Response of Rhesus Sperm to Freezing: Recovery from Osmotic and Oxidative Stress

恒河猴精子对冷冻的反应：从渗透和氧化应激中恢复

• 批准号: 7449571
• 负责人: STUART A MEYERS
• 金额: $ 32.24万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7449571
• 关键词: Actins; Alcohol dependence; Artificial Insemination; Australia; Behavior; Biological Preservation; Biophysics; Cell Count; Cell membrane; Cell physiology; Cells; Cellular Stress; Cellular biology; Communicable Diseases; Condition; Contraceptive methods; Cryopreservation; Development; End Point; Fertilization in Vitro; Flow Cytometry; Fluorescence Microscopy; Freezing; Funding; Genotype; Goals; Grant; Human; Ice; Indiana; Individual; Infertility; Laboratories; Lead; Macaca; Macaca mulatta; Membrane; Methods; Mission; Modeling; National Center for Research Resources; Numbers; Oxidative Stress; Pharmaceutical Preparations; Physiological; Physiology; Population; Positioning Attribute; Process; Property; Proteomics; Reactive Oxygen Species; Recovery; Regenerative Medicine; Relative (related person); Research; Research Personnel; Semen Donor; Seminal fluid; Signal Transduction; Stress; Testing; Thinking; Time; Transgenic Organisms; Universities; Variant; Water; base; cell behavior; cell injury; cell motility; cold temperature; cryobiology; desire; embryonic stem cell; experience; genetic resource; human disease; loss of function; male; member; nervous system disorder; nonhuman primate; programs; response; somatic cell nuclear transfer; sperm cell; sperm function; success; vaccine development; Actins; Alcohol dependence; Artificial Insemination; Australia; Behavior; Biological Preservation; Biophysics; Cell Count; Cell membrane; Cell physiology; Cells; Cellular Stress; Cellular biology; Communicable Diseases; Condition; Contraceptive methods; Cryopreservation; Development; End Point; Fertilization in Vitro; Flow Cytometry; Fluorescence Microscopy; Freezing; Funding; Genotype; Goals; Grant; Human; Ice; Indiana; Individual; Infertility; Laboratories; Lead; Macaca; Macaca mulatta; Membrane; Methods; Mission; Modeling; National Center for Research Resources; Numbers; Oxidative Stress; Pharmaceutical Preparations; Physiological; Physiology; Population; Positioning Attribute; Process; Property; Proteomics; Reactive Oxygen Species; Recovery; Regenerative Medicine; Relative (related person); Research; Research Personnel; Semen Donor; Seminal fluid; Signal Transduction; Stress; Testing; Thinking; Time; Transgenic Organisms; Universities; Variant; Water; base; cell behavior; cell injury; cell motility; cold temperature; cryobiology; desire; embryonic stem cell; experience; genetic resource; human disease; loss of function; male; member; nervous system disorder; nonhuman primate; programs; response; somatic cell nuclear transfer; sperm cell; sperm function; success; vaccine development

DESCRIPTION (provided by applicant): Efficient preservation of genetic resources of the rhesus monkey (Macaca mulatta) is critical to the mission of the National Center for Research Resources. This application is aimed at the development of efficient macaque sperm cryopreservation and will aid in successful propagation of germplasm through artificial insemination and in vitro fertilization programs. Non-human primates (NHPs), especially genetically unique macaques, are important models of human disease with applications to infertility and contraception, infectious diseases and vaccine development, drug and alcohol addiction, neurological disorders, and regenerative medicine. Similar to human sperm, cryopreserved sperm from NHPs show variable cryoprotection success depending on the individual sperm donor. Semen donors are often selected based on successful cryopreservation results, however, many individual males, including rhesus males in research programs, have semen quality that fails to survive cryopreservation. This is likely to be particularly true concerning NHPs generated from embryonic stem cells, somatic cell nuclear transfer, and transgenics where propagation of a specific individual's genotype is desired. This grant analyzes some of the mechanisms that may cause poor cryoperformance and focuses on methods to uniformly cryopreserve these bimodal populations. Our central hypothesis is that an understanding of cryopreservation-associated physiological responses in sperm will lead to improvements in sperm cryosurvival. Our long-term goal is to develop an understanding of crucial mechanisms involved in cryopreservation of sperm in order to optimally preserve sperm from a variety of genotypically unique and valuable NHP males. Our approach will be to investigate the mechanisms of osmotic and oxidative cellular stress at the cell physiology level which includes membrane, cell signaling, and subcellular assessment of sperm function using fluorescence microscopy, flow cytometry, cryomicroscopy, and proteomics methods. As a team with members from UC Davis, Indiana University/Purdue University, and the University of Newcastle (Australia), we are well positioned to undertake the proposed research, because of our combined experience with fundamental cryobiology and sperm cell biology.

描述（由申请人提供）：有效保存恒河猴（Macaca Mulatta）的遗传资源对国家研究资源中心的任务至关重要。该应用旨在开发有效的猕猴精子冷冻保存，并有助于通过人工授精和体外受精计划成功地传播种质。非人类灵长类动物（NHP），尤其是遗传学独特的猕猴，是人类疾病的重要模型，在不育和避孕，传染病和疫苗发育，药物和酒精成瘾，神经系统疾病和再生医学中应用。与人类精子类似，NHP的冷冻保存精子取决于单个精子供体的可变性保护成功。经常根据成功的冷冻保存结果选择精液供体，但是，许多男性，包括研究计划中的恒河猴，具有无法在冷冻保存中幸存下来的精液质量。这对于从胚胎干细胞，体细胞核转移以及需要特定个体基因型传播的转基因产生的NHP尤其如此。该赠款分析了一些可能导致冷冻性能不良的机制，并专注于统一冷冻ression这些双峰种群的方法。我们的核心假设是，对精子中与冷冻保存相关的生理反应的理解将导致精子冷冻生存的改善。我们的长期目标是了解对精子冷冻保存涉及的关键机制的理解，以便从各种基因型独特且有价值的NHP雄性中最好地保留精子。我们的方法是研究细胞生理水平上渗透和氧化细胞应激的机制，包括膜，细胞信号传导以及使用荧光显微镜，流式细胞仪，冷冻显微镜检查和蛋白质组学方法对精子功能的亚细胞功能进行评估。作为与加州大学戴维斯分校，印第安纳大学/普渡大学和纽卡斯尔大学（澳大利亚）的成员的团队，我们在拟议的研究方面非常有能力进行拟议的研究，因为我们拥有基本的冷冻生物学和精子细胞生物学的综合经验。