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研究生： 李宣萱
Lee, Hsuan-Hsuan
論文名稱： 長的非編碼核糖核酸AABR07056156.1 (Lnc156) 促進皮質神經細胞的再生
Long non-coding RNA AABR07056156.1 (Lnc156) promotes regeneration of cortical neurons
指導教授： 陳令儀
Chen, Linyi
口試委員: 張壯榮
Chang, Chuang-Rung
黄兆祺
Hwang, Eric
學位類別： 碩士
Master
系所名稱： 生命科學暨醫學院 - 分子醫學研究所
Institute of Molecular Medicine
論文出版年： 2018
畢業學年度： 106
語文別： 中文
論文頁數： 51
中文關鍵詞： 再生 、長的非編碼核醣核酸 、中樞神經 、皮質神經細胞
外文關鍵詞： central nervous system, cortical neurons, HES5
相關次數： 點閱：2  下載：0 
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    • 在哺乳動物中，中樞神經系統損傷後無法自發地再生，是由於抑制性的外在環境以及隨著神經細胞發育而衰退的內在生長能力。因此，促進受損中樞神經細胞再生仍然是一大挑戰。透過了解神經再生過程的機制，有助於提供中樞神經再生的臨床治療。在本篇研究中，我們使用18天大的胚胎大鼠皮質神經細胞作為我們的研究細胞模式。我們發現在皮質神經細胞受傷後的再生期間， lncRNA－AABR07056156.1（簡稱Lnc156）表現量增加。Lnc156 的大量表現不僅顯著增進神經軸突生長，而且明顯促進神經再生； knockdown Lnc156 則會抑制受傷的神經軸突再生。為了進一步研究 Lnc156 在神經再生過程中的調控機制，我們預測並分析 Lnc156 與 miRNA 或蛋白質或 mRNA 之間相互作用的潛力。我們發現，HES5 mRNA 有較高的可能性與 Lnc156 轉錄物相互作用。除此之外，Lnc156 的大量表達降低了 HES5 的表現量，而目前已知 HES5 會抑制神經軸突的生長。綜合我們的結果，由於 Lnc156 在受損的皮質神經細胞再生過程中表現量上升，可能表現量增加使Lnc156 轉錄物與HES5 mRNA 結合，導致 HES5 mRNA 被降解進而促進神經的再生。

    　In mammalian system, the central nervous system (CNS) does not spontaneously regenerate after injury due to the inhibitory extrinsic microenvironment as well as developmentally declined intrinsic growth ability of neurons. Thus, overcoming the limited regeneration capacity of CNS neurons remains challenging. Better understanding of the mechanisms underlying neuronal regeneration should provide insight into the potential treatment for neuronal injury. In this study, we used cortical neurons from embryonic day 18 (E18) rat as our models and found that the expression level of a long non-coding RNA (lncRNA), AABR07056156.1 (Lnc156) was increased during regeneration. Overexpression of Lnc156 not only significantly increases neurite outgrowth but also promotes neuronal regeneration. On the other hand, knockdown of Lnc156 inhibits neurite re-growth. To further clarify the regulatory role of Lnc156 during regeneration, we predicted and analyzed the potential interaction between Lnc156 and miRNA/protein/mRNA. Our results indicate that HES5 mRNA is likely to interact with Lnc156 transcript. Moreover, overexpression of Lnc156 decreased the expression level of HES5. HES5 is known to play an inhibitory role on neurite outgrowth. Together, these findings showed that the expression of Lnc156 was increased during regeneration of injured cortical neurons and Lnc156 transcript may interact with HES5 mRNA to reduce the level of HES5 and thus promote neuronal regeneration.

    摘要.............................................................i 致謝.............................................................iii 目錄.............................................................vii 縮寫清單.........................................................ix 介紹.............................................................1 神經系統的再生過程................................................1 長鏈的非編碼核醣核酸（Long non-coding RNA , LncRNA）...............4 LncRNA對於中樞神經系統的影響.......................................7 Hairy and Enhancer of split 5（HES5）.............................9 材料與方法........................................................10 結果..............................................................16 在大腦皮質神經再生過程中，AABR07056156.1（Lnc156）表現量上升.........16 Lnc156 是一個 lncRNA..............................................17 Lnc156的大量表現增進皮質神經細胞的神經軸突生長.......................17 Lnc156的大量表現促進受傷的皮質神經細胞再生...........................18 Knockdown Lnc156 抑制受傷的皮質神經細胞再生.........................18 Lnc156 和 miRNA 相互作用的可能性...................................19 Lnc156 和蛋白質相互作用的可能性.....................................20 Lnc156 和 RNA 相互作用的可能性.....................................21 討論..............................................................22 參考文獻...........................................................24 附圖..............................................................31 圖一、透過 ChIP-seq 和 RNA-seq 鑑定出在大腦皮質神經再生期間表現量有差異的三個 lncRNA.......................................................31 圖二、在皮質神經細胞再生期間三個lncRNA的表現量.......................32 圖三、Lnc156不具有蛋白質編碼的能力..................................34 圖四、Lnc156的大量表現會增加皮質神經細胞的神經軸突長度和末端端點數目...36 圖五、Lnc156的大量表現促進受傷的皮質神經細胞的神經軸突再生............38 圖六、Knockdown Lnc156會抑制受傷的皮質神經細胞再生...................41 圖七、預測可能與 Lnc156 結合的miRNA.................................43 圖八、預測可能與 Lnc156 轉錄物結合的蛋白質...........................46 圖九、預測可能與 Lnc156 轉錄物結合RNA...............................48 圖十、推測的 Lnc156 作用機制........................................49 表格..............................................................50 Table 1. Primers sequences for miRNA RT-PCR and Q-PCR.............50 Table 2. Primers sequences for PCR and Q-PCR......................51 Table 3. SiRNA oligo duplexes for Lnc156 knockdown................51  

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