EFFICACY OF MOUSE SPERM CRYOPRESERVATION

描述：（改编自申请人的描述）革命性的进步 基因组研究和创造遗传特异性菌株的能力 小鼠导致菌株数量呈指数增长 可用于生物医学研究。冷冻和储存精子最有可能 有效地保存了大多数这些新创建的菌株，但现有的菌株 这些方法并非对所有品系的小鼠都有效。长期来看 该项目的目标是改进精子冷冻保存方法 可靠地恢复无病原体且繁殖正常的后代。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 来自不同近交系的精子。各种添加和替换 现有冷冻保护剂配方中的成分将在 狭窄的渗透压范围并测试了它们对效率的影响 成功冷冻近交系和突变株精子。 (2) 增强不同近交系配子的体外受精（IVF） 菌株。 IVF 的各种修改，例如添加自由基 培养基中的清除剂并去除透明带 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 通过体外受精或胞浆内单精子注射恢复后代后 (ICSI) 使用被送往杰克逊实验室的受感染男性的精子进行重新衍生 以及输入将被诊断是否存在的菌株 病原体。 (4) 提供动物生殖功能的定量评估 由冷冻精子产生。冷冻保存的雌性和雄性 精子将与 ICR 小鼠交配以评估其生育能力并进行测试 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。 可能导致任何生殖缺陷的神经内分泌机制 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.