BREAD EAGER: Analysis of Bovine Epididymal and Chicken SST Sperm to Determine Proteins Involved in Sperm Preservation

超过92种爬行动物、两栖动物、鸟类和哺乳动物已经发展出将射精的精子储存在雌性生殖道中较长时间（数天至数年）而不降低其与卵子受精的能力的能力。然而，在大多数哺乳动物中，精子可以在附睾尾部保存长达15天，但在射精后只能存活大约24小时。这项研究的长期目标是确定精子可以长期储存在鸡的雌性生殖道和公牛的附睾中的机制，并利用这些知识开发一种无冷链系统，以允许家养农场动物人工授精，特别是在没有冷冻保存基础设施的发展中国家。深入了解创造附睾和鸡精液储存小管微环境的蛋白质将促进我们对这些蛋白质如何提高膜完整性以改善长期储存的理解。利用对附睾环境的新认识，可以延长无冷精子的储存时间，并可能在体细胞保存方面有其他应用。该奖项支持的研究将利用二维LC-MS/MS将牛附睾精子和射精精子的蛋白质谱与储存在雌性生殖道的鸡精子的蛋白质谱进行比较，并确定每组精子的线粒体活性。确定是否通过减少活动来延长寿命。然后将射精的公牛精子清洗并在含有附睾或鸡蛋白的培养基中培养，培养基的pH值为附睾精子的pH值，以确定精子动力学的变化。PI将招收一名硕士生作为该提案的组成部分，并将继续招收本科生和国际实习生进行研究培训并直接参与研究工作。这项研究的结果将发表在同行评议的期刊上，并通过科学会议传播。

Over 92 species of reptiles, amphibians, birds, and mammals have developed the ability to store ejaculated sperm in the female reproductive tract for extended periods of time (days to years) with no decrease in its ability to fertilize the ovum. However, in most mammals sperm can be maintained for up to 15 days in the cauda epididymis, but only live for approximately 24 hours after ejaculation. The long term goal of this research is to determine the mechanisms by which sperm can be stored long term in the female reproductive tract of chickens and in the epididymis of the bull, and to utilize the knowledge to develop a cold-chain-free system to allow artificial insemination of domestic farms animals, especially in developing countries that do not have the infrastructure for cryopreservation. A thorough understanding of the proteins that create the microenvironment of the epididymis and chicken semen storage tubules will advance our understanding of how these proteins may enhance membrane integrity to improve long term storage. Leveraging the new knowledge of the epididymal environment should enable cold-free sperm storage for extended periods, and may have other applications for somatic cell preservation. Studies supported by this award will utilize 2D LC-MS/MS to compare protein profiles of bovine epididymal sperm and ejaculated sperm to the protein profiles of chicken sperm stored in the female reproductive tract, and determine the mitochondrial activity in each group of sperm. To determine if increased longevity is accomplished through decreased activity. Ejaculated bull sperm will then be washed and cultured in media containing epididymal or chicken proteins at the pH of epididymal sperm to determine changes in sperm kinetics. The PI will recruit one Master's student to be an integral part of this proposal, and will continue to recruit undergraduate students and international interns for research training and direct involvement in the research efforts. Results from the study will be published in peer-reviewed journals and disseminated through scientific conferences.