Aix Marseille Univ, Avignon Université, CNRS, IRD, IMBE, Marseille, France.
Department of Genetics and Evolution, Faculty of Sciences, Institute of Genetics and Genomics in Geneva (IGe3), University of Geneva, Geneva, Switzerland.
Methods Mol Biol. 2021;2219:181-194. doi: 10.1007/978-1-0716-0974-3_11.
The Porifera are one of the best candidates as the sister group to all other metazoans. Studies on this phylum are therefore expected to shed light on the origin and early evolution of key animal features. Transcriptomic or genomic data acquired during the last 10 years have highlighted the conservation of most of the main genes and pathways involved in the development of the other metazoans. The next step is to determine how similar genetic tool boxes can result in widely dissimilar body plan organization, dynamics, and life histories. To answer these questions, three main axes of research are necessary: (1) conducting more gene expression studies; (2) developing knockdown protocols; and (3) reinterpreting sponge cell biology using modern tools. In this chapter we focus on the in situ hybridization (ISH) technique, needed to establish the spatiotemporal expression of genes, both on whole mount individuals and paraffin sections, and at different stages of development (adults, embryos, larvae, buds) of the homoscleromorph sponge Oscarella lobularis.
多孔动物门是最有可能成为所有后生动物姐妹群的生物之一。因此，对该门的研究有望揭示关键动物特征的起源和早期进化。在过去 10 年中获得的转录组或基因组数据强调了大多数主要基因和参与其他后生动物发育的途径的保守性。下一步是确定相似的遗传工具箱如何导致广泛不同的身体计划组织、动态和生活史。为了回答这些问题，需要进行三个主要的研究方向：（1）进行更多的基因表达研究；（2）开发基因敲低方案；（3）使用现代工具重新解释海绵细胞生物学。在本章中，我们重点介绍原位杂交（ISH）技术，该技术用于确定基因的时空表达，包括在整体个体和石蜡切片上，以及在同骨海绵 Oscarella lobularis 的不同发育阶段（成体、胚胎、幼虫、芽体）。
