2001-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/688155摘要：由於德國蜚蠊活動行為的表現上，雌蟲會因為卵巢的發育，而造成活動行為日週律動被掩蓋的現象，本計劃將嘗試深入探討此掩蓋機制。首先我們將利用本研究室 clone 出來的週期 (period, per) 基因，偵測雌蟲 per mRNA 之日週律動表現來證實德國蜚蠊雌蟲之時間訊息產生的分子機制與雄蟲一致，並且與黃果蠅 (Drosophila melanogaster) 之分子調控機制相似。此試驗亦將揭露卵巢發育所造成的活動行為掩蓋現象是否會波及時間訊號產生之日週律動。另外，我們將進行視網膜電位圖譜 (Electroretinogram, ERG) 的偵測，當作第二個生物時鐘外在表徵：利用單一雌蟲之活動行為日週律動的偵測，配合長時期 ERG 的日週律動表現，我們可以了解雌蟲活動行為之掩蓋現象其作用之機制位於生物時鐘系統的哪一部位。我們亦將利用 clone 出來的 per 基因製造出核酸探針來進行原位雜合反應 (in situ hybridization)，藉以鑑別德國蜚蠊大腦視葉中的時鐘細胞。成功檢測出時鐘細胞之後，可以進一步利用「神經元移除」技術，來判別各個細胞在活動行為日週律動調控之中扮演的<br> Abstract: In the performance of locomotor behavior, the development of overies will cause a masking effect on the locomotor circadian rhythm of the female German cockroach. In this study, we propose to investigate this masking mechanism in detail. Firstly, We are going to monitor the expression of circadian rhythm in period (per) mRNA of female adults by the newly cloned per gene from our laboratory. We hope to prove the same molecular mechanisms involved in time signal generation as found in male German cockroach and fruit fly (Drosophila melanogaster). The result of this experiment also can show whether the masking effect on locomotor circadian rhythm will also interfere with the locomotor circadian rhythm. In addition, we will employ Electroretinogram as the second overt rhythm of the biological clock in the German cockroach. By monitoring the circadian rhythm expression in locomotion and visual sensitivity of females, we can understand the action site of the masking factor in circadian rhythm system. We are going to identify the clock cells in the optic lobe by in situ hybridization with the per gene probe. Once all the clock cells are located, we will determine the role of each clock cell by removing each one in turn. Since the hormonal factor released from developing ovaries cause the masking effect on locomotion, we will use immunostaining or in situ hybridization against ecdysone receptor to reveal the action site. By comparing the ecdysone receptor sites and the circadian rhythm pathways, we will unveil the mechanism of masking effect.掩蓋效應日週律動週期基因視網膜電位圖譜蛻皮激素受器masking effectcircadian rhythmperiod geneElectroretinogramecdysone receptor