2006-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/691034摘要：在昆蟲發育與生殖的過程中 neuropeptides 扮演關鍵性的角色。在眾多已純化的 neuropeptides中，corazonin 已證實會造成蜚蠊與蛾類的心跳加速，以及引起飛蝗的行為及體色變化。最近在蛾類的研究發現 corazonin 基因產物與時鐘基因 period 產物出現在腦中的相同細胞中。因此我們推測corazonin對於昆蟲的日週律動有影響力。為了證實這個假說，首先我們將選殖 corazonin 基因以得到其 cDNA 序列資料，接著以此資料可以進一步知道 corazonin 基因的 mRNA 與氨基酸序列的資料，據此可以進行原位雜合與免疫染色的工作，以證實德國蜚蠊的 corazonin 基因與時鐘基因 period 是否在腦中的相同細胞表現。然後注射 corazonin 至德國蜚蠊的體內，再記錄其對德國蜚蠊的活動行為日週律動、蛻皮激素的濃度及電生理活性的影響。接著我們將同樣選殖了corazonin 的受器基因，進而以免疫染色的方法找到 corazonin 受器的位置，據此得知 corazonin 作用在德國蜚蠊體內的標的位置；此外，本計畫也將針對另一種 neurop<br> Abstract: The neuropeptides play a key role in the process of development and reproduction in insects. A neuropeptide, Corazonin was shown to have diversed functions in various insects (heart beat in cockroach and moth; changes of behavior and body coloration in Locust). Recently, the period and corazonin genes were found to be colocalized within the same cells in a moth (Galleria mellonella). We hypothesize that corazonin may be able to exert influence on the circadian rhythm in insects. To test the hypothesis, the cDNA of corazonin gene will be cloned from the German cockroach (Blattella germanica) followed by sequence analysis, in situ hybridization and immunostaining to shown the co-localization of Corazonin and PER. Next, Corazonin will be injected into the German cockroach. The locomotor circadian rhythm, concentration of ecdysone and electrophysiological activity will then be recorded to understand the effects of Corazonin. Thirdly, corazonin receptor gene will be cloned in order to know the target sites of Corazonin in the German cockroach. Moreover, we will also use another neuropeptide, the pigment dispersing factor (PDF), a factor that has been demonstrated to influence the locomotor circadian rhythm, to explore the unknown mechanisms and pathways underlying the expression of circadian rhythm. In this project, we will in addition clone the PDF receptor to understand the distribution of target cells of PDF. By investigating both corazonin and PDF, we will understand the possible mechanisms and pathways underlying the circadian rhythm expression in the German cockroach.德國蜚蠊日週律動色素分散因子German cockroachcircadian rhythmpigment dispersing factorcorazonin.