Gouletsou Pagona G, Tsangaris George Th, Katsarou Eleni I, Bourganou Maria V, Barbagianni Mariana S, Venianaki Athina P, Bouroutzika Efterpi, Anagnostopoulos Athanasios K, Fthenakis George C, Katsafadou Angeliki I
Veterinary Faculty, University of Thessaly, 43100 Karditsa, Greece.
Proteomics Research Unit, Biomedical Research Foundation of the Academy of Athens, 11527 Athens, Greece.
Vet Sci. 2022 Dec 15;9(12):697. doi: 10.3390/vetsci9120697.
The objectives of the present work were to evaluate the semen of dogs by means of proteomics methods and to compare with proteomics results of the blood of the animals, in order to increase available knowledge on the topic and present relevant reference values for semen samples. Semen samples were collected from five Beagle-breed dogs. Reproductive assessment of the animals by means of clinical, ultrasonographic and seminological examinations confirmed their reproductive health. The sperm-rich fraction and the prostatic fraction of semen were processed for proteomics evaluation. LC-MS/MS analysis was performed by means of a LTQ Orbitrap Elite system. The technology combines high separation capacity and strong qualitative ability of proteins in biological samples that require deep proteome coverage. Protein classification was performed based on their functional annotations using Gene Ontology (GO). In blood plasma, semen sperm-rich fraction, and semen prostatic fraction, 59, 42 and 43 proteins, respectively, were detected. Two proteins were identified simultaneously in plasma and the semen sperm-rich fraction, 11 proteins in plasma and the semen prostatic fraction, and three proteins in the semen sperm-rich and prostatic fractions. In semen samples, most proteins were related to cell organization and biogenesis, metabolic processes or transport of ions and molecules. Most proteins were located in the cell membrane, the cytosol or the nucleus. Finally, most proteins performed functions related to binding or enzyme regulation. There were no differences between the semen sperm-rich fraction and prostatic fractions in terms of the clustering of proteins. In conclusion, a baseline reference for proteins in the semen of Beagle-breed dogs is provided. These proteins are involved mostly in supporting spermatozoan maturation, survival and motility, enhancing the reproductive performance of male animals. There appears potential for the proteomics examination of semen to become a tool in semen evaluation. This analysis may potentially identify biomarkers for reproductive disorders. This can be particularly useful in stud animals, also given its advantage as a non-invasive method.
本研究的目的是通过蛋白质组学方法评估犬精液，并与动物血液的蛋白质组学结果进行比较，以增加该主题的现有知识，并为精液样本提供相关参考值。从五只比格犬收集精液样本。通过临床、超声和精液学检查对动物进行生殖评估，确认它们的生殖健康。对精液的富含精子部分和前列腺部分进行蛋白质组学评估。使用LTQ Orbitrap Elite系统进行LC-MS/MS分析。该技术结合了高分离能力和对需要深度蛋白质组覆盖的生物样品中蛋白质的强大定性能力。基于使用基因本体论（GO）的功能注释进行蛋白质分类。在血浆、精液富含精子部分和精液前列腺部分中，分别检测到59种、42种和43种蛋白质。在血浆和精液富含精子部分中同时鉴定出两种蛋白质，在血浆和精液前列腺部分中鉴定出11种蛋白质，在精液富含精子和前列腺部分中鉴定出三种蛋白质。在精液样本中，大多数蛋白质与细胞组织和生物发生、代谢过程或离子和分子的运输有关。大多数蛋白质位于细胞膜、细胞质或细胞核中。最后，大多数蛋白质执行与结合或酶调节相关的功能。精液富含精子部分和前列腺部分在蛋白质聚类方面没有差异。总之，提供了比格犬精液中蛋白质的基线参考。这些蛋白质主要参与支持精子成熟、存活和运动，提高雄性动物的生殖性能。精液的蛋白质组学检查有可能成为精液评估中的一种工具。这种分析可能潜在地识别出生殖障碍的生物标志物。这在种畜中可能特别有用，也鉴于其作为一种非侵入性方法的优势。
