Microinjection of nucleic acids into zebrafish embroys

斑马鱼Danio rerio已成为脊椎动物发育和生理研究中应用最广泛的物种之一，因为它的特性有利于胚胎学分析，而且遗传方法是可能的。许多方法依赖于将核酸和基因结构显微注射到受精的斑马鱼卵中，以更好地了解该物种的生物学过程。例如，研究斑马鱼基因功能的一种简单、有效和特异的方法是通过注射“吗啉寡核苷酸”来“敲除”它们的功能。在组织特异性启动子的控制下，转基因斑马鱼在特定的细胞类型中表达绿色荧光蛋白(GFP)基因，有助于实时跟踪特定细胞的发育、生活和死亡以及它们对环境扰动的反应。最近，渥太华大学生物系和环境基因组学高级研究中心(Careg)开发出了可能构成加拿大研究人员在同一屋檐下使用斑马鱼的最大集中度。*这是最近招聘的结果，并与现有研究人员建立了合作，这些研究人员现在已经将斑马鱼作为他们工作的一部分。因此，生物系的成员和卡雷格越来越依赖于将核酸显微注射到受精卵中进行研究。目前生物系现有的、由鱼类遗传学家购买的微型注射器已全部用完，给所有调查人员造成了重大瓶颈。因此，我们请求再购买一台微型注射器。因此，通过操纵基因功能来理解斑马鱼的生物过程将提供关键发现，这些发现可以应用于其他鱼类以及其他类别的脊椎动物。

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.