- Zebrafish core facility
Zebrafish Core Facility
The Zebrafish Core Facility (ZCF) has existed since 2012. It is a licensed breeding and research facility (PL14656251 – registry of the District Veterinary Inspectorate in Warsaw; 064 and 051 – registry of the Ministry of Science and Higher Education). The establishment of the facility was to introduce a new vertebrate model in research conducted at IIMCB.
Zebrafish, a small (3 cm - 5 cm) freshwater tropical fish is an excellent model in biomedical research due to high genetic similarity to human, transparency of embryos, a very short reproduction cycle, an access to experimental manipulation, a large mutant/transgenic collection and low cost of maintenance. Furthermore, zebrafish as a lower vertebrate are an attractive alternative to mice and rats and can be used for implementation of the “3R” principles (reduction, replacement, and refinement). In 2013, approximately 6000 fish (30 lines) were kept in ZCF in 300 tanks (50 tanks in quarantine and 250 tanks in the main system). Presently, zebrafish stock collection contains more than 16 000 fish including wild type lines, almost 50 mutant lines and more than 30 transgenic lines (summary presented in the Figure 1). In addition to the lines listed in the Table 1, in ZCF are housed many of lines with affected genes involved in the mTOR signaling pathway, mitochondrial processes, the heart development or bearing mutations related to neurodegenerative disorders. Transgenic lines available in the ZCF are used for studying cellular processes and allow gain insights into the mechanisms of diseases in vertebrates. Many of the mutants were generated using methods based on engineered endonucleases such as transcription activator-like effector nucleases (TALENs) or the bacterial type II clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system. At present eight research groups from IIMCB use fish and equipment resources of the facility. In 2016, ZCF also served external users including research groups from the University of Warsaw, the Medical University of Warsaw, the Warsaw University of Life Sciences, the Nencki Institute of Experimental Biology, the University of Warmia and Mazury in Olsztyn.
Maintaining such a large number of fish would not be possible without a suitable infrastructure. The aquatic systems are manufactured by Tecniplast and now fish are housed in 970 tanks (6 independent, automated systems). Moreover, ZCF is equipped with incubators, microscopes, injectors, and a thermocycler. To enable the precise injection a needle puller, a beveler, and a microforge are used to prepare micro-needles. ZCF users have at their disposal a room dedicated for behavioral tests on zebrafish. The room is equipped with a small system for housing adult fish, ensuring an acclimatization period to allow for the stabilization of parameters before behavioral tests and two automated systems which allows observations and tracking of larval and adult zebrafish. In 2016 ZCF has started to freeze the sperm what ensures the survival of valuable lines and reduces the number of tanks used in the facility. Colony health is strictly monitored by veterinarian.
Scientists who use zebrafish for research purposes are obliged by law (ACT of 15 January 2015 on the protection of animals that are used for scientific or educational purposes) to possess appropriate qualifications to work with an animal model. All of the research and breeding activities at ZCF are carried out in compliance with fundamental ethical principles and in compliance with ACT of 15 January 2015 and European and international guidelines on animal welfare, including Directive 2010/63/EU on the protection of animals that are used for scientific purposes and the guidelines and recommendations of the Federation of European Laboratory Animal Science Associations (FELASA).