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標題: | Rab11在果蠅感覺神經元之研究 The role of Rab11 in Drosophila sensory neurons |
作者: | Chih-Yu Chou 周芝妤 |
指導教授: | 李秀香(Hsiu-Hsiang Lee) |
關鍵字: | 神經修剪,果蠅感覺神經元,樹突修剪, Neuron pruning,Drosophila sensory neuron,Dendrite pruning, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 當果蠅經歷中文變態的過程中,果蠅感覺神經元的樹突修剪作用是一個必要的過程。樹突修剪作用受到嚴格的調控,任何錯誤調控可能會對神經系統帶來嚴重後果。樹突修剪的詳細機制仍然不清楚。我們發現,Rab11這個小G蛋白會參與果蠅感覺神經元的樹突修剪。為了闡明Rab11的GTP酶對於樹突修剪的重要性,我們利用針對GTP酶活性的兩個突變(constitutively active和dominant negative突變),證明GTP酶的活性對果蠅感覺神經元的樹突修剪是重要的。Rab11蛋白被發現在果蠅S2細胞中,以及感覺神經元之中會和捲曲螺旋蛋白Spindle-F (Spn-F)有交互作用。本實驗室先前研究發現,在果蠅感覺神經元的樹突修剪之中,Spn-F會調控Ik2蛋白激酶的訊號傳遞。我們更進一步發現在Spn-F蛋白C-terminus的區域中有一蛋白質結構域對SpnF和Rab11交互作用是重要的。最後,我們在螢光共軛焦顯微鏡下觀察到在近神經元本體的樹突上Rab11小泡的移動,而且我們也藉由在神經中大量表現突變的Rab11,我們觀察到在樹突修剪中,被切斷的樹突位置附近,Rab11的活化對樹突varicosity的形成是必要的。 During metamorphosis, there is an essential process in Drosophila Class IV dendritic arborization (C4da) sensory neurons called dendrite pruning. Pruning is a tightly controlled process and any mis-regulation could bring serious consequences to the nervous system. The detailed mechanism of pruning is still remains unclear. We identify a small G protein Rab11 which is involved in the dendrite pruning of Drosophila class IV da neuron. To examine whether the GTPase activity of Rab11 is required for dendrite pruning, we generated constitutively active (CA) and dominant negative (DN) form of Rab11. We found that overexpression of CA-Rab11 in class IV da neurons does not affect dendrite pruning, but overexpression of DN-Rab11 impaired dendrite pruning, suggesting the GTPase activity of Rab11 is critical for dendrite pruning of class IV da neuron. Rab11 was detected in complexes with Spindle-F (Spn-F), which mediates Ik2 kinase signaling in class IV da neurons during pruning, not only in S2 cells, but also in neurons. We further identified a region at C-terminus of Spn-F protein that is important for Spn-F to associate with Rab11 in S2 cells. Finally, we characterized the movement of Rab11 vesicles in the proximal dendrites, and found that wild-type and CA-Rab11, but not DN-Rab11, could be observed in varicosities, which occur around the dendrite severing sites during early pruning. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/7723 |
DOI: | 10.6342/NTU201702842 |
全文授權: | 同意授權(全球公開) |
顯示於系所單位: | 分子醫學研究所 |
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ntu-106-1.pdf | 2.77 MB | Adobe PDF | 檢視/開啟 |
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