Neuropeptide hormone of insect : molecular mechanisms and tissue responses

昆虫神经肽激素：分子机制和组织反应

• 批准号: 06304005
• 负责人: SAKURAI Sho
• 金额: $ 12.35万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06304005/
• 关键词: neuropeptide hormone; bombyxin; prothoracicotropic hormone; diapausing hormone; neurosecretion; 昆虫; ペプチドホルモン; 神経分泌ホルモン; PBAN; 変態; 寄生

The present scientific research project has been carried out over 3 years from the 1996 to 1998 fiscal year. The initial objective was elucidation of the molecular mechanisms underlying insect metamorphosis and embryonic diapause in respect to the expression of neuropeptide hormones and their physiological effects and we consider to have reached the goal at an appreciable level.Bombyxin, an insulin-related neuropeptide, consists of 30 gene copies and their expression pattern in Bombyx genome and the nucleotide sequences of upstream region of all the 30 genes have been accomplished. Bombyxin receptor has also been elucidated by cDNA cloning to possess the tyrosine kinase domein similar to insulin receptor. Ultra-micro assay method of bombyxin was developed using time-resolved fluoroimmunoassay (TR-FIA) which enabled to quantify a very small amount of the hormone as low as few atto-moles and to determine the detailed changes in hemolymph bombyxin concentrations through the larval-pupal-a … More dult period. The same method became applicable to measurement of hemolymph PTTH titer, which showed a daily secretion of PTTH even before head critical period (HCP). PTTH secretion is stimulated by a neurotransmitter, acetylcholine, indicating that acetylcholine neuron may be involved in the regulation of PTTH cells. As a part of elucidation of molecular mechanisms of PTTH stimulation of prothoracic glands, purification of PTTH receptor and elucidation of its physical properties are in progress. Expression dynamics of diapause hormone (DH) mRNA showed to be different in embryonic and larval stages of animals that are destined to produce diapausing eggs from those to produce non-diapausing eggs. One of the major DH effects was involved in the expression of trehalase gene in ovary. DH gene was expressed in 3 clusters of neurosecretory cells in suboesophageal ganglion. DH is processed and secreted from the posterior cluster while pheromone biosynthesis activating neurohormone which is processed from the same precursor molecule as DH is secreted from the anterior and middle clusters. As described above, the present research project succeeded to give fundamental knowledge on understanding the molecular mechanisms of neuropeptide hormones involved in insect metamorphosis. Less

目前的科研项目是在1996 - 1998财政年度的3年多时间里进行的。最初的目的是阐明昆虫变态和胚胎滞育的分子机制，在神经肽激素的表达及其生理作用方面，我们认为已经达到了一个可观的水平。Bombyxin是一种胰岛素相关的神经肽，由30个基因拷贝组成，在家蚕基因组中的表达模式已经完成，所有30个基因的上游区域核苷酸序列已经完成。Bombyxin受体也通过cDNA克隆被证实具有类似于胰岛素受体的酪氨酸激酶结构域。采用时间分辨荧光免疫分析法（TR-FIA）建立了蚕蛹素的超微量测定方法，该方法可以定量测定极少量的激素，低至几个atto- mols，并测定了幼虫-蛹-更成年期血淋巴蚕蛹素浓度的详细变化。同样的方法也适用于血淋巴中甲状旁腺激素滴度的测定，结果显示，在头临界期（HCP）之前，甲状旁腺激素的分泌量每天都在增加。甲状旁腺激素的分泌受神经递质乙酰胆碱的刺激，提示乙酰胆碱神经元可能参与了甲状旁腺激素细胞的调控。作为PTTH刺激前胸腺分子机制研究的一部分，PTTH受体的纯化及其物理性质的研究正在进行中。滞育激素（DH） mRNA的表达动态在滞育卵和非滞育卵动物的胚胎期和幼虫期有所不同。其中一个主要的DH效应涉及到海藻酶基因在卵巢中的表达。DH基因在食管下神经节3簇神经分泌细胞中表达。DH由后簇加工和分泌，而信息素生物合成激活神经激素由与DH相同的前体分子加工而成，由前簇和中间簇分泌。如上所述，本研究项目成功地为了解昆虫变态过程中神经肽激素的分子机制提供了基础知识。少

项目成果

Nomura, Y.: "Purification and characterization of hemolymph3-oxoecdysteroid 3b-reductase of the silkworm, Bombyx mori." Insect Biochem.Mol.Biol.26. 249-257 (1996)

Nomura, Y.：“家蚕血淋巴 3-氧化蜕皮类固醇 3b-还原酶的纯化和表征。”

Iwami, M.: "Bombyxin-related peptides : cDNA structure and expression in the brain of the hornworm Agrius convoluvuli." Insect Biochem.Mol.Biol.26. 25-32 (1996)

Iwami, M.：“家蚕素相关肽：cDNA 结构和在角虫 Agrius convoluvuli 大脑中的表达。”

M.Tanaka,et al.: "Morphological changes of BM-N4 cells induced by bombyxin,an insulin-related peptide of Bombyx mori." Regulatory Peptides. (in press).

M.Tanaka 等人：“家蚕素（一种家蚕胰岛素相关肽）诱导的 BM-N4 细胞的形态变化。”

K.Nagata,et al.: "Three-dimensional strucutre and receptor-recognition surface of Bombyxin-II,an insulin-related peptide of the silkmoth." Peptide Chemistry. (in press).

K.Nagata 等人：“蚕蛾胰岛素相关肽 Bombyxin-II 的三维结构和受体识别表面。”

Rybcztbski, R.: "Bombyx and Manduca prothoracicotropic hormones : an immunologic test for relatedness." Gen.Comp.Endocri.102. 247-354 (1996)

Rybcztbski, R.：“家蚕和天蛾促胸腺激素：相关性的免疫学测试。”