Evolutional study on the endocrine system of brain-pituitary interaction in amphioxus

1.对文昌鱼(Branchiostoma Belcheri)Hatschek窝和神经管的cDNA文库进行了EST分析，为脑-垂体内分泌系统的进化研究奠定了基础。目前的研究结果如下：(1)成功去除神经管包括脑部，使构建的文库获得高质量的文库。神经管的EST项目从2002年开始，作为与教授的合作项目。K.Agata和S.Kuratani，Riken，Kobe。对3800个无冗余克隆的核苷酸序列进行了分析，直到2002年3月。该项目继续从文昌鱼神经管中获得所有的cDNA克隆。获得了甲状旁腺激素、RFamid、降钙素和多种生长因子等激素基因。在不久的将来，文昌鱼的EST结果将通过与文昌鱼神经管和脊椎动物脑中存在的特定基因的分子结构和生理机制的比较，揭示脑从神经管进化的过程。(2)从哈茨坑中获得的cDNA文库包括来自这个小坑的肌肉、皮肤和肾脏。PIT中可能具有特异性的DNA序列有胰岛素样生长因子结合蛋白、胰岛素样蛋白、铁蛋白、淀粉样蛋白、钙相关蛋白等。对文昌鱼的这些基因进行了系统发育研究。(3)文昌鱼的这些结果为研究脊椎动物和哺乳动物脑内神经递质的作用机制和相互作用奠定了基础。文昌鱼成体原位杂交方法的建立。日本文昌鱼实验室自然产卵首次成功。在繁殖期2个月内，通过电影对其产卵行为进行调查，确定产卵条件，估计影响产卵成功的行为和环境因素。

1. EST analysis of cDNA libraries from Hatschek's pit and neural tube in amphioxus (Branchiostoma belcheri) was attempted in order to the evolutional study on brain-pituitary endocrine system. The results so far are as follows.(1) The successfully removal of he neural tube included brain part enabled the high quality of constructed cDNA library. The EST project of neural tube has been started from 2002 as the collaboration with Profs. K. Agata and S. Kuratani, Riken, Kobe. Nucleotide sequences of the 3800 clones without redundant was analyzed until March 2002. The project continues to obtain all of cDNA clones from amphioxus neural tube. A few hormone genes were obtained such as parathyroid hormone, RFamid, calcitonin, and some kinds of growth factors. In near future, the EST results in amphioxus will reveal an evolution of brain from the neural tube by the comparison of molecule structure and physiological mechanism with specific genes which exist both in amphioxus neural tube and vertebrate brain.(2) The cDNA library from Hatschek's pit includes muscle, skin and nephridium from that the small size of the pit. DNA sequences which might be specific in the pit was Insulin like growth factor binding protein, Insulin like protein, Ferritin, Amyloid, Calcium related proteins. These genes in amphioxus were phylogenetically studied. (3) These results generated from amphioxus contribute the study on mechanism and interactions of neurotransmitters in the brain of vertebrate and mammal.2. Establishment of in situ hybridization in adult amphioxus.3. First success of natural spawning of amphioxus in the laboratory in Japan. Spawning behavior was investigated by movie during reproductive period, two months, and determined the spawning conditions and estimated behavioral and environmental factors for success of spawning.