Microinjection of nucleic acids into zebrafish embroys

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The zebrafish, Danio rerio, has become one of the most widely used species for the study of vertebrate development and physiology due to its characteristics that facilitate embryological analyses and the fact that genetic approaches are possible. A number of methods rely on the microinjection of nucleic acids and gene constructs into fertilized zebrafish eggs to better understand biological processes in this species. For example, a simple, efficient, and specific method to investigate the function of genes in zebrafish is to "knock down" their function through injection of "morpholino oligonucleotides". Transgenic zebrafish that express the green fluorescent protein (GFP) gene in specific cell types under the control of tissue-specific promoters are useful to follow, in real time, the development, life, and death of specific cells and their response to environmental perturbations.    The Department of Biology of the University of Ottawa and the Center for Advanced Research in Environmental Genomics (CAREG) have recently developed what probably constitutes the largest concentration of Canadian researchers using zebrafish under the same roof.  This is the result of recent recruitments and the establishment of collaborations with existing researchers who have now adopted the zebrafish for part of their work. Thus, members of the Biology Department and CAREG are increasingly relying on the microinjection of nucleic acids into fertilized eggs for their research. The microinjectors currently available in the Department of Biology and which were purchased by the fish geneticists are fully used, creating a major bottleneck for all investigators. Therefore, we are requesting the purchase of one additional microinjector.    The understanding of biological processes in the zebrafish through manipulation of gene function will provide key findings that can be applied to other fish species as well as other classes of vertebrates.