Zebrafish Facility

Managing PI: Holger Gerhardt

zebrafishZebrafish (Danio rerio) is a small aquatic vertebrate model that has already proven it invaluable power as a tool for the molecular deciphering of various biological processes. Hallmark features are a high fecundity (thousands of eggs on a weekly basis), rapid and extracorporeal development offering direct access to all embryonic stages, transparency during the first phases of embryonic development, and the availibility of numerous fluorescent reporter lines reporting specific internal organs or tissues and enabling easy visualization of these structures in a dynamic manner.

ZF CrisprIn addition, zebrafish are amenable to transient (morpholino oligonucleotide-mediated knockdown) or permanent (CRISPR/Cas9 mediated gene knockout or mutation, Tol2 transposon mediated transgenesis) genetic modulation of gene expression, as well as to pharmacological interference with protein function. Zebrafish embryos are thus particularly attractive models to identify or characterize regulatory molecules of diverse biological processes at a fast pace.

 

Fish Rooms

ZF roomThe CCB zebrafish facility comprises a 50ñsquare meter temperature- and light-controlled room equipped with a 5-rack and a 4-rack semi-closed recirculating housing system (ZebTec Multilinking Systems, Tecniplast), and a quarantine room equipped with a semi-closed recirculating Standalone table-top system (Tecniplast). The total housing capacity of the facility is up to 6000 zebrafish (ca 540 3.5L tanks). Adjacent laboratory rooms are equipped with injection set-ups for egg and embryo injection, incubators for embryo raising, fluorescence stereomicroscopy, and related small accessory equipment. For high-resolution live imaging a state-of-the-art confocal microscope (Leica SP8) is available in the Holger Gerhardt lab that is specially equipped with an incubation chamber to allow imaging of live embryos up to 24 hours and longer.

 
Chemicogenetics for gene and drug discovery

Genetic gain-of-function strategies (Tol2 transgenesis, mRNA overexpression), genetic loss-of-function approaches (morpholino antisense oligonucleotide based gene knockdown, CRISPR/Cas9 mediated gene knockout) and pharmacological inhibitor treatment approaches are applied for several projects at the CCB, along with high resolution fluorescent imaging and other phenotyping techniques. Combined with specific fluorescent labelling of tissues, structures or proteins in transgenic lines, these techniques allow specific screening of gene function in a certain tissue. Not only embryonic development is studied, but also disease models are available. At the CCB especially the hindbrain tumor model is of particular interest. In this model, tumor cells are injected in the hindbrain that develop into highly vascularized tumors within 3-5 days, and even develop metastases within this short time window.

 

 
References:

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  • Phng LK, Stanchi F, Gerhardt H. Filopodia are dispensable for endothelial tip cell guidance. Development. 140:4031-40, 2013.
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