Development of a Biology-Based Sperm Storage System

在大多数动物中，女性生殖道可以比精液传播者存储精子的时间更长。例如，与母鸡2-3周的储存相比，家禽精液在受精前通常只保存48小时。显然，女性生殖道具有人造延长器无法比拟的能力。基于支持性的初步结果，研究人员假设，在牛精子中添加特定的输卵管多糖可以延长精子的体外寿命。该奖项支持的研究旨在解决NSF面包挑战主题，即开发一种无冷链的人工授精过程。在发达国家，用液氮冷冻保存精子的方法彻底改变了牛精液的运输方式。但由于技术和基础设施的限制，冷冻保存技术对于发展中国家的小牧民来说是不切实际的。一种更实用的替代方案是允许有价值的雄性精液在环境温度下运输，最大限度地发挥有价值动物的遗传影响。哺乳动物输卵管能够延长精子寿命的特征尚不清楚，但有一种正在发展的范例，即输卵管多糖参与精子保留，并延长精子细胞的活力。这些假设将在3个具体目标中得到解决：1)鉴定与牛精子结合的糖链基序，并确定它们在输卵管中的位置；2)确定结合精子的输卵管多糖是否延长精子的寿命和在标准培养液(DmTALP)中的功能；以及3)确定输卵管多聚糖是否可以延长精子的寿命，并在优化的精液稀释液中在常温下发挥作用。除了研究结果，该项目还将培养一名博士后研究员和几名本科生，其中包括通过伊利诺伊大学的许多面向多元化的项目招收的未被充分代表的少数民族学生。一个交互式网站，讲授精液分析的重要性，每年将为175名学生开发并在实验室使用该网站。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。

In most animals, the female reproductive tract can store sperm for longer times than semen extenders. For example, poultry semen in extenders is usually only stored for up to 48 hours before insemination compared to the 2-3 week storage in the hen. Clearly, the female reproductive tract has capabilities unmatched by artificial extenders. Based on supportive preliminary results, the investigators hypothesize that adding specific oviduct glycans to bovine sperm will prolong sperm lifespan ex vivo. The studies supported by this award are designed to address the NSF BREAD Challenge topic of developing a cold-chain-free artificial insemination process. Sperm storage by cryopreservation in liquid nitrogen has revolutionized transportation of bovine semen in developed countries. But cryopreservation techniques are impractical for small herdowners in developing countries due to technical and infrastructure constraints. A more practical alternative would allow semen from valuable males to be transported at ambient temperature, maximizing the genetic impact of valuable animals. The characteristics of the mammalian oviduct that enable sperm to live an extended lifespan are unclear but there is a developing paradigm that oviductal glycans are involved in sperm retention and that the glycans extend sperm cell viability. These hypotheses will be addressed in 3 specific aims: 1) identification of glycan motifs that bind bovine sperm and determine their location in the oviduct; 2) determination of whether oviduct glycans that bind sperm prolong sperm lifespan and function in standard culture medium (dmTALP); and 3) determination of whether oviduct glycans can prolong sperm lifespan and function in an optimized semen diluent at normal room temperatures. In addition to the research findings, this project will train a postdoctoral fellow and several undergraduates, including underrepresented minority students recruited through the many diversity-oriented programs at the University of Illinois. An interactive web site that teaches the importance of semen analysis that will be developed and used in laboratories for a class of 175 students each year. Results from the study will be published in peer-reviewed journals and disseminated through scientific conferences.