Ambystoma Genetic Stock Center

钝口螈遗传资源中心

• 批准号: 10609292
• 负责人: Stephen Randal Voss
• 金额: $ 11.77万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10609292
• 关键词: Address; Administrative Supplement; Ambystoma; Ambystoma mexicanum; Behavioral; Biological; Biological Assay; Biology; Collection; Communities; Courtship; Cryopreservation; Culture Media; Data; Deposition; Derivation procedure; Disease; Embryonic Development; Employee; Female; Fertilization; Fertilization in Vitro; Freezing; Genetic; Harvest; Health; Human; Human Resources; Ice; International; Laboratories; Louisiana; Methods; Modeling; Monitor; Organ Model; Partner in relationship; Process; Protocols documentation; Reagent; Reproducibility; Research; Research Personnel; Resources; Sampling; Spermatids; Spermatogonia; Structure; Time; Training; Translating; Universities; Vas deferens structure; Work; agricultural center; experience; experimental study; genetic resource; male; organ regeneration; repository; sperm cell; sperm cryopreservation; sperm quality; sperm viability; success; tissue repair; undergraduate student

PROJECT SUMMARY The Ambystoma Genetic Stock Center (AGSC) is a P40 Research Resource that distributes laboratory axolotls (Ambystoma mexicanum) to researchers and educators nationally and internationally. The AGSC is currently working on a project to develop a reproducible protocol for cryopreserving axolotl sperm, which is needed to efficiently manage axolotl stocks within the AGSC and the broader axolotl community. Early results from this project found that axolotl spermatids are viable for only a short time after they are packaged into structures called spermatophores. This presents a challenge to develop a sperm cryopreservation protocol because there is a narrow window of time within which spermatophores can be harvested to obtain viable sperm. Additionally, spermatophores were found to be laid by males at various times of the day, more often at night and not during the workday. Temporal parameters for collecting spermatophores with viable sperm have not been defined because the mating process has never been rigorously monitored to know when spermatophores are deposited. Until these parameters are defined, it will not be possible to collect large numbers of viable spermatophores from which to develop and optimize methods for cryopreservation. There is critical need to develop a standard protocol for collecting spermatophores with viable sperm. There is also critical need to optimize and gain capacity for sperm freezing and thawing at the AGSC, and to integrate in vitro fertilization into the workflow. To address these needs, the AGSC will hire a full-time employee with previous experience in cryopreservation and axolotl biology. The new employee will work with undergraduate student workers to monitor axolotl courtship during the day and night for the purpose of recording behavioral data and collecting spermatophores. Spermatophores will be frozen and thawed using reagents and conditions recommended by the Aquatic Germplasm and Genetic Resources Center (AGGRC) at the Louisiana State University Agricultural Center and experiments will be performed to assay sperm viability and in vitro fertilization success. Protocols arising from the proposed work will be made publicly available. This administrative supplement will accelerate progress in developing an axolotl sperm cryopreservation protocol and building AGSC capacity to manage germplasm for the axolotl community. .

项目概要 Ambystoma 遗传库存中心 (AGSC) 是一个 P40 研究资源，负责分发 实验室蝾螈（Ambystoma mexicanum）向全国的研究人员和教育工作者提供 国际上。 AGSC 目前正在开展一个项目，开发可重复的协议 用于冷冻保存蝾螈精子，这是有效管理蝾螈种群所需的 AGSC 和更广泛的蝾螈社区。该项目的早期结果发现蝾螈 精子细胞在被包装成称为结构的结构后只能存活很短的时间 精细胞。这对制定精子冷冻保存方案提出了挑战 因为收集精子的时间窗口很窄 获得有活力的精子。此外，还发现雄性在不同的时间产下精囊。 一天中的某些时候，更常见的是在晚上而不是在工作日。时间参数 收集具有存活精子的精囊尚未定义，因为交配 从未对这个过程进行严格监控来了解精囊何时沉积。 在定义这些参数之前，不可能收集大量可行的 精细胞，从中开发和优化冷冻保存方法。有 迫切需要制定一个标准方案来收集具有存活精子的精细胞。 还迫切需要优化和获得精子冷冻和解冻的能力 AGSC，并将体外受精整合到工作流程中。为了满足这些需求， AGSC 将聘请一名具有低温保存和蝾螈经验的全职员工 生物学。新员工将与本科生一起监测蝾螈 白天和晚上的求偶行为，目的是记录和收集行为数据 精细胞。将使用试剂和条件冷冻和解冻精细胞 水产种质和遗传资源中心（AGGRC）推荐 路易斯安那州立大学农业中心和实验将进行分析 精子活力和体外受精成功率。拟议工作产生的协议将 公开发布。此次行政补充将加快发展进程 蝾螈精子冷冻保存方案和建设 AGSC 管理种质的能力 对于蝾螈社区来说。 。