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在雌性生殖道中储存射精精子的能力，并且不会降低其使卵子受精的能力。然而，在大多数哺乳动物中，精子可以在附睾尾中维持长达15天，但在射精后只能存活约24小时。这项研究的长期目标是确定精子可以在鸡的雌性生殖道和公牛的附睾中长期储存的机制，并利用这些知识开发一种无冷链系统，以允许对国内农场动物进行人工授精，特别是在没有冷冻保存基础设施的发展中国家。彻底了解的蛋白质，创造附睾和鸡精液储存小管的微环境，将推进我们了解这些蛋白质如何可以提高膜的完整性，以改善长期储存。 利用附睾环境的新知识，可以使精子在无冷条件下储存更长时间，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2D LC-MS/MS比较牛附睾精子和射精精子的蛋白质谱与储存在女性生殖道中的鸡精子的蛋白质谱，并确定每组精子中的线粒体活性。确定寿命的延长是否通过减少活动来实现。然后将射出的公牛精子洗涤并在含有附睾或鸡蛋白的培养基中在附睾精子的pH下培养，以确定精子动力学的变化。PI将招募一名硕士生作为本提案的组成部分，并将继续招募本科生和国际实习生进行研究培训并直接参与研究工作。 研究结果将发表在同行评审期刊上，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.