臺灣博碩士論文加值系統

unpaired (upd)的蛋白質產物為Jak/STAT訊號傳遞鏈的配體(ligand)。upd表現於眼碟後緣的中心點，亦即型態溝(morphogenetic furrow, MF)起始的位置。因此upd基因可能與型態溝的起始有關。在之前的研究中已經知道N、Dpp、Hh、Wg等訊號會參與在型態溝的起始機轉中，但upd基因的調控機轉仍然未知。為明白如何調控upd基因的表現，我們探討這幾個訊號傳遞鏈與upd基因的交互作用。經由功能獲得(gain-of-function)與功能喪失(loss-of-function)的實驗發現：Hh、Dpp訊息必須和N合作才能活化upd的表現。此外我們也發現Wg訊息會抑制upd的表現，並且在Jak/STAT訊號傳遞鏈中具有自我回饋(auto-regulation)的機轉。我們對於這些訊息傳遞鏈是否直接調控upd基因表現也相當感興趣。藉由找尋增強子(enhancer)，並在增強子上面所具有的轉錄因子結合點(binding site)做點突變，藉由這些實驗來確認會直接調控upd表現的訊息傳遞鏈。我們發現N、Hh和Wg都可能直接透過我們找到的增強子來調控upd基因的表現。
在本篇研究中，我們證明了upd基因在空間與時間上所受到的調控。N、Dpp與Hh訊息表現的交叉點決定了upd基因表現的位置。我們也發現Wg訊息會抑制upd使upd基因只在眼碟後緣中央表現，而Jak/STAT訊息傳遞鏈可能透過自我回饋機轉維持upd基因的表現。

unpaired (upd) encodes a secreted ligand for the Jak/STAT pathway. It is expressed at the central point of the posterior margin in the eye disc, the initiation site of the morphogenetic furrow (MF). Thus upd may play the role in MF determination. It had been shown that several signaling pathways are involved in MF initiation, such as N, Dpp, Hh and Wg. But it is still unclear what factors specify the expression of upd. To understand the regulatory network controlling upd expression, we checked the interactions between upd and several signaling pathways. Testing of gain-of-function and loss-of-function effects of these pathways on the expression of upd-lacZ suggested Hh and Dpp pathway cooperate with N signaling to activate upd expression at the posterior center of the eye disc. Furthermore, we found that Wg signaling may negatively regulate upd expression, and Upd was also involved in auto-regulation of its own expression. We are also interesting in finding the direct regulation of upd. We checked the direct regulation by enhancer identifications and binding site mutations of several transcriptional factors. We found N, Hh and Wg signaling may regulate upd through these enhancers.
In this study, we demonstrated the spatial and temporal control of upd expression. The intersection of N and Dpp, Hh signaling determines the upd expression. We also discovered a novel finding that Wg signaling restricts the upd expression in posterior center and Jak/STAT pathway may have auto-regulation to maintain upd expression.

中文摘要 2
Abstract 3
Introduction 4
Result 10
Discussion 16
Materials and Methods 21
Reference 25
Figures and Table 33

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