Zebrafish Facility

The LCDS Zebrafish Facility is located in Building 560 on the Ft. Detrick campus and was established to provide the NCI-Frederick community with technical expertise, training, and support in the use of the zebrafish model system.  Advantages of using zebrafish as a model organism to study human disease states, such as cancer, include the amenability of the system to genetic approaches and the ability to directly visualize developmental processes.  Zebrafish have a short generation time (3-4 months) and can lay hundreds of eggs at weekly intervals.  Thousands of progeny can be collected from a single breeding pair, making biochemistry, genetic screening, and phenotypic assessments extremely feasible.  Unlike embryos of many other vertebrates, externally fertilized zebrafish can be observed and manipulated at all stages of development, and the embryos can be monitored at single-cell resolution for long periods of time.  This unique property of the zebrafish facilitates experimental techniques such as cell fate mapping, fluorescent tracer studies, time-lapse lineage analysis, and single-cell transplantation studies.  The optical clarity of zebrafish embryos also allows for morphogenetic events to be visualized during development, and a single animal can be analyzed for defects arising in multiple organs.  Morpholino (MO) antisense oligodeoxynucleotides can be used to block protein expression, and gene knockouts can be efficiently generated using CRISPR/Cas9 technologies.

The zebrafish are maintained in a Pentair Aquatic Habitats Z-Hab Duo system, and the facility contains three dissecting microscopes equipped with injectors and micro-manipulators.  A Nikon Eclipse Ni-E upright microscope is available for bright field as well as fixed and live cell fluorescent imaging of zebrafish embryos. For more information contact Christine Insinna-Kettenhofen (christine.kettenhofen@nih.gov).

Publications associated with Zebrafish Facility:

• Lu, Q.*, Insinna, C.*, Ott, C., Stauffer, J., Pintado, P.A., Rahajeng, J., Baxa, U., Walia, V., Cuenca, A., Hwang, Y.S., Daar, I., Lopes, S., Lippincott-Schwartz, J., Jackson, P.K., Caplan, S., and Westlake, C.J. Early steps in primary cilium assembly require EHD1/EHD3-dependent ciliary vesicle formation. Nat. Cell Biol. 17:228-240, 2015.
• Insinna, C.*, Lu, Q.*, Teixeira, I., Harned, A., Semler, E.M., Stauffer, J., Madgison, V., Tiwari, A., Kenworthy, A.E., Narayan, K., and Westlake, C.J., Investigation of the F-BAR domain PACSIN family uncovers membrane tubulation function in cilia assembly and transport. Nat Comm. doi: 10.1038/s41467-018-08192-9, 2019.
• Kota, P., Terrell, E. M., Ritt, D. A., Insinna, C., Westlake, C. J., and Morrison, D. K.  M-Ras/Shoc2 signaling modulates E-cadherin turnover and cell-cell adhesion during collective cell migration. Proc. Natl. Acad. Sci USA, 116:3536-3545, 2019.
• Walia, V.*, Cuenca, A.*, Vetter, M., Insinna, C., Perera, S., Lu, Q., Ritt, D.A., Semler, E., Specht, S., Stauffer, J., Morrison, D.K., Lorentzen, E., Westlake, C.J. Akt regulates a Rab11-effector switch required for ciliogenesis initiation. Dev. Cell. 50:229-246. 2019.
• Cuenca, A., Insinna, C., Zhao, H., John, P., Weiss, M.A., Lu, Q., Walia, V., Specht, S., Manivannan, S., Stauffer, J., Peden, A.A., Westlake, C.J. The C7orf43/TRAPPC14 component links the TRAPPII complex to Rabin8 for preciliary vesicle tethering at the mother centriole during ciliogenesis. J. Biol. Chem.  294:15418-15434, 2019.