Molecular neuro-physiological study of insect diapause mechanism

昆虫滞育机制的分子神经生理学研究

• 批准号: 10640665
• 负责人: SHIMIZU Isamu
• 金额: $ 1.66万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10640665/
• 关键词: Photoperialic receptor; Bombyx mori; brain; opsin; Visual Pigment; subcloning; eye; Boceropsin; DNA; 単眼; PCR

PCR was carried out using cDNA prepared from brain of the silkworm, Bombyx mori with primers by which opsin encoding genes of the compound eyes can be amplified. We got an amplification of a band corresponding about 200 bases, and then subcloned a fragment of 210 bases DNA (70 amino acids) encoding a opsin-like gene. On the base of this DNA sequence, we conducted 3'-5'-Race method to determine all length of ORF region. The cloned DNA sequence had about 1600 bases and the ORF coded 377 amino acids. It was found that the DNA sequence has a high homology with ManipI reported as one of the photoreceptive pigments in the Manduca sexta compound eyes. We named this opsin like protein in the silkworm brain Boceropisn (bombyx cerebral opsin). One peptide corresponding to the carboxyterminal 25 amino acids of Boceropsin was synthesized. And immunocytochemistry revealed that some pairs of cells of the silkworm brain showed potent immunoreactive to the rabbit antisera of the peptide. These immunoreactive cells is suggested to be a pupative photoreceptor involved in the photoperiodic response in the silkworm.

PCR是使用从蚕的脑，Bombyx Mori制备的cDNA，并用底漆进行了底漆，可以将Opsin编码化合物眼睛的基因进行扩增。我们得到了对应于200个碱基的条带的放大，然后将编码OPSIN样基因的210个碱基DNA（70个氨基酸）的片段取代。在此DNA序列的基础上，我们进行了3'-5'-Race方法来确定ORF区域的所有长度。克隆的DNA序列具有约1600个碱基，ORF编码了377个氨基酸。发现DNA序列与Manipi具有很高的同源性，据报道是Manduca Sexta化合物中的感光色素之一。我们在蚕脑纤维素（Bombyx脑蛋白酶）中命名了这种Opsin Like蛋白。合成了一种与羧蛋白的羧基25氨基酸相对应的肽。免疫细胞化学表明，一些蚕脑细胞对肽的兔抗血清表现出有效的免疫反应。这些免疫反应性细胞被认为是粒虫中光周期反应的能力感受器。

项目成果

I. Shimizu: "Global ecosystem and light environments"Photoenvironments and Evolution (Kyoritsu Publication). (2000)

I.清水：《全球生态系统和光环境》Photoenvironments and Evolution（共立出版社）。

T. Iwasa: "G protein alphasubunit genes in octopus photoreceptor cells"Zoological Science. (in press).

T. Iwasa：“章鱼感光细胞中的 G 蛋白 α 亚基基因”《动物学科学》。

清水 勇: "地球生態系と光環境"「光環境と生物の進化」共立出版社. 30-43 (2000)

Isamu Shimizu：“全球生态系统和光环境”“光环境和生物体的进化”Kyoritsu Shuppansha 30-43（2000）。

I. Shimizu: "Boceropsin : opsin like protein expressed in the silkworm brain."Journal of Japanese Society for Chronobiology. 5. 59 (1999)

I. Shimizu：“Boceropsin：在蚕脑中表达的视蛋白样蛋白质。”日本时间生物学会杂志。

T.Iwasa: "G protein alpha submit genes in Octopus photoneceptor cells"Zoological Science. (in press). (2000)

T.Iwasa：“章鱼感光细胞中的 G 蛋白 α 提交基因”《动物学科学》。