Development of Germplasm Resources for Preservation of Aquatic Models

水生模型保存种质资源开发

基本信息

批准号：
8676954
负责人：
Terrence Robert Tiersch
金额：
$ 48.46万
依托单位：
LOUISIANA STATE UNIV AGRICULTURAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8676954
关键词：
Address Affect Animal Model Animals Back Biological Preservation Biomedical Research Biophysics Bladder Cell division Communities Computer Assisted Computers Confusion Coupled Coupling Cryopreservation Cryopreserved Cell Decision Making Deposition Development Disease Economic Inflation Educational workshop Embryo Engineering Ensure Equipment Equipment and Supplies Female Fertilization Fertilization in Vitro Fishes Flow Cytometry Freezing Future Genetic Research Genotype Germ Cells Goals Grant Health Human Laboratories Laboratory Research Learning Location Methods Modeling Modification National Center for Research Resources Nature Output Pathway interactions Performance Physiological Physiology Procedures Process Production Protocols documentation Reagent Recommendation Recovery Reporting Reproduction Research Resources Sampling Site Sperm Penetration Staging Standardization Swimming Terminology Testing Time Transgenic Organisms Zebrafish base conditioning design egg feeding genetic resource hatching improved mutant novel strategies offspring prevent repository sample collection sperm analysis sperm cell sperm quality success tool

项目摘要

DESCRIPTION (provided by applicant): In the past decade, laboratories around the world have produced tens of thousands of mutant, transgenic, and wild-type fish lines. Maintaining these valuable genotypes is expensive, risky, and beyond the capacity of even the largest stock centers. One of the major recommendations of the 2007 NCRR workshop 'Achieving High-Throughput Repositories for Biomedical Germplasm Preservation' was for performance of detailed studies to improve sperm cryopreservation for important aquarium fish models. Pursuant to this we propose to establish high-throughput processing of zebrafish sperm for stock centers and laboratories by extending a comprehensive cryopreservation pathway, and by intensifying efficiency and output to enable implementation of germplasm repositories. The Specific Aims are to: 1) broaden the pathway to improve quality and provide multiple access points for samples produced by current approaches, 2) intensify the pathway for automated processing and repository development including use of industrial modeling and decision-making tools, and 3) implement two scalable cryopreservation protocols; the large-scale high throughput method for Resource Center use and, reverse-engineered from it, a small to intermediate scale application for other research laboratories to back up their own lines or submit them to the Resource Center. This will improve quality, reduce variability, and increase efficiency. We propose to improve broodstock conditioning, gamete quality, fertilization and reproduction, and to accommodate the variety of approaches, levels of application, and current and future needs of the research community. This will yield two complementary approaches: one at an industrial scale using automated equipment and pooling of samples to yield the highest level of intensity and efficiency; the other, less intensive but highly reliable, designed or dissemination to other laboratories. The materials generated from this external pathway will feed into the central pathway and will provide an additional form of throughput based on the aggregate production from multiple locations. A high level of standardization will be necessary for both approaches to merge efficiently to deliver high-quality samples into large comprehensive germplasm repositories. Thus, we envision this renewal as a funnel to improve and adapt the wide breadth of technical approaches available for zebrafish cryopreservation, and to focus these activities for repository development that can be eventually coupled with commercially available high-throughput equipment, supplies, and reagents specifically optimized for aquarium fish. This approach is practical and extremely feasible. This renewal proposal builds directly on the success of our established pathway and looks to couple efficiently with future advances.

描述（由申请人提供）：在过去的十年中，世界各地的实验室已经生产了数以万计的突变型、转基因型和野生型鱼品系。维持这些有价值的基因型是昂贵的、有风险的，甚至超出了最大的库存中心的能力。 2007 年 NCRR 研讨会“实现生物医学种质保存的高通量存储库”的主要建议之一是进行详细研究，以改进重要观赏鱼模型的精子冷冻保存。据此，我们建议通过扩展综合冷冻保存途径，并通过提高效率和产量以实现种质资源库的实施，为储备中心和实验室建立斑马鱼精子的高通量处理。具体目标是：1) 拓宽提高质量的途径，并为当前方法生成的样本提供多个访问点，2) 强化自动化处理和存储库开发的途径，包括使用工业建模和决策工具，以及 3 ) 实施两个可扩展的冷冻保存协议；供资源中心使用的大规模高通量方法，并对其进行逆向工程，为其他研究实验室提供中小型应用程序，以备份自己的生产线或将其提交给资源中心。这将提高质量、减少变异性并提高效率。我们建议改善亲鱼调理、配子质量、受精和繁殖，并适应研究界的各种方法、应用水平以及当前和未来的需求。这将产生两种互补的方法：一种是在工业规模上使用自动化设备和样本池，以产生最高水平的强度和效率；另一种是在工业规模上使用自动化设备和样本池，以产生最高水平的强度和效率。另一种是强度较低但高度可靠的，是设计或传播给其他实验室的。从该外部路径产生的材料将进入中央路径，并将根据多个地点的总产量提供另一种形式的吞吐量。高水平的标准化对于这两种方法的有效融合是必要的，以便将高质量的样本传送到大型综合种质资源库中。因此，我们将这种更新设想为一个漏斗，以改进和适应可用于斑马鱼冷冻保存的广泛技术方法，并将这些活动集中于存储库开发，最终可以与商业上可用的高通量设备、用品和试剂结合起来针对观赏鱼进行了优化。这个办法是切实可行的，也是极其可行的。这一更新提案直接建立在我们既定途径的成功基础上，并希望与未来的进步有效结合。