The ascidian Styela clava is an ecologically important species that is distributed along coastal regions worldwide.It has a long history as a model animal for evolutionary and developmental biology research owing to i...The ascidian Styela clava is an ecologically important species that is distributed along coastal regions worldwide.It has a long history as a model animal for evolutionary and developmental biology research owing to its phylogenetic position between vertebrates and invertebrates,and its classical mosaic expression patterns.However,the standard developmental atlas and protocols and tools for molecular manipulation of this organism are inadequate.In this study,we established a standard developmental table and provided a web-based digital image resource for S.clava embryogenesis at each developmental stage from fertilized eggs to hatching larvae by utilizing confocal laser microscopy and 3D reconstruction images.It takes around 10 h for fertilized eggs to develop into swimming larvae and 20–30 min to complete the tail regression processes at the metamorphic stage.We observed that the notochord cells in S.clava embryos did not produce an extracellular lumen like Ciona robusta,but showed polarized elongation behaviors,providing us an ideal comparative model to study tissue morphogenesis.In addition,we established a chemical-washing procedure to remove the chorion easily from the fertilized eggs.Based on the dechorionation technique,we further realized transgenic manipulation by electroporation and successfully applied tissue-specific fluorescent labeling in S.clava embryos.Our work provides a standard imaging atlas and powerful genetic tools for investigating embryogenesis and evolution using S.clava as a model organism.展开更多
基金supported by the National Key Research and Development Program of China(2022YFC2601304,2022YFC2601302)the Science&Technology Innovation Project of Laoshan Laboratory(LSKJ202203002)+2 种基金the Taishan Scholar Program of Shandong Province,China(to BD)Database Construction was supported by the Research Institute of Marine Invertebrates(IKU2021-02)the Keio University Doctorate Student Grant-in-Aid Program from Ushioda Memorial Fund and JSPS KAKENHI Grant Number JP 22J22628,and Keio Gijuku Education with a Research-Adjusted Budget to TTS.
文摘The ascidian Styela clava is an ecologically important species that is distributed along coastal regions worldwide.It has a long history as a model animal for evolutionary and developmental biology research owing to its phylogenetic position between vertebrates and invertebrates,and its classical mosaic expression patterns.However,the standard developmental atlas and protocols and tools for molecular manipulation of this organism are inadequate.In this study,we established a standard developmental table and provided a web-based digital image resource for S.clava embryogenesis at each developmental stage from fertilized eggs to hatching larvae by utilizing confocal laser microscopy and 3D reconstruction images.It takes around 10 h for fertilized eggs to develop into swimming larvae and 20–30 min to complete the tail regression processes at the metamorphic stage.We observed that the notochord cells in S.clava embryos did not produce an extracellular lumen like Ciona robusta,but showed polarized elongation behaviors,providing us an ideal comparative model to study tissue morphogenesis.In addition,we established a chemical-washing procedure to remove the chorion easily from the fertilized eggs.Based on the dechorionation technique,we further realized transgenic manipulation by electroporation and successfully applied tissue-specific fluorescent labeling in S.clava embryos.Our work provides a standard imaging atlas and powerful genetic tools for investigating embryogenesis and evolution using S.clava as a model organism.
