CRYOPRESERVATION OF EJACULATED AND EPIDIDYMAL SPERM

目前在科学竞技场中发生的遗传学革命正在为理解基因的生物学功能和阐明它们在人类疾病中的作用创造新的范例。 这些类型的研究是有益的，因为它们可能产生新的诊断测试和治疗人类疾病。 突变小鼠的发生是建立基因功能及其在人类疾病中的作用的策略的重要组成部分。 因此，由国际科学界产生的突变小鼠的数量正以指数速度增加，并且目前将这些小鼠储存在储存库中的方法在成本和实用性方面并不理想。 一种可能的解决方案可能是小鼠精子的冷冻保存，但目前的方法是次优的和不一致的。 该提案旨在：（1）设计从这些研究者先前产生和研究的野生型和良好表征的转基因、基因敲除和ENU诱变小鼠品系射出小鼠精子的最佳方法;（2）优化/或开发从（1）中提到的小鼠品系冷冻保存小鼠精子的新方法;（3）将上述方法与子宫内或输卵管授精和/或体外受精方法和/或卵胞浆内精子注射结合使用，以获得尽可能高的活产幼崽率;和（4）使用免疫表型分析、组织病理学、行为基因表达（使用微阵列和/或SAGE分析）技术评价来自（3）的活产小鼠的正常性。

The genetics revolution currently occurring in the scientific arena is creating new paradigms in the understanding of the biological function of genes and elucidating their role in human disease. These types of studies are beneficial since they are likely to produce new diagnostic tests and treatments for human disease. The genesis of mutant mice is an important component of the strategy employed in establishing gene function and their role in human disease. Consequently, the number of mutant mice being generated by the international scientific community is increasing at an exponential rate and current methods of storing these mice in repositories are not ideal in terms of costs and practicality. One possible solution may be the cryopreservation of mouse sperm but current methodologies are sub-optimal and inconsistent. This proposal aims to: (1) devise optimal methods of ejaculating mouse sperm from wild-type and well-characterized strains of transgenic, gene knockout and ENU-mutagenized mice previously generated and studied by these investigators; (2) optimized/or develop new methods of cryopreservation of mouse sperm from strains of mice as mentioned in (1); (3) use the above methods in conjunction with intrauterine or oviductal insemination and/or in vitro fertilization methodologies and/or intracytoplasmic sperm injection so as to attain the highest possible rates of liveborn pups; and (4) evaluate the normality of liveborn mice from (3) using immunophenotyping; histopathology; behavioural gene expression (using microarray and/or SAGE analysis) techniques.