Animals

The zebrafish model arrived at the CBD in November of 2001 with the recruitment of a research team directed by Patrick Blader. The choice of installing this new model aimed at complementing the chicken, mouse and Xenopus models already used at the CBD with a “forward genetic” vertebrate model. While the initial installation of the zebrafish aquarium facility provided for 5 tank racks (maximum capacity 3500 adults animals), in the spring of 2007 the installation was expanded to its present capacity of 12 tank racks (approximate capacity 10000 adult zebrafish); at present, the number of fish housed in the CBD zebrafish facility varies between 6000-8000 adult animals, including approximately 35-40 transgenic and 25-30 mutant lines.

A transgenesis platform composed of three microinjection stations and a cell transplantation rig are associated with the animal facility. At present capacity, the zebrafish facility offers a platform for three research groups at the CBD (Blader, Vandel and Soula) and also to other groups on and around the UPS campus (Gleizes, Vallet). In keeping with the expansion of capacity of adults, a larval raising facility better adapted to the new capacity was constructed in 2012. Finally, despite the recent expansion of adult capacity the CBD zebrafish facility is already saturated, primarily due to the increase in the number of user in each of the three CBD teams using the facility. A strategy has been put in place to increase the floor space of the facility that would permit the installation of 5-8 new tank racks.

The zebrafish platform is unique in the local landscape being the only such facility located on a Toulouse research campus. 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. Finally, 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).

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

Xenopus housing

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.

Care

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.

Xenopus husbandry

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 ABC Zootechnics Facility is located on the UPS campus. Its management is shared by two laboratories (Centre de Biologie du Développement and Centre de Recherches sur la Cognition Animale). It is open to academic and private laboratories. All projects are reviewed by the FRBT ethical committee to ensure the respect of ethical principles.

The ABC Zootechnics Facility 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. The ANEXPLO platform received the IBiSA label and is certified ISO9001/2008. Services and equipment

The ABC facility offers housing services for mice, genetically modified or not. It has a hosting capacity of 850 cages. It is divided in two areas of differing health status:

• A conventional zone (600 cages).
• A quarantine (150 cages).

Equipments

• CO2 euthanasia apparatus meeting european guidelines.
• Sterilization equipment for chemical decontamination and autoclaving.
• The facility also includes an experimentation room containing:
• Anesthesia apparatus.
• Binoculars for dissections of tissue samples.
• CO2 incubator
• Fridge and freezer

Services

• Mice in quarantine are housed in individually ventilated racks.
• In conventional zones, mice are housed in cages with filtered lids.
• Animals health and welfare status are monitored twice a year through veterinary inspections and analyses according to FELASA guidelines

• https://anexplo.genotoul.fr/comportement/
• http://cbi-toulouse.fr/fr/mouse-behavioral-phenotyping-core

Four research groups (around 60 persons) in the CBD are working on Drosophila development. The CBD’s “fly platform” is an organized infrastructure that provides facilities for the daily work of  all these persons. In addition, the CBD “fly platform” provides facilities for the three new Drosophila’s groups that recently joined the LBMCP and one group from the CRCA. The “fly platform” has helped to develop a strong contribution of CBD’s researchers in the undergraduate teaching program.

To maintain and culture flies, the CBD has invested in common facilities which include a fly food preparation kitchen with a large production Finally, two microinjection stations are available for generating transgenic flies.

An essential resource is the mutant stock collection; which includes up to 2000 fly lines. This collection includes mutants stocks generated locally through our specific research programs (for example, a Toulouse-generated collection of about 250 P-lacZ or P-Gal4 enhancer trap lines on the X chromosome). In addition, many fly lines that are available from international stock centers are maintained also locally and are freely exchanged between the teams (it includes Gal4 and UAS transgenic lines).

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. Lucie Hotier is actively engaged in advising local beekeepers with respect to bee pathologies and rational handling of bee colonies.

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. Gérard Latil has developed over the years a specialized and unique knowledge on insect raising methods. As a result he is regularly consulted to provide advice on the subject to other biology laboratories and public organizations in France. The facilities are also used to promote pedagogic initiatives because of the recognized educational benefits of keeping ants and ant nests in classrooms.