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Establishment of a developmental atlas and transgenetic tools in the ascidian Styela clava

Marine Life Science & Technology 2023年第4期5卷 435-454页

作者： Boyan Lin Wenjie Shi Qiongxuan Lu Takumi TShito Haiyan Yu Bo Dong Fang Zongxi Center College of Marine Life SciencesOcean University of ChinaQingdao 266003China Laoshan Laboratory Qingdao 266237China Department of Bioscience and Informatics Faculty of Science and TechnologyKeio UniversityYokohama 223-8522Japan MoE Key Laboratory of Evolution and Marine Biodiversity Ocean University of ChinaQingdao 266003China

The ascidian Styela clava is an ecologically important species that is distributed along coastal regions *** 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 ***,the standard developmental atlas and protocols and tools for molecular manipulation of this organism are *** this study,we established a standard developmental table and provided a web-based digital image resource for *** embryogenesis at each developmental stage from fertilized eggs to hatching larvae by utilizing confocal laser microscopy and 3D reconstruction *** 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 *** observed that the notochord cells in *** embryos did not produce an extracellular lumen like Ciona robusta,but showed polarized elongation behaviors,providing us an ideal comparative model to study tissue *** addition,we established a chemical-washing procedure to remove the chorion easily from the fertilized *** on the dechorionation technique,we further realized transgenic manipulation by electroporation and successfully applied tissue-specific fluorescent labeling in *** *** work provides a standard imaging atlas and powerful genetic tools for investigating embryogenesis and evolution using *** as a model organism.

关键词： Ascidian dechorionation Developmental atlas Styela clava Transgenic manipulation

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Injury-induced rapid activation of MAPK signaling in dechorionated eggs and larvae of the silkworm Bombyx mori

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Insect Science 2017年第2期24卷 248-258页

作者： Shi-Hong Gu Chien-Hung Chen Department of Biology National Museum of Natural Science Taichung Taiwan China Chung Hwa University of Medical Technol-ogy Tainan County Taiwan China

Previous study showed that diapause in Bombyx mori eggs can be ter- minated by dechorionation and that activation in the mitogen-activated protein kinase （MAPK）/extracellular signal-regulated kinase （ERK） in dechorionated cultured eggs is involved in diapause termination. In the present study, the possible mechanism underlying activation of ERK upon dechorionation was further investigated. Results showed that me- chanical injury of diapause eggs without medium incubation also resulted in rapid increase in the phospho-ERK levels and that injury increased the phospho-ERK levels at different stages of both diapause eggs and eggs in which diapause initiation was prevented by HC1. Effects of anaerobiosis on dechorionation-stimulated phospho-ERK levels showed that the mechanical injury itself but not the dramatic increase in oxygen uptake upon injury is in- volved in a rapid activation of ERK. Chemical anaerobiosis on dechorionation-stimulated phospho-ERK levels and the in vivo effect of anaerobiosis showed that the supply of oxygen also plays a role in ERK signaling. In addition, injury induced the phosphory- lation of c-jun N-terminal kinases （JNKs） and p38 kinase, components of two parallel MAPK pathways. A kinase assay showed a dramatic increase in .INK kinase activity in egg lysates upon injury. When newly hatched first instar larvae were injured, an increase in the phospho-ERK levels similar to that in dechorionated eggs was observed. From the results, we hypothesize that the injury-induced rapid activation of MAPK signaling, which serves as a natural signal for embryonic development, is related to diapause termination in dechorionated eggs.

关键词： Bombyx mori dechorionation embryonic diapause ERK phosphorylation injury MAPK

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