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.prot5183. doi: 10.1101/pdb.prot5183.
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 adults are readily obtainable and easy to keep in the laboratory. Although the normal spawning season for Ilyanassa is during early summer, they can produce high-quality embryos nearly year-round in the laboratory. Snails collected in the late fall, winter, or spring can be induced to deposit zygotes before the natural spawning season by warming them to room temperature, and snails collected before the natural spawning season can be made to postpone zygote deposition until needed (up to at least 6 mo) by maintaining them in tanks in a cold room at 4 degrees C-8 degrees C. This protocol describes how to induce embryo production in Ilyanassa snails, collect the embryos, and rear them to the stage required for study.
海洋腹足纲动物过时岩螺是研究胚胎发育的一个长期且非常有用的模型。它是螺旋动物发育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。过时岩螺也是变态发育、寄生生态学、环境污染物对形态和性功能的影响以及比较神经生物学研究的重要模型。过时岩螺成体很容易获得且便于在实验室饲养。虽然过时岩螺的正常产卵季节在初夏,但在实验室里它们几乎全年都能产出高质量的胚胎。在深秋、冬季或春季采集的蜗牛,通过将其升温至室温可诱导它们在自然产卵季节之前产下受精卵;而在自然产卵季节之前采集的蜗牛,通过将它们置于4摄氏度至8摄氏度冷藏室的水箱中饲养,可使其推迟受精卵的产出,直至需要时(至少长达6个月)。本方案描述了如何诱导过时岩螺产出胚胎、收集胚胎并将其饲养到研究所需的阶段。