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作者(中文):朱盛均
作者(外文):Chu, Sheng-Chun
論文名稱(中文):人類Tau蛋白的微管結合序列在Tau蛋白帕金森氏症中扮演重要的角色
論文名稱(外文):The microtubule binding repeat of human Tau is critical for Tau-mediated PD in Drosophila using a reporter of VMAT-pHluorin.
指導教授(中文):張慧雲
指導教授(外文):Chang, Hui-Yun
口試委員(中文):桑自剛
范聖興
口試委員(外文):Sang, Tzu-Kang
Fan, Seng-Sheen
學位類別:碩士
校院名稱:國立清華大學
系所名稱:系統神經科學研究所
學號:9980554
出版年(民國):102
畢業學年度:101
語文別:英文中文
論文頁數:37
中文關鍵詞:Tau蛋白帕金森氏症
外文關鍵詞:Tau proteinParkinson's diseaseVMATMAPTmicrotubule binding repeat
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帕金森氏症是僅次於阿茲海默症的神經退化性疾病。帕金森氏症的主要症狀起因於基底核中黑質緻密部多巴胺神經的死亡。Tau蛋白(MAPT)是一種和神經退化性疾病相關的微管連接蛋白,也是帕金森氏症中neurofibrillary tangles (NFT)的主要成分。 在此研究中,我們測試Tau蛋白的微管連接序列和Tau蛋白致病有密切的關聯之假說,所以我們突變了全部或部分人類Tau蛋白的微管連接序列,藉由GAL4-UAS系統表現在果蠅的多巴胺神經元,利用VMAT-pHluorin來作為觀測標記。我們發現突變掉四個微管連接序列的確可以減少過度表現野生型Tau蛋白對多巴胺神經元所造成的損傷。這說明了Tau蛋白的微管連接序列在Tau蛋白調控的帕金森氏症中扮演重要的角色。
Parkinson’s disease is the second most common neurodegenerative disease after Alzheimer’s disease. The primary symptoms of Parkinson's disease (PD) result from reduced activity of dopamine-secreting neurons caused by cell death in the pars compacta region of the substantia nigra. Microtubule associated protein Tau (MAPT) is a microtubule-associated protein linked with neurodegenerative diseases. Tau is a major component of the neurofibrillary tangles (NFT) found in PD. To test whether its microtubule binding repeat of Tau may be essential for its pathogenesis, we mutated the four microtubule binding motifs of human Tau and expressed these mutant protein in the dopaminergic neurons of Drosophila using GAL4-UAS system and using VMAT-pHluorin as a reporter. We found that Tau mutation on these four microtubule binding repeat reduced its toxicity compared to the neuronal degeneration of DA neurons induced by wild-type Tau. In addition, this mutant also showed normal VMAT-pHluorin expression pattern when flies aged to 4-week-old. Interestingly, we found FTDP-17 associated mutants effects on DA neuron with different severity, TauG272V>Tau WT>TauR406W. Moreover, Tau G272V blocking VMAT-pHluorin signaling, whereas Tau 406W did not. Taken together we suggested that the microtubule binding repeat of Tau play a critical role for Tau-mediated PD. Inhibition of the interplay of Tau and mitochondria may serve as a therapeutic target.
Abstract ....................................................................................................................... 2
摘要.............................................................................................................................. 3
Introduction ................................................................................................................. 5
Materials & Methods ....................................................................................................8
Results ....................................................................................................................... 11
1. Patterns of TH neurons and VMAT-pHluorin.......................................................... 11
2. Overexpress htauG272V and htauR406W in Drosophila brain .............................. 12
3. Overexpress htauR1-R4 and htauR2,R4 in Drosophila brain .................................. 14
4. Other phenotype caused by overexpress various htau ......................................... 15
Conclusion and discussion.......................................................................................... 17
Figures........................................................................................................................ 21
Figure 1. TH-GAL4 neurons in Drosophila brai........................................................... 21
Figure 2. htau constructs used in this study............................................................... 22
Figure 3. The pattern of PPL2 and PPM3 clusters under htauG272V and htauR406W expression. ................................................................................................................. 23
Figure 4. Quantification of the neuron number of PPL2 and PPM3 clusters under htauG272V and htauR406W expression .....................................................................24
Figure 5. The pattern of PPL2 and PPM3 clusters under htauR1-R4 and htauR2,R4 expression ..................................................................................................................25
Figure 6. Quantification of the neuron number of PPL2 and PPM3 clusters under htauR1-R4 and htauR2,R4 expression. .......................................................................26
Figure 7. The principle and pattern of VMAT-pHluorin. .............................................27
Figure 8A. Patterns of VMAT-pHluorin (green) and immunostaining of TH neurons (blue) under hTauG272V and hTauR406W expression. ..............................................28
Figure 8B. Patterns of VMAT-pHluorin (green) and immunostaining of TH neurons (blue) under hTauR1-R4 and hTauR2,R4 expression. ..................................................29
Figure 9. Other phenotype under tau protein overexpression. ................................. 30
Figure 10. Future work................................................................................................34
Acknowledgements.....................................................................................................35
References ..................................................................................................................36
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