecules provided by the epididymal lumen fluid and epididymosomes, which also deliver sncRNA cargo to sperm. As a result, the payload of sperm sncRNAs changes during the transit from the epididymis caput to the cauda. This work was designed to study the dynamics of cattle sperm sncRNAs from spermatogenesis to final maturation. Comprehensive catalogues of sperm sncRNAs were obtained from testicular parenchyma, epididymal caput, corpus and cauda, as well as ejaculated semen from three Holstein bulls. The primary cattle sncRNA sperm content is markedly remodeled as sperm mature along the epididymis. Expression of piRNAs, which are abundant in testis parenchyma, decreases dramatically at epididymis. Conversely, sperm progressively acquires miRNAs, rsRNAs, and tsRNAs along epididymis, with regional specificities. For instance, miRNAs and tsRNAs are enriched in epididymis cauda and ejaculated sperm, while rsRNA expression peaks at epididymis corpus. In addition, epididymis corpus contains mainly 20?nt long piRNAs, instead of 30?nt in all other locations. Beyond the bulk differences in abundance of sncRNAs classes, K-means clustering was performed to study their spatiotemporal expression profile, highlighting differences in specific sncRNAs and providing insights into their putative biological role at each maturation stage. For instance, Gene Ontology analyses using miRNA targets highlighted enriched processes such as cell cycle regulation, response to stress and ubiquitination processes in testicular parenchyma, protein metabolism in epididymal sperm, and embryonic morphogenesis in ejaculated sperm. Our findings confirm that the sperm sncRNAome does not simply reflect a legacy of spermatogenesis. Instead, sperm sncRNA expression shows a remarkable level of plasticity resulting probably from the combination of multiple factors such as loss of the cytoplasmic droplet, interaction with epididymosomes, and more surprisingly, the putative in situ production and/or modification of sncRNAs by sperm. Given the suggested role of sncRNA in epigenetic trans-generational inheritance, our detailed spatiotemporal analysis may pave the way for a study of sperm sncRNAs role in embryo development.
机译：在附睾转运期间，精子通过几种功能成熟步骤，由每个区域特异的与附睾泌酯的相互作用相互作用。特别地，精子膜处于恒定的重塑，具有由附睾内流体和附睾内提供的各种分子的顺序附着和脱落，这也将SNCRNA货物递送至精子。结果，精子SnCRNA的有效载荷在从附睾调节到Cauda的转发过程中发生变化。这项工作旨在研究从精子发生到最终成熟的牛精子Sncrnas的动态。从睾丸实质，附睾成本，菌根和尾部获得综合性精子SnCRNA，以及来自三个Holstein公牛的射精精液。主要牛SncraNA精子含量明显地重新耦合作为沿附睾成熟的精子。 PiRNA的表达在睾丸实质中丰富，在附睾中显着降低。相反，精子逐渐逐渐获取沿着附睾，具有区域特异性的麦芽糖，rsrNA和TSRNA。例如，MiRNA和TSRNA富集在附睾和射出的精子中，而逆相邻语料的RSRNA表达峰。此外，附睾毒品PIRNA主要包含20？NT长PIRNA，而不是在所有其他位置中的30个？NT。除了批量差异的SncrNA类的差异之外，K-Means聚类是进行的，以研究它们的时空表达谱，突出显示特定SnCRNA的差异，并在每个成熟阶段提供其推定的生物学作用的见解。例如，使用miRNA靶标突出显示富集的方法，例如细胞周期调节，对睾丸精子蛋白质代谢的应激和泛素化过程以及射精精子中的胚胎形态发生的响应富集的方法。我们的研究结果证实，精子SnCRNA组并不能简单地反映精子发生的遗产。相反，精子SnCRNA表达显示出显着的塑性水平，可能是由于多种因素的组合，例如细胞质液滴的损失，与附睾的相互作用，更令人惊讶地，通过精子推出原位生产和/或改性SnCRNA的抑制。鉴于SNCRNA在表观遗传经历生遗传中的建议作用，我们的详细时尚分析可能会铺平胚胎发育中精子怠速的作用。

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