Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland.
Institute of Ichthyobiology and Aquaculture, Polish Academy of Sciences, Gołysz, Poland.
J Proteomics. 2019 Jun 30;202:103369. doi: 10.1016/j.jprot.2019.04.019. Epub 2019 Apr 25.
Hormonal stimulation in common carp is a routine practice to enhance sperm production and control gamete maturation. This study aimed to compare the proteome of carp seminal plasma between control and Ovopel-induced males using two-dimensional differential in-gel electrophoresis. Ovopel induction increased sperm volume, total sperm count, seminal plasma osmolality, and pH and decreased seminal plasma protein concentration. In total, 36 spots were identified (23 up- and 13 downregulated), corresponding to 23 proteins differentially abundant in seminal plasma after Ovopel induction (p < .05; fold change 1.2). The majority of proteins were associated with the immune and stress responses including the transport protein (hephaestin), antiproteases (fetuin, α2-macroglobulin, TIMP2), complement components (C3, complement factor B/C2A), regulator of the coagulation cascade (plasminogen), modulators of the innate immune response, such as intelectin, ApoA and ApoE, and the cathepsin/cystatin system, and stress response (enolase1). In addition, hormonal stimulation seems to be related to the proteins involved in lipid metabolism, signal transduction, and tissue remodeling. Our results suggest that hormonal stimulation is not just concomitant with the hydration of testis but also induces the synthesis and secretion of seminal plasma proteins involved in sperm maturation and protection against stress induced by administration of the exogenous hormone. SIGNIFICANCE: It is well known that hormonal stimulation of male fish induces the final maturation of spermatozoa. However, molecular and biochemical basis underlying hormone-induced changes in semen is unknown at present. This study for the first time reveals, using proteomic approach, that hormonal stimulation in addition to hydration of testis is accompanied by significant changes in seminal plasma proteins related mainly to immune and stress response, lipid metabolism, signal transduction and tissue remodeling. These changes are associated with gene expression and synthesis and secretion of seminal plasma proteins by reproductive tissues. Overall, our results provide a framework for understanding the molecular mechanism responsible for hormonal stimulation in the reproductive tract of fish males.
鲤鱼的激素刺激是一种常规做法，旨在增强精子的产生并控制配子的成熟。本研究旨在使用二维差异凝胶电泳比较对照和 Ovopel 诱导的雄性鲤鱼精液中的蛋白质组。Ovopel 诱导增加了精子体积、总精子数、精液渗透压和 pH 值，降低了精液蛋白浓度。总共鉴定出 36 个斑点（23 个上调和 13 个下调），这对应于 Ovopel 诱导后精液中差异丰富的 23 种蛋白质（p<0.05；倍数变化 1.2）。大多数蛋白质与免疫和应激反应有关，包括转运蛋白（hephaestin）、抗蛋白酶（胎球蛋白、α2-巨球蛋白、TIMP2）、补体成分（C3、补体因子 B/C2A）、凝血级联调节剂（纤溶酶原）、先天免疫反应调节剂，如 intelectin、ApoA 和 ApoE，以及组织蛋白酶/胱抑素系统，以及应激反应（烯醇酶 1）。此外，激素刺激似乎与参与脂质代谢、信号转导和组织重塑的蛋白质有关。我们的研究结果表明，激素刺激不仅伴随着睾丸的水合作用，还诱导与精子成熟和保护精子免受外源性激素处理引起的应激相关的精液蛋白的合成和分泌。意义：众所周知，鱼类雄性激素刺激可诱导精子的最终成熟。然而，目前尚不清楚激素诱导的精液变化的分子和生化基础。本研究首次使用蛋白质组学方法揭示，除了睾丸的水合作用外，激素刺激还伴随着与免疫和应激反应、脂质代谢、信号转导和组织重塑有关的精液蛋白的显著变化。这些变化与生殖组织的基因表达以及精液蛋白的合成和分泌有关。总的来说，我们的研究结果为理解鱼类雄性生殖道激素刺激的分子机制提供了一个框架。
