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Molecular endocrinological study on reproduction in amphioxus

文昌鱼繁殖的分子内分泌学研究

基本信息

批准号：
09640787
负责人：
KUBOKAWA Kaoru
金额：
$ 2.18万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

l. Environmental and ecological study on amphioxus habitat for understanding reproductive phenomena under natural condition(l) Investigations including analysis of growth, division and environmental factors, and observation of habitat by using ROV in Tansei-Maru from August, 1997 to July, 2000.2. Success of breeding and spawning in laboratories and analysis of swimming and reproductive behaviors.(1) Gonadal developments in a female and a male amphioxi bred from autumn in 1999 to summer in 2000 in lab. (2) Success of spawning and fertilization in July in 2000 and growth of larva for three weeks. (3) No correlation between spawning date and environmental conditions except for the requirement of dark condition. (4) A possibility of physiological signal between female and male for starting spawning which is experimented by a loose of a reproductive activity in each sex which kept separately. (5) Different escape reaction in the direction that caused by stimulation of light, spot light, vibration or touch. Touch at tail causes an escape direction to tail, and touch at head causes to either tail or head.3. Hatschek's pit and the gene(l) Discover of the elongation of nerve cord toward Hatscheck's pit which seems to correspond to hypothalamus or median eminence. (2) Possibility of an existence of brain-hypothalamus-pituitary system in amphioxus. (3) cDNA clones from cDNA library constructed from subtraction between Hatsceck's pit and muscle contained approximately 50 kinds of clones which homologous sequences are searched in data base of cDNA sequence. Functional protein concerned with endocrine system areinsulin-like peptide, IGF binding protein, inhibin and so on but mo clones encoded pituitary hormone genes. (4) Expression of obtained clones start by using in situ hybridization on adult and larva.

L.（1）1997年8月至2000年7月在淡水丸进行了文昌鱼生长、分异、环境因子分析和栖息地观察等调查。在实验室中成功繁殖和产卵，并分析游泳和繁殖行为。(1)从1999年秋季到2000年夏季，在实验室饲养的一雌一雄文昌鱼的性腺发育。(2)2000年7月产卵受精成功，仔鱼生长3周。(3)产卵期与环境条件无相关性，但对黑暗条件有要求。(4)雌、雄鱼开始产卵的生理信号的可能性，是通过分开保存的每一种性别的生殖活动的松散来实验的。(5)光刺激、点光刺激、振动刺激、触摸刺激引起的不同方向的逃避反应。触摸尾部会导致尾部的逃逸方向，触摸头部会导致尾部或头部。哈氏窝与基因（1）发现神经索向哈氏窝延伸，似乎与下丘脑或正中隆起相对应。(2)文昌鱼脑-下丘脑-垂体系统存在的可能性。(3)从Hatsceck's pit与肌肉消减文库中获得的cDNA克隆约有50种，在cDNA序列数据库中检索同源序列。与内分泌系统有关的功能蛋白有胰岛素样肽、IGF结合蛋白、胰高血糖素等，但编码垂体激素基因的克隆较少。(4)获得的克隆的表达通过使用成虫和幼虫的原位杂交开始。