Gharbiah Maey, Cooley James, Leise Esther M, Nakamoto Ayaki, Rabinowitz Jeremy S, Lambert J David, Nagy Lisa M
Department of Molecular and Cellular Biology, University of Arizona, Tucson, AZ 85721, USA.
Cold Spring Harb Protoc. 2009 Apr;2009(4):pdb.prot5186. doi: 10.1101/pdb.prot5186.
The marine gastropod Ilyanassa obsoleta is a long-standing and very useful model for studies of embryonic development. It is an especially important model for spiralian development, and for studies of asymmetric cell division. The embryos are amenable to classic embryological manipulation techniques, as well as a growing number of molecular approaches. Ilyanassa is also an important model for studies of metamorphosis, the ecology of parasitism, the effects of environmental contaminants on morphology and sexual function, and comparative neurobiology. Ilyanassa embryos are particularly well suited for RNA and protein localization studies because of the relatively large cells and favorable properties for imaging. This protocol describes how to fix and store Ilyanassa embryos and larvae for use in whole-mount in situ hybridization and immunohistochemical studies.
海洋腹足纲动物伊利亚那萨钝口螈是胚胎发育研究中一个长期且非常有用的模型。它是螺旋动物发育研究以及不对称细胞分裂研究中尤为重要的模型。这些胚胎适用于经典的胚胎学操作技术,以及越来越多的分子方法。伊利亚那萨钝口螈也是变态发育、寄生生态学、环境污染物对形态和性功能影响以及比较神经生物学研究的重要模型。由于细胞相对较大且具有利于成像的特性,伊利亚那萨钝口螈胚胎特别适合进行RNA和蛋白质定位研究。本方案描述了如何固定和保存伊利亚那萨钝口螈胚胎及幼虫,以用于全胚胎原位杂交和免疫组织化学研究。