Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Department of Gamete and Embryo Biology, Olsztyn, Poland.
Laboratory for Functional Genome Analysis (LAFUGA), Gene Center, Ludwig-Maximilians-Universität, Munich, Germany.
J Proteomics. 2014 Feb 26;98:218-32. doi: 10.1016/j.jprot.2014.01.005. Epub 2014 Jan 13.
The present study for the first time characterizes a diverse cohort of carp seminal and blood plasma proteins using the combination of protein fractionation by one-dimensional gel electrophoresis and high performance liquid chromatography electrospray ionization tandem mass spectrometry. Using this approach, we identified 137 proteins in carp seminal plasma and 88 proteins in carp blood plasma, most of which were newly identified in fish. Transferrin, serine proteinase inhibitors, apolipoproteins, complement C3 and Wap65 were present in high abundance in carp seminal plasma. In carp blood plasma, besides these proteins, immunoglobulins and macroglobulins were identified as major proteins. Comparative analysis of carp seminal and blood plasma proteome performed using 2D-DIGE revealed that in contrast to mammals the majority (1014 from 1240 spots) of carp seminal plasma proteins are blood proteins. Moreover, proteins more abundant in seminal plasma (99 from 1240 spots) were identified, including parvalbumin, isoforms of apolipoproteins, heat shock proteins, components of antioxidative system, matrix metalloproteinases, cathepsin D, enzymes of glycolysis and sperm structural proteins. These proteins are involved in the regulation of sperm motility, spermatogenesis, maintenance of sperm membrane lipid stability and antioxidant protection. This study enhances the basic knowledge concerning fish seminal plasma protein composition and their potential role in fish reproduction.
Proteins similar or identical to blood plasma components are important for male reproductive physiology. Comparative study of blood and seminal plasma is especially justified in fish. Using 2D-DIGE we indicated that, in contrast to mammals, in carp seminal plasma most proteins are common for blood and seminal plasma, which possibly is related to a lack of accessory glands in reproductive tract of most fish. The proteins present in higher abundance in seminal plasma can be related to physiology of fish male reproduction including regulation of sperm motility, spermatogenesis, maintenance of sperm surface composition and antioxidant protection. Application of proteomics analysis to identify carp seminal and blood plasma proteins significantly extends current knowledge regarding the composition of fish seminal and blood plasma proteins and their relationship to higher vertebrates. Moreover, proteomic profiling of carp seminal plasma appears to be helpful for further understanding of the role of fish seminal plasma proteins in male reproductive tract as well as for identification of novel biomarkers for sperm quality.
本研究首次使用一维凝胶电泳和高效液相色谱电喷雾串联质谱相结合的方法对鲤鱼精液和血浆中的蛋白质进行了多样化分析。采用这种方法，我们在鲤鱼精液中鉴定出 137 种蛋白质，在鲤鱼血浆中鉴定出 88 种蛋白质，其中大多数蛋白质是在鱼类中首次发现的。转铁蛋白、丝氨酸蛋白酶抑制剂、载脂蛋白、补体 C3 和 Wap65 在鲤鱼精液中含量丰富。在鲤鱼血浆中，除了这些蛋白质外，还鉴定出免疫球蛋白和巨球蛋白为主要蛋白质。使用 2D-DIGE 对鲤鱼精液和血浆蛋白质组进行比较分析表明，与哺乳动物不同，大多数（1240 个斑点中的 1014 个）鲤鱼精液蛋白质是血液蛋白质。此外，还鉴定出了在精液中含量更高的蛋白质（1240 个斑点中的 99 个），包括副肌球蛋白、载脂蛋白同工型、热休克蛋白、抗氧化系统成分、基质金属蛋白酶、组织蛋白酶 D、糖酵解酶和精子结构蛋白。这些蛋白质参与调节精子运动、精子发生、维持精子膜脂稳定性和抗氧化保护。本研究增强了我们对鱼类精液蛋白质组成及其在鱼类生殖中的潜在作用的基本知识。
与血液血浆成分相似或相同的蛋白质对雄性生殖生理很重要。在鱼类中，对血液和精液进行比较研究尤其合理。使用 2D-DIGE，我们指出，与哺乳动物不同，在鲤鱼精液中，大多数蛋白质是血液和精液共有的，这可能与大多数鱼类生殖道中没有附属腺有关。在精液中含量较高的蛋白质可能与鱼类雄性生殖生理有关，包括调节精子运动、精子发生、维持精子表面组成和抗氧化保护。应用蛋白质组学分析来鉴定鲤鱼精液和血浆蛋白质，显著扩展了我们对鱼类精液和血浆蛋白质组成及其与高等脊椎动物关系的现有知识。此外，鲤鱼精液蛋白质的蛋白质组学分析有助于进一步了解鱼类精液蛋白质在雄性生殖道中的作用，并有助于鉴定精子质量的新生物标志物。
