LRRK2是一種調控多種機制的多功能性蛋白，許多研究指出LRRK2突變造成激酶活性上升與帕金森氏症有密切的關係。Tau蛋白不正常的磷酸化會早成tau蛋白堆積在細胞質中，種種不正常的堆積是造成神經細胞凋亡的主要原因之一。近年來，研究指出退化性神經疾病不只影響神經元本體更影響了神經突觸傳遞的功能。在這篇研究中，我們使用果蠅作為模式生物去觀察LRRK2和tau蛋白表現在不同型態的神經元中會對神經突觸的傳遞造成甚麼影響。我們發現在多巴胺神經細胞上，tau蛋白的過量表現會抑制神經突觸傳遞，但同時表現LRRK2時則可以恢復神經突觸的傳遞，然而，同時表現突變型的LRRK2G2019S時則會更加減弱神經突觸間的傳遞。在谷氨酸神經細胞上，過量表現tau蛋白一樣會降低神經突觸的傳遞，但同時表現LRRK2及LRRK2G2019S則不會對神經突觸間的傳遞有更進一步的影響。總結來說，LRRK2在不同型態的神經細胞中可能會造成不同的效果。其他研究也指出在神經退化性疾病患者的腦組織中有較高的氧化壓力。較高的氧化壓力是造成神經細胞凋亡的原因之一，體內負責調節氧化壓力的抗氧化壓力蛋白就顯得非常重要。Nrf2是一種能夠藉由與ARE片段結合去激活下游抗氧化壓力蛋白生成的一種轉錄因子。我們發現，無論在多巴胺神經細胞還是谷氨酸神經細胞中過量表現tau蛋白都會降低Nrf2的表現量，而同時表達LRRK2則可以回復Nrf2的表現量。此外，在多巴胺神經中表達突變LRRK2G2019S時則會使Nrf2的表現量變得更少。總而言之，我們的研究突顯了LRRK2和tau蛋白在許多方面都有功能性上互相影響的可能性。

LRRK2 is a multi-functional protein regulating many mechanisms. Many studies have reported that LRRK2 mutation with higher kinase activity is highly related to Parkinson’s disease. Expression of tau causes tau protein aggregate in the cytosol, which leads to neuron death in many neurodegenerative diseases. Recently, many studies show that not only neurons but synaptic function is affected in the patient’s brain. In this study, we use Drosophila as a model to investigate how LRRK2 and tau affect neurotransmission on the different types of neurons in the brain. We find that overexpression of Tau in the dopaminergic system disrupted neurotransmission at the mushroom body but co-expression of LRRK2 can rescue this defect. However, expression of LRRK2G2019S exacerbated the deficit. Although expression of tau in the glutamatergic neurons caused a reduction of the neurotransmission at antenna mechanosensory and motor center, expression of LRRK2 does not rescue the neurotransmission. Moreover, LRRK2G2019S does not worsen the defect. Taken together, LRRK2 may have different effect in different neuron systems. Studies also indicated that higher oxidative stress occurs in neurodegenerative disease patient’s brain and oxidative stress may induce neuron death in the brain. Antioxidant proteins are important to deal with oxidative stress. It is known that Nrf2 can bind to ARE and activate downstream antioxidant proteins synthesis. In present study, we find that overexpression of tau in the dopaminergic or the glutamatergic neurons could reduce Nrf2 level in the brain but expression of LRRK2 can rescue the defect. In addition, expression of LRRK2G2019S in the dopaminergic neurons aggravates defect. In summary, our study shows LRRK2 and tau have functional interaction in many aspects.  

Table of Contents
Abstract I
摘要 II
Chapter 1. Introduction 8
1.1 Introduction of Parkinson’s disease 8
1.1.2 Symptoms of Parkinson’s disease 8
1.1.2.1 Motor symptoms 9
1.1.2.2 Non-motor symptoms 10
1.1.3 Pathology of Parkinson’s disease 11
1.2 Introduction of Leucine-rich repeat kinase 2（LRRK2) 11
1.2.1 LRRK2 and synaptic function 11
1.2.2 LRRK2 and reactive oxygen species 12
1.2.3 LRRK2 mutations 13
1.3 Introduction