Leys Sally P, Larroux Claire, Gauthier Marie, Adamska Maja, Fahey Bryony, Richards Gemma S, Degnan Sandie M, Degnan Bernard M
School of Integrative Biology, University of Queensland, Brisbane QLD 4072, Australia.
CSH Protoc. 2008 Dec 1;2008:pdb.prot5095. doi: 10.1101/pdb.prot5095.
INTRODUCTIONFertilization occurs internally in Amphimedon and embryos are brooded in multiple chambers throughout the adult. Each chamber contains a mixture of developmental stages, from egg to late ring stages (i.e., prehatch late embryos). At the end of embryogenesis, swimming parenchymella larvae emerge from the adult. After several hours in the water column, the larvae settle and metamorphose into juvenile sponges. This protocol details how to obtain Amphimedon larvae and post-larvae/juveniles as well as embryos. Once isolated, these biological stages can be used for a variety of molecular and cellular analyses.
引言
受精在仙女海绵体内进行,胚胎在成体的多个腔室中发育。每个腔室都包含从卵到晚期环胚阶段(即孵化前晚期胚胎)的不同发育阶段的混合体。在胚胎发育结束时,游动的实胚幼虫从成体中孵出。在水柱中漂浮数小时后,幼虫沉降并变态为幼年海绵。本方案详细介绍了如何获取仙女海绵的幼虫、幼虫后期/幼体以及胚胎。一旦分离出来,这些生物学阶段可用于各种分子和细胞分析。