Studies on investigation and distribution of the neuropeptides in crustaceans

甲壳类神经肽的调查与分布研究

• 批准号: 13660209
• 负责人: YASUDA Akikazu
• 金额: $ 2.37万
• 依托单位: Suntory Institute for Bioorganic Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660209/
• 关键词: red swamp crayfish; MALDI-TOF MS; neuropeptide; laser microdissection; orcokinin; SIFamide; tachykinin-related peptide; accessory lobe; Gタンパク質共役型受容体; 肝膵臓

Matrix-assisted laser desorption ionization with time-of-flight mass spectrometry (MALDI-TOF MS) has become a rapid and sensitive analytical tool that is having an increasing impact on the direct MS measurement of the secretory granules in the cells. The result is available for the second step that is purification of a given material by monitoring its molecular mass and then sequencing it. The third step is molecular cloning for the precursor of a given substance based on the structural information. The final step is capillary reversed-phase HPLC/Q-Tof analysis with the tissue extract to determine the posttranslational modifications for mature hormone molecules. We have called this total strategy as "topological mass spectrometry analysis". In addition, laser microdissection has become a powerful tool for construction of mass spectrometric morphology in the brain since it is possible to collect single cells from a thin tissue section. In this study, we have identified orcokinin-related peptides, crustacean-SIFamide, and decapod tachykinin-related peptide from the crayfish brain. It is known that the olfactory lobe receives the primary afferent endings of the chemoreceptors on antenna I, but the accessory lobe does not appear to receive a direct input from primary afferent axons. The output pathways from the, olfactory lobe and accessory lobe are provided by the axons of a large population of projection neurons, in which projection neurons innervating the olfactory lobe and accessory lobe innervate the neuropiles of the medulla terminalis and the hemiellipsoid body. The peptides seem to play a significant role in olfaction. Finally, topological mass spectrometry analysis will provide excellent means in micro-characterization of protein/peptide hormone in the field of fish. and insect endocrinology.

基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）已成为一种快速，灵敏的分析工具，具有越来越大的影响，在细胞中的分泌颗粒的直接MS测量。结果可用于第二步，即通过监测其分子量来纯化给定物质，然后对其进行测序;第三步是基于结构信息对给定物质的前体进行分子克隆。最后一步是用组织提取物进行毛细管反相HPLC/Q-Tof分析，以确定成熟激素分子的翻译后修饰。我们把这种总体策略称为"拓扑质谱分析"。此外，激光显微切割已成为一个强大的工具，用于构建质谱形态学在大脑中，因为它可以收集单细胞从薄的组织切片。在这项研究中，我们已经确定了orcokinin相关肽，甲壳类SIFamide，十足类速激肽相关肽从螯虾脑。已知嗅叶接收天线I上的化学感受器的初级传入末梢，但副叶似乎不接收来自初级传入轴突的直接输入。嗅叶和副叶的输出通路由大量投射神经元的轴突提供，其中支配嗅叶和副叶的投射神经元支配终髓和半椭圆体的神经桩。这些肽似乎在嗅觉中起着重要作用。最后，拓扑质谱分析将为鱼类蛋白质/肽类激素的微观表征提供良好的手段。和昆虫内分泌学

项目成果

Yasuda-Kamatani, Y., Yasuda, A.: "Cloning and expression of a cDNA for a putative G protein-coupled receptor from the hematopancreas of the crayfish, Procambarus clarkii."Gen.Comp.Endocrinol.. 125. 25-33 (2002)

Yasuda-Kamatani, Y., Yasuda, A.：“来自克氏原螯虾血胰腺的推定 G 蛋白偶联受体 cDNA 的克隆和表达。”Gen.Comp.Endocrinol.. 125. 25-33

Yasuda, A.: "Capillary LC-MS analysis of the protein/peptide hormones (in Japanese)"Newsletter of JSCE. 107. 29-35 (2002)

Yasuda, A.：“蛋白质/肽激素的毛细管 LC-MS 分析（日语）”JSCE 时事通讯。

Yasuda-Kamatani, Y., Yasuda.A.: "Cloning and expression of a cDNA for putative G protein-coupled from receptor the hepatopancreas of the crayfish. Procambarus clarkii"General and Comparative Endocrinology. 125. 25-33 (2002)

Yasuda-Kamatani, Y., Yasuda.A.：“克隆和表达与小龙虾肝胰腺受体偶联的推定 G 蛋白 cDNA。克氏原螯虾”普通和比较内分泌学。

Yasuda, A., Yasuda-Kamatani, Y., Nakajima, T.: "Perspective in Comparative Endocrinology"A new strategy for exploring hormones by topological mass spectrometry analysis.. 6 (2001)

安田，A.，安田-镰谷，Y.，中岛，T.：“比较内分泌学的视角”通过拓扑质谱分析探索激素的新策略.. 6 (2001)

Takahashi, A., Amemiya, Y., Yasuda, A., Meguro, H., Kawauchi, H.: "Mass spectrometric detection of proopiomelanocortin (POMC)-related peptides following molecular cloning of POMC cDNA in bigeye tunna Thunnus obesus."Fisheries Science. 68. 1073-1080 (2002)

Takahashi, A.、Amemiya, Y.、Yasuda, A.、Meguro, H.、Kawauchi, H.：“在大眼金枪鱼 Thunnus obesus 中对 POMC cDNA 进行分子克隆后，对阿片黑皮素原 (POMC) 相关肽进行质谱检测。”