Ambystoma Genetic Stock Center

Specific Aim 1. Efficiently manage, produce, and distribute Mexican axolotls to enable biomedical research. The Ambystoma Genetic Stock Center (AGSC) at the University of Kentucky maintains a significant collection of Mexican axolotls in support of research and educational efforts nationally and internationally. The AGSC produces and distributes axolotl stocks using protocols that have been innovated and refined over four decades. The host institution is currently renovating new space and installing automated rearing equipment to double the size of the AGSC facility. Funding under this Aim will allow the AGSC to continue their central mission of providing axolotl stocks to users while implementing a new resource management plan. A new management plan is needed to meet increased demand for axolotl stocks and create new services, while ensuring animal health and long-term sustainability of the resource. One half of the newly renovated space will be used to increase the size of the adult population. This will increase the AGSC's capacity to fill large orders of juveniles and adults, which take considerable time and effort to rear. Also, by increasing production capacity, the AGSC will be able to innovate new products and services for the community. The other half of the newly renovated space will serve as a depository for transgenic lines that are deemed high priority by the AGSC advisory committee. Overall, accomplishment of this Aim will ensure distribution and long-term sustainability of axolotl resources to NIH-funded investigators. Specific Aim 2. Make axolotl information, protocols, and learning activities available to investigators and educators. The AGSC is more than a stock center; it also serves as an informatics hub for the community. Several times each week the PI, Associate Director, and Collection Curator respond to user questions, and every effort is made to provide answers to queries within 24 hours. The PI has considerable experience using the axolotl in a variety of research areas and maintains a computer server and website (Sal-Site) that distributes genome-based informatics to the research community. Under this Aim, the AGSC website will be administered and further developed to provide investigators with information about the collection, technical procedures, IACUC protocols, potential collaborators, and research findings. Also, AGSC staff will attend scientific meetings and make presentations to grow the user base. Finally, the AGSC will work with investigators to develop learning modules that focus on questions in regenerative biology and medicine. Accomplishment of this Aim will extend the axolotl knowledge base thus enhancing the animal resource. Specific Aim 3. Develop a custom microinjection service to assist investigators in the preparation of transgenic axolotl embryos. Several new transgenic approaches have been developed within the past few years that allow highly efficient and targeted modification of genomic sequence. While the axolotl will never be a genetic model organism, it is important to establish standardized methods for creating transgenic axolotls so that candidate gene functions can be manipulated and cellular fates and behaviors monitored. Preliminary data is presented to show that the AGSC is optimizing methods for: 1) Editing genes using TALENs, and 2) Knocking-in genes using the Tol2 system. Under this applied research Aim, protocols will be optimized to develop a custom service for injecting axolotl embryos. Investigators will have the option of visiting the AGSC and injecting embryos on-site, or they will submit prepared materials for injection by AGSC staff. The later option will bring transgenic axolotls to investigators that do not have sufficient infrastructure to house a breeding colony of axolotls for embryo production. Accomplishment of this Aim will speed development and increase access of transgenic axolotls for biomedical research.