The CBI Zebrafish platform
Zebrafish is a powerfull genetic and pharmacological model. It also bear amazing imaging properties as well as a diurnal life style. For all of these reasons it became a vertebrate model of choice in fields such as development, physiology and chronobiology. With the development of the CrispR techniques, it became possible to generate patient specific alleles, making it also a very promising model in the human genetics field. In addition, several systems are available in zebrafish for improving transgenesis (IsceI, Tol2), one of which is associated with a gateway cloning system (Tol2Kit (utah.edu)).
Equipment in the zebrafish platform
The facility contains two main rooms, one already containing approximately 1100 tanks and one that will be equipped in 2024 (CPER funding). The facility also contains a washing room equipped with a Calypso washing machine and three experimental rooms: -one that can be placed under a different light/cycle regime and can house adult zebrafish -one devoted to behavioural studies and one injection room. The behavior room is equipped with two ‘zebraboxes’ devoted to locomotor activity testing of zebrafish larvae and the injection room is equipped with three injection stations and two incubators. The facility also has a quarantine facility permitting importation of adult fish.
The present capacity of the facility is sufficient to accommodate the arrival of 1 new team using the zebrafish as their main animal model. In 2024, the platform will reach a capacity of 2200 tanks, which could accommodate another 2-3 CBI teams working heavily with the zebrafish model.
Situation in the local landscape, generation of tools within the TEFOR program
The zebrafish platform is unique in the local landscape being the only such facility located on a Toulouse research campus. It will begin to offer a variety of services, from simple housing of existing lines to the generation of new zebrafish (genome-editing and transgenesis). Establishing novel models for team outside the CBI will be organized through the national infrastructure Tefor, based in Saclay near Paris ( Tefor Paris-Saclay).
Teaching and training in developmental biology
The platform also serves as a teaching platform for Université Paul Sabatier providing embryos at levels from L1 to M1 and a research platform for M2R/M2Pro and ultimately PhD. In the past, when hosted in the CBD, before the CBI creation, the platform has served punctually as an ad hoc training centre for two Biopharm companies located in the Toulouse region (Département ADMET, Centre de Recherche Pierre Fabre, Castre; Département Recherche Exploratoire, Sanofi-Aventis, Toulouse).
The CBI Xenopus facility
Why Xenopus laevis?
Xenopus (Anurans) are most commonly used for anatomy, developmental biology, hormone studies, cellular and molecular biology and electrophysiology. The external fertilization and development of their eggs make them a prime candidate for studies involving biological and mechanical manipulations. Xenopus are very easy to maintain and can be induced up to two times a year to lay eggs with just a simple injection of hormone. They have very large eggs and the embryos develop quite quickly so microinjections, extracts and developmental analysis are all quite easy
It is fundamental to be sure about the quality of the production of animals to obtain pertinent scientific informations. It is why strict rules are observed in our Xenopus facilities in CBD. Xenopus housing is located at CBD in one room of about 35 meter square maintained at 18°C by air conditioning. 600 frogs are kept in an open circuit of carbon-filtered water. The advantage of this is that toxic ammonia is kept low and prevents diseases. Frogs are segregated by sex. A 12/12 light/dark cycle is maintained in the room.
Adult Xenopus laevis are carnivorous and eat submerged in water. Adults receive 2-3 times each week a commercial trout pelleted diet. After feeding, tanks are cleaned and the water totally changed (three times a week). X. laevis may be susceptible to disease in conditions of stress or when tank water are infected with Aeromonas and Pseudomonas bacteria (the cause of Red-Leg) or any gram negative or gram positive bacteria. Koizyme eliminates them by competitive exclusion and keep the Aeromonas, Pseudomonas and other pathogens at a very low level. To prevent bacteria over-proliferation Koizyme is added in each tank.
Our Xenopus facilities include a Xenopus husbandry. This is not frequent in Xenopus facilities. We have the complete control of our crossbreeding and of the quality of produced animal.
