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.prot5184. doi: 10.1101/pdb.prot5184.
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, and they can produce high-quality embryos nearly year-round in the laboratory. After hatching from capsules, larval Ilyanassa can be maintained in culture, feeding on single-celled algae. The larvae will become competent to undergo metamorphosis after approximately 3 wk in culture. Metamorphosis can be induced artificially by treating with either the neurotransmitter serotonin or the nitric oxide synthase inhibitor 7-nitroindazole. Both of these reagents have been shown to induce metamorphosis in >75% of larvae within 48 h. This protocol describes the induction of metamorphosis in snail larvae.
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。它是螺旋动物发育以及不对称细胞分裂研究中尤为重要的模型。其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。伊利亚那萨成体很容易获得且便于在实验室饲养，并且它们几乎全年都能在实验室产出高质量的胚胎。从卵囊中孵化后，伊利亚那萨幼虫可以在培养环境中饲养，以单细胞藻类为食。幼虫在培养约3周后将具备变态能力。通过用神经递质血清素或一氧化氮合酶抑制剂7-硝基吲唑处理可人工诱导变态。这两种试剂都已被证明能在48小时内诱导超过75%的幼虫发生变态。本方案描述了蜗牛幼虫变态的诱导方法。
