Prpic Nikola-Michael, Schoppmeier Michael, Damen Wim G M
Institute for Genetics, Evolutionary Genetics, University of Cologne, 50674 Köln, Germany.
CSH Protoc. 2008 Oct 1;2008:pdb.prot5067. doi: 10.1101/pdb.prot5067.
INTRODUCTIONThe spider Cupiennius salei, commonly known as the American wandering spider, is a particularly useful laboratory model for embryological studies because of the availability of tools to study and manipulate its embryonic development. Cupiennius is used to study axis formation, segmentation, appendage development, neurogenesis, and silk production. These studies contribute to our understanding of the evolution of these processes, but they also help us to understand the origin and diversification of evolutionary novelties. Comparisons between spiders and insects can show the degree of conservation and divergence of developmental mechanisms during arthropod evolution. Any embryological feature conserved between spiders and insects is likely to represent an ancestral feature for arthropods. Comparative molecular embryological work in insects and spiders should eventually allow us to define a molecular archetype for the phylum Arthropoda. This in itself will be a necessary cornerstone for comparing the different metazoan phyla, including chordates. This protocol describes the collection and fixation of embryos from C. salei. The fixed embryos can be stored at -20°C for prolonged periods and used for in situ hybridization, in studies of apoptosis using terminal deoxynucleotidyl-transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL), and for immunohistochemistry.
引言
蜘蛛Cupiennius salei，通常被称为美洲游走蛛，是胚胎学研究中一种特别有用的实验模型，因为有研究和操纵其胚胎发育的工具。Cupiennius被用于研究轴的形成、体节形成、附肢发育、神经发生和吐丝。这些研究有助于我们理解这些过程的进化，同时也有助于我们理解进化新特征的起源和多样化。蜘蛛和昆虫之间的比较可以显示节肢动物进化过程中发育机制的保守程度和差异程度。蜘蛛和昆虫之间任何保守的胚胎学特征都可能代表节肢动物的祖先特征。对昆虫和蜘蛛进行比较分子胚胎学研究最终应能让我们确定节肢动物门的分子原型。这本身将是比较包括脊索动物在内的不同后生动物门的必要基石。本方案描述了从C. salei采集和固定胚胎的方法。固定后的胚胎可在-20°C长期保存，并用于原位杂交、使用末端脱氧核苷酸转移酶介导的dUTP-地高辛标记（TUNEL）进行凋亡研究以及免疫组织化学。
