EFFICACY OF MOUSE SPERM CRYOPRESERVATION

说明：（改编自申请人？s描述）革命性的进步 在基因组研究和创造基因特异性菌株的能力， 老鼠导致菌株数量呈指数级增长 可用于生物医学研究。冷冻和储存精子 有效地保存了大多数这些新创建的菌株，但现有的 这些方法并不能对所有品系的小鼠都有效。长期 本项目的目的是改进精子冷冻保存的方法， 可靠地恢复无病原体和生殖正常的后代。的 该项目的具体目标是： (1)建立有效的冷冻方案，以改善冷冻保存 不同近交系的精子各种添加和替换 现有冷冻保护剂制剂中的组分将在 窄范围的渗透压摩尔浓度，并测试其对 成功冷冻近交系和突变系精子。 (2)提高不同近交系配子体外受精的效果 菌株IVF的各种修改，如添加自由基 清除剂加入培养基中，并从培养基中除去透明质酸 将测试卵母细胞增强近交系受精的能力 配子 (3)确定病原体通过冻存精子传播的风险 在通过IVF或卵胞浆内精子注射恢复后代后 （ICSI）使用被感染男性的精子送到杰克逊实验室进行再衍生 以及进口一种将被诊断为存在 病原体 (4)提供动物生殖功能的定量评估 由冷冻的精子产生从冷冻保存的雌性和雄性动物中生产 精子将与ICR小鼠交配，以评估其生育能力，并将进行测试 对于生殖行为，测量血浆水平和体外释放 生殖相关激素，并评估精子发生。 神经内分泌机制可能是任何生殖缺陷的基础， 将被识别。

DESCRIPTION: (Adapted from Applicant?s Description) Revolutionary advancements in genome research and the ability to create genetically specific strains of mice are resulting in an exponential increase in the number of strains available for biomedical research. Freezing and storing sperm could most efficiently preserve the majority of these newly created strains, but existing methods do not work sufficiently well for all strains of mice. The long-term objective of this project is to improve methods of sperm cryopreservation for reliable recovery of pathogen free and reproductively normal offspring. The specific aims of this project are to: (1) Establish efficient freezing protocols for improved cryopreservation of sperm from different inbred strains. Various additions and substitutions of components in an existing cryoprotectant formulation will be made within a narrow range of osmolality and tested for their effect on the efficiency of successfully freezing inbred and mutant strain sperm. (2) Enhance in vitro fertilization (IVF) for gametes from different inbred strains. Various modifications of IVF such as addition of free radical scavengers to the culture medium and removal of the zona pellucida from oocytes will be tested for ability to enhance fertilization of inbred strain gametes. (3) Determine the risk of pathogen transmission through cryopreserved sperm upon recovery of offspring through IVF or intracytoplasmic sperm injection (ICSI) using sperm of infected males sent to The Jackson Laboratory for re-derivation and importation of a strain that will be diagnosed for presence of pathogens. (4) Provide quantitative assessment of reproductive function in the animals produced from frozen sperm. Females and males produced from cryopreserved sperm will be mated to ICR mice to evaluate their fertility and will be tested for reproductive behaviors, measured for plasma levels and in vitro release of reproduction-related hormones, and assessed for spermatogenesis. Neuroendocrine mechanisms that may underlie any reproductive deficits found will be identified.