蚵岩螺(Thais clavigera)為亞太地區潮間帶常見腹足類生物，過去已證實會受有機錫影響而造成雄化(imposex)，常用於評估有機錫汙染。但近年臺灣西海岸部分地區的雄化現象有所趨緩，雌螺數量約為雄螺之2倍，且可發現雄性陰莖較短小之情形；然而，此現象無法單以形態特徵加以分析解釋。過去水生生物研究中，類雌性環境荷爾蒙的污染常以卵黃原生成作用(vitellogenesis)進行評估；近期有研究將蚵岩螺部分基因定序並辨識，其中RXR (retinoid X receptor)基因屬核受體家族基因，功能為調控生長與細胞分化，且被認為是啟動雄化的關鍵基因；另有雌激素受體基因(estrogen receptor, ER)與卵黃原蛋白基因(vitellogenin, VTG)，這二基因分別參與調控雌性特徵發育與卵黃原生成作用。本研究之目的為建立蚵岩螺三種生殖發育相關基因表現量之偵測方法。透過設計對各基因有專一性的引子，配合即時定量聚合酶連鎖反應(RT-qPCR)方法，調整RT-qPCR反應條件並利用凝膠電泳進行檢測確認引子的專一性與適當之稀釋倍率。本研究建立高靈敏度的即時定量聚合酶(RT-qPCR)反應可偵測蚵岩螺的基因表現，對過去利用雄化探討有機錫汙染會受到不同類型環境荷爾蒙影響而失去指標性的問題加以改善，在環境荷爾蒙影響蚵岩螺的研究上有進一步提升。

Oyster drill (Thais clavigera) is a common intertidal snail in Asia Pacific. It is a widely used bioindicator for monitoring organotin pollution shown as female snail with imposex. However, female skewed sex ratio and reduced penis size in males are observed in Taiwan recently. This phenomenon can’t be explained based on previous morphological and physiological studies. In order to gain better understanding on the abnormality of the oyster drill, the expression of genes in relation to reproduction of aquatic snails including RXR (retinoid X receptor), ER (estrogen receptor) and VTG (vitellogenin) were explored. Therefore, the purpose of this study was to establish a RT-qPCR (real-time quantitative polymerase chain reaction) method to detect the expression level of these three reproductive and development related genes in T. clavigera. By adjusting the reaction conditions and quality checking through gel electrophoresis, the target genes could be amplified specifically. A quantitative method to determine gene expressions of environmental monitoring species T. clavigera was established. Subsequently, variations in different tissues or samples from different sites were evaluated. Moreover, the assessment of the influence of endocrine disruption chemicals on marine snails can be further extended.

論文審定書 i
論文公開授權書 ii
誌謝 iii
中文摘要 iv
英文摘要 v
目錄 vi
表目錄 vii
圖目錄 viii
壹、前言 1
貳、材料方法 5
參、結果 10
肆、討論 15
伍、參考文獻 20

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