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Zebrafish in genome research Can you spot the difference? What is a zebrafish? • Danio rerio • Small freshwater fish from South Asia. • 4 cm long when fully grown. • Common aquarium fish. • Very easy to look after. Image: Wikimedia commons/Marribio2 What is a model organism? • Non-human species widely studied to understand human disease. • Model organisms are used when experimentation using humans is unfeasible or unethical. • Can you think of a model organism? Types of model organism Genetic model organisms Experimental model organisms Genomic model organisms Good candidates for genetic analysis. Good candidates for research into developmental biology. Good candidates for genome research. Breed in large numbers. Produce robust embryos that can be easily manipulated and studied. Easy to manage genomes e.g. small genome size or limited number of repeats. Have short generation times so large scale crosses can be followed over several generations. Genome is similar to a human. Images: Wellcome Trust Sanger Institute Why use zebrafish? • Small size. • All major organs present within 5 days post fertilisation. • Short generation time (3-4 months). • Produces 300-400 eggs every 2 weeks. • Translucent embryos. • Lots of genome resources available. Image: TBC The zebrafish embryo brain ear eye heart swim bladder muscle block segments ~3.5 mm notochord Zebrafish and human disease • Zebrafish mutants have been produced to model human diseases such as: – Alzheimer's disease – congenital heart disease – polycystic kidney disease – Duchenne muscular dystrophy – malignant melanoma – leukaemia Forward screening for mutants ENU-treated male P +/+ female x +/M F1 +/+ x F2 +/+ (50%) +/M (50%) x F3 +/+ (25%) +/M (50%) M/M (25%) Reverse screening for mutants Potential human disease gene Exciting gene expression pattern Gene of interest Potential new player in developmental pathway Gene knockout Phenotype analysis The activity • Identify differences between the wildtype zebrafish and mutant zebrafish. • A glossary is provided to help you with scientific terms. Image: Rodrigo Young, University College London Flash cards & worksheets Answers Image 1 What’s the difference? Embryo B has no eye. Image: Rodrigo Young, University College London Image 2 What’s the difference? Fish B is a lighter, golden colour compared to fish A. Image: Keith C. Cheng, Penn State College of Medicine and Wellcome images Image 3 What’s the difference? The body of fry B is curved. If you look closely you’ll also see that its mouth is open. This is because it is unable to fully close its mouth as its muscles are too weak. Image: Elisabeth Busch, Wellcome Trust Sanger Institute and Lehtokari et al 2008, European Journal of Human Genetics Image 4 What’s the difference? The zebrafish embryos in picture B look paler and are not stained red. Image: Ana Cvejic, Wellcome Trust Sanger Institute Image 5 What’s the difference? There are bright green blobs in picture B. Image: Elisabeth Busch, Wellcome Trust Sanger Institute Image 6 What’s the difference? Embryo A has more blue dots than embryo B. The blue dots are stained neutrophils moving towards a wound on the zebrafish fin. Image: Ana Cvejic, Wellcome Trust Sanger Institute