Use of Xenopus
At CBD two teams use Xenopus; the group of C. Leclerc and of E. Théveneau. Xenopus embryos and cells are used for developmental and cell biology experiments such as grafting, induction assays, cell signaling, in vivo and in vitro cell migration assays, traction forces measurements, activity measurements via biosensors (FRET), real-time analysis of cytoskeleton (actin, microtubules), calcium imaging as well as biochemistry studies. We also furnish and accommodate animals for other groups on the campus external to CBD:
Teaching in developmental biology
Xenopus is taught as a developmental model and used for practical work in 3 Licences modules
- Biologie & Environment: L2 UE biologie du développement animal and L2 UE for CAPES preparation
- Biologie cellulaire & physiologie with 2 L2 UE and 1 L3 UE
- Biologie prépa-concours (agronomy & veterinary) with 1 UE in L1 and 1 UE in L2
The CBI mouse zootechnics facility
The mouse zootechnics facility is located on the UPS campus. It belongs to the ANEXPLO Life Science Platform of Toulouse which includes 7 other sites from the Toulouse area with complementary technical skills on animal model creation and experimentation. It is open to academic and private laboratories. All projects are reviewed by the local ethics committee and the French Ministry of Research to ensure the respect of ethical principles.
The facility offers housing services for wild-type and genetically modified mice. It currently has a hosting capacity of 1500 cages. It is divided in several areas of differing health status:
- a SOPF area (2 rooms)
- a SPF area (5 rooms)
- an A2 area (1 room)
- a quarantine area (1 room).
- Bedding handling system, bottle processing system.
- Decontamination chamber, autoclave.
- 17 IVC racks, 7 air handling units, 7 changing stations.
- Experimentation/transgenesis room: PSM, micromanipulator, binocular magnifiers, anesthesia station.
- CO2 euthanasia chamber.
- All mice are housed in individually ventilated racks.
- The platform staff is in charge of bedding, food and water change, cleaning, well-fare monitoring, and breeding plan.
- Animals health and welfare status are monitored four times a year through veterinary inspections and analyses according to FELASA guidelines.
The Mouse behavioral core
The CBI Drosophila platform
Facility open to academic and non-academic groups.
- Michèle Crozatier, DR CNRS
- Luisa Di Stefano, CR CNRS
Permanent staff members:
- Julien Favier, T CNRS, Technical manager
- Amèlie Destenables, T CNRS
- Valérie Nicolas, T CNRS (30%)
Non-permanent : CDD (1day/week)
- CBI groups: 9
- Other academic: 2+University for TP
- Production of Drosophila food for all academic groups working with Drosophila in Toulouse (CBI, University TP, Restore, EDB,…)
- Genesis of Drosophila transgenic lines for all CBI groups (using AttP/AttB systems and CRISPR/Cas9 technology)
- Participation of facility members in setting up courses/ training programs
- Participation of facility members in setting up Master and PhD courses
- Student supervision: Training of Master’s students in Drosophila transgenesis.
- Apprenticeship training: Training of technicians in: Fly food preparation and Drosophila transgenesis
- Training for high school students
The CRCA EXPLAIN Apiary
The unit has an experimental apiary which has been registered as Regional Platform by the CNRS Regional Delegation.
The apiary ensures the control of genetic strains, handling and survey of pathologies, rational managements of populations, hive-product extraction, and offers laboratory rooms for behavioral experiments open to researchers willing to develop projects on bee behavior. Given the large surface surrounding the facilities, experiments in semi-natural conditions are also possible. It is a unique experimental station which was granted the status of regional platform by the Region midi Pyrenees and by the future Center of Integrative Biology.
Besides scientific activities, the apiary serves as a socio-economical support platform.
The CRCA Insect Raising facility
The unit has established raising facilities for several temperate and tropical species of arthropods exhibiting considerable differences in their biology including ants, crickets or spiders that are mostly used by the IVEP and CAB teams.
The facilities are also used to promote pedagogic initiatives because of the recognized educational benefits of keeping ants and ant nests in classrooms.