LEARNING OUTCOMES

The aim of the course is to provide students with the essential informations necessaries to carry out experiments on laboratory animals. Particularly, wide information on different animal models and their usefulness are given. Additionally, all necessary indications on law dispositions about animal research and on animal welfare are specified. Furthermore, the students will learn all methodologies currently in use to create mutant animals, including new scientific approaches. Specific examples are illustrated.

AIMS AND LEARNING OUTCOMES

The course is devoted to prepare students for experimental work on live animals. The students will be first taught of mouse gametogenesis, in order to understand later how the main biotechnologies, including transgenesis are carried out.

The course will also focus on the use of animals in biomedical research, to provide an introduction to the major areas of laboratory animal science, that contribute both to the humane health, to the animal care,  and to the quality of experimental results.

TEACHING METHODS

Frontal lessons and laboratory.

SYLLABUS/CONTENT

• Introduction, Historical notes and Legislation.
• Description of different type of animal models and their use in research, preclinical studies and for public health.
• Anatomy, physiology and pathology of laboratory animals.
• Animal welfare and 3R principle.
• Laboratory animals housing and breeding, and prevention of the risks.
• Animal models: outbred, inbred and mutant strains.
• Immunodeficiency, pathology, oncology and surgical models, and their experimental applications.
• In vivo analysis of the genic function: “Reverse” and “Forward” genetics
• Mouse molecular genetics: pronuclear and blastocyst microinjection
• Technology for generating transgenic animals, including mice and livestock.
• Mouse Embryonal Stem Cells and their use for mutant production, Knock-out and Knock-in: gene targeting strategies, homology recombination, positive/negative selection, BAC recombinering, ZFN, etc.
• Conditional expression: inducible promoters, transactivators, Tet-on/tet-off, cre/loxP, tamoxifen systems.
• Gene reporters and atlas of gene expression.
• Others technology: RNAi, etc.
• In vivo random mutagenesis: ENU and other chemical and physical mutagens.
• Insertional Mutagenesis: gene, enhancer and promoter trap, transposons.
• Examples of mutant models for the study of human diseases and implication of different genetic backgrounds on mutant phenotype.
• Cloning of endangered species
• Other vertebrate models: Zebrafish, Xenopus laevis, Gallus gallus
• Invertebrate models: Drosophila melanogaster as model organism for developmental biology studies and transgenic fly production; C. elegans and the RNA interference.

Laboratory training: mouse manipulation and necropsy; imaging in preclinical models and treatment evaluation; collection, microinjection and implant of mouse fertilized egg cells and blastocysts.

RECOMMENDED READING/BIBLIOGRAPHY

Handouts provided by the Professors