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Evolutional Study of Photo-Reception and Photo-Signal Transduction Systems of Animal.

动物光接收和光信号转导系统的进化研究。

基本信息

批准号：
09833005
负责人：
IWASA Tatsuo
金额：
$ 1.73万
依托单位：
Himeji Institute of Technology
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 1998
项目状态：
已结题

项目摘要

In order to clearify the evoulutional relationship among the photoreception systems of the animal, we investigated the photoreceptor pigment and the photo-signaltransducing protein of several dififerent species, especially lower invertebrate such as flatworm and jellyfish. The goat of this research project is to understand the evolutional pathway of the visual system of animals. The results were as follows :1) The full length opsin gene of jellyfish was cloned and sequenced. The anti-serum against the C-teminal synthetic peptides of the deduced sequence was obtained from mice. It was confirmed by northern blot analysis and immuno-histochmical studies with light and electron microscope that the gene was exclusively expressed in the eye.2) From the planariarn eye, Gq and Go class G protein alpha subunit gene were cloned. Insitu hybridization analysis revealed that Gq class of G protein was highly expressed in the planarian eye. The results were also confirmed by the immunohistochemical studies with the Gq-specific antiserum. Based on the above results, it can be safely said that in the planarian visual system rhodopsin couples with Gq class G protein, indicating that the coupling between rhodopsin and Gq was very old.3) The opsin gene was cloned from the brain and ocellus (stemma) of Bombyx larva. The nucleotide sequence was the same in both cDNA clones from brain and that from ocellus, indicating that the same opsin protein was used in the both organs. The antiserum against the C-terminal peptide synthesized according to the nucleotide sequence of the cDNA clone crossreacted with the ocellus of Bombyx larva.

为了阐明动物光感受系统之间的进化关系，我们对几种不同种类的光感受器色素和光信号转导蛋白进行了研究，特别是低等无脊椎动物如扁形虫和水母。本研究的目的是了解动物视觉系统的进化途径。主要研究结果如下：1）克隆了海蜇视蛋白基因的全长序列。从小鼠中获得了针对推导序列的C-肽合成肽的抗血清。通过北方杂交分析和免疫组织化学光镜及电镜观察，证实该基因在真涡虫眼中特异表达。2）从真涡虫眼中克隆了Gq和Go类G蛋白α亚基基因。原位杂交分析显示，Gq类G蛋白在真涡虫眼中高度表达。结果也证实了免疫组化研究与Gq特异性抗血清。根据以上结果，可以肯定地说，在真涡虫视觉系统中，视紫红质与Gq类G蛋白偶联，说明视紫红质与Gq类G蛋白偶联的历史非常悠久。3）从真涡虫幼虫的脑和单眼（干细胞）中克隆了视蛋白基因。结果表明，脑和单眼的视蛋白cDNA克隆的核苷酸序列完全相同，表明脑和单眼的视蛋白均含有相同的视蛋白。根据该cDNA克隆的核苷酸序列合成的C端肽的抗血清可与胭脂虫幼虫的单眼发生交叉反应。