Gupta Tripti, Mullins Mary C
Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine.
J Vis Exp. 2010 Mar 4(37):1717. doi: 10.3791/1717.
Over the last 20 years, the zebrafish has become a powerful model organism for understanding vertebrate development and disease. Although experimental analysis of the embryo and larva is extensive and the morphology has been well documented, descriptions of adult zebrafish anatomy and studies of development of the adult structures and organs, together with techniques for working with adults are lacking. The organs of the larva undergo significant changes in their overall structure, morphology, and anatomical location during the larval to adult transition. Externally, the transparent larva develops its characteristic adult striped pigment pattern and paired pelvic fins, while internally, the organs undergo massive growth and remodeling. In addition, the bipotential gonad primordium develops into either testis or ovary. This protocol identifies many of the organs of the adult and demonstrates methods for dissection of the brain, gonads, gastrointestinal system, heart, and kidney of the adult zebrafish. The dissected organs can be used for in situ hybridization, immunohistochemistry, histology, RNA extraction, protein analysis, and other molecular techniques. This protocol will assist in the broadening of studies in the zebrafish to include the remodeling of larval organs, the morphogenesis of organs specific to the adult and other investigations of the adult organ systems.
在过去20年里，斑马鱼已成为用于理解脊椎动物发育和疾病的强大模式生物。尽管对胚胎和幼体的实验分析广泛，且其形态已有详尽记录，但关于成年斑马鱼的解剖描述以及成年结构和器官的发育研究，连同针对成年个体的操作技术仍很欠缺。在从幼体到成体的转变过程中，幼体的器官在整体结构、形态和解剖位置上会发生显著变化。从外部看，透明的幼体发育出其特有的成年条纹色素模式和成对的腹鳍，而在内部，器官会经历大量生长和重塑。此外，具有双向分化能力的性腺原基会发育成睾丸或卵巢。本方案识别出了成年斑马鱼的许多器官，并展示了成年斑马鱼脑、性腺、胃肠系统、心脏和肾脏的解剖方法。解剖后的器官可用于原位杂交、免疫组织化学、组织学、RNA提取、蛋白质分析及其他分子技术。本方案将有助于拓宽斑马鱼研究范围，涵盖幼体器官的重塑、成年特有的器官形态发生以及成年器官系统的其他研究。
