Zebrafish Core

Seeinstructions):

Seeinstructions): The Zebrafish Core (Core B) will provide direct support to Core A and all projects incorporating studies utilizing transgenic zebrafish. The Core is directed by Dr. Michael Parsons, an experienced investigator in zebrafish genetics, and will be supported by additional zebrafish technicians by the second year. The following Specific Aims will be pursued: First, to provide centralized embryo microinjection services for generating novel transgenic lines; second, to provide expert daily care for each of the transgenic fish to be generated in this proposal, including feeding and tank maintenance, breeding and genotyping services; third, to provide state-of-the-art imaging of adult and embryonic zebrafish, including confocal time-lapse microscopy. Core B will be responsible for the generation, breeding and maintenance of all wild-type and genetically engineered zebrafish stocks to be utilized by the four projects and Core A. This will include daily production of wild-type embryos, and embryo microinjection of Tol2 transposon-based elements for high- efficiency transgene integration. Assisted by the use of bicistronic transgenic vectors, Core B will also screen for germline transmission and select F1 founders for generating independent lines. Core B technicians will also be responsible for daily feeding, tank cleaning, water quality testing. Core B will also co-ordinate breeding of adult fish, both for maintenance of existing lines and for crossing of lines onto other transgenic backgrounds. As needed, fin clips from anesthetized fish will be provided to investigators for PCR genotyping. Finally, Core B will take advantage of Dr. Parsons' extensive experience with confocal imaging of living fish, allowing delineation of even subtle phenotypes associated with pancreatic expression of coding and non-coding candidate cancer genes. Core B is housed in a newly completed 1500 sq. ft. Shared Zebrafish Facility. In addition to space for embryo injection and embryo sorting, the facility is outfitted with a 34-rack Aquatic Habitats continuous flow system with continuous water quality monitoring and automated dosing for maintenance of constant pH and salinity. The system has a capacity for -1400 2.5 liter tanks, and can house over 30,000 adult fish. RELEVANCE (See instructions): Pancreatic cancer represents one of the most deadly human malignancies, with five year survival rates of less than 5% and no change in this figure over the past four decades. By determining the genetic basis for this disease, our program will generate clinically relevant information that is likely to directly impact on strategies for chemoprevention, early detection and treatment.