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Ciona intestinalis as an emerging model organism: its regeneration under controlled conditions and methodology for egg dechorionation

Ciona intestinalis as an emerging model organism: its regeneration under controlled conditions and methodology for egg dechorionation

作     者:LIU Li-ptng XIANG Jian-hai DONG Bo NATARAJAN Pavanasam YU Kui-jie CAI Nan-er 

作者机构:Key Laboratory of Experimental Marine Biology Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China College of Aqua-Life Science and Technology Shanghai Fisheries University Shanghai 200090 China Department of Aquatic Biology and Fisheries University of Kerala Trivandrum 695034 India 

出 版 物:《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 (浙江大学学报(英文版)B辑(生物医学与生物技术))

年 卷 期:2006年第7卷第6期

页      面:467-474页

核心收录:

学科分类:0710[理学-生物学] 07[理学] 09[农学] 

基  金:Project supported by the Hi-Tech Research and Development Pro-gram (863) of China (No. 2003AA603440) and the Knowledge In-novation Program from the Chinese Academy of Sciences (No.KZCX2-211) 

主  题:Cona intestinalls Model organism Laboratory culture Embryogenesis 

摘      要:The ascidian Ciona intestinalis is a model organism of developmental and evolutionary biology and may provide orucial clues concerning two fundamental matters, namely, how chordates originated from the putative deuterostome ancestor and how advanced chordates originated from the simplest chordates. In this paper, a whole-life-span culture of C. intestinalis was conducted. Fed with the diet combination of dry Spirulina, egg yolk, Dicrateria sp., edible yeast and weaning diet for shrimp, C. intestinalis grew up to average 59 mm and matured after 60 d cultivation. This culture process could be repeated using the artificially cultured mature ascidians as material. When the fertilized eggs were maintained under 10, 15, 20, 25 ℃, they hatched within 30 h, 22 h, 16 h and 12 h 50 min respectively experiencing cleavage, blastulation, gastrulation, neurulation, tailbud stage and tadpole stage, The tadpole larvae were characterized as typical but simplified chordates because of their dorsal nerve cord, notochord and primordial brain. After 8-24 h freely swimming, the tadpole larvae settled on the substrates and metamorphosized within 1-2 d into filter feeding sessile juvenile ascidians. In addition, unfertilized eggs were successfully dechorionated in filtered seawater containing 1% Tripsin, 0.25% EDTA at pH of 10.5 within 40 min. After fertilization, the dechorionated eggs developed well and hatched at normal hatching rate. In conclusion, this paper presented feasible methodology for rearing the tadpole larvae of C. intestinalis into sexual maturity under controlled conditions and detailed observations on the embryogenesis of the laboratory cultured ascidians, which will facilitate developmental and genetic research using this model system.

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