Maulana Tulus, Said Syahruddin, Arifiantini Raden Iis, Jakaria Jakaria, Gunawan Asep
Graduate School of Animal Production and Technology, Faculty of Animal Science, IPB University, Bogor, Indonesia.
Research Center for Applied Zoology, National Research and Innovation Agency, Bogor, Indonesia.
Front Vet Sci. 2025 Apr 10;12:1492135. doi: 10.3389/fvets.2025.1492135. eCollection 2025.
The characterization of sperm and seminal plasma proteins is essential for understanding bull fertility and optimizing reproductive success in buffalo bulls. Despite its importance, the reproductive proteomic of Toraya buffalo, an indigenous breed in Indonesia, remains largely unexplored. This study aimed to examine the seminal plasma and sperm proteins of Toraya buffalo to uncover those critical for reproductive functions. Semen samples were collected from eight Toraya buffalo bulls aged 4 to 10 years. Protein profiling was performed using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D-SDS-PAGE), followed by in-gel digestion and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Bioinformatics tools, including UniProt, PANTHER, DAVID, and STRING, were utilized to identify and annotate the detected proteins. This study successfully identified four key reproductive proteins: ADAM32 in seminal plasma and ZPBP, SPACA3, and CCDC136 in sperm. These proteins are essential for sperm motility, energy production, and acrosome formation, which are critical processes for fertilization. Additionally, many identified proteins were associated with metabolic pathways, particularly the tricarboxylic acid (TCA) cycle, which plays a fundamental role in energy supply for sperm function. In conclusion, this study offers the first comprehensive proteomic identification of seminal plasma and sperm proteins associated with reproductive functions in the Toraya buffalo. The findings highlight the presence of key proteins in sperm, including ZPBP, SPACA3, and CCDC136, as well as the identification of ADAM32 in seminal plasma, contributing to a deeper understanding of buffalo reproductive biology.
精子和精浆蛋白的特性对于理解水牛公牛的生育能力以及优化繁殖成功率至关重要。尽管其重要性，但印度尼西亚本土品种托拉亚水牛的生殖蛋白质组学在很大程度上仍未得到探索。本研究旨在检测托拉亚水牛的精浆和精子蛋白，以发现对生殖功能至关重要的蛋白。从8头年龄在4至10岁的托拉亚水牛公牛采集精液样本。使用一维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（1D-SDS-PAGE）进行蛋白质谱分析，随后进行胶内消化和液相色谱-串联质谱（LC-MS/MS）分析。利用包括UniProt、PANTHER、DAVID和STRING在内的生物信息学工具来鉴定和注释检测到的蛋白质。本研究成功鉴定出四种关键生殖蛋白：精浆中的ADAM32以及精子中的ZPBP、SPACA3和CCDC136。这些蛋白质对于精子活力、能量产生和顶体形成至关重要，而这些是受精的关键过程。此外，许多鉴定出的蛋白质与代谢途径相关联，特别是三羧酸（TCA）循环，其在为精子功能提供能量方面发挥着重要作用。总之，本研究首次全面鉴定了与托拉亚水牛生殖功能相关的精浆和精子蛋白。研究结果突出了精子中关键蛋白的存在，包括ZPBP、SPACA3和CCDC136，以及精浆中ADAM32的鉴定，有助于更深入地了解水牛生殖生物学。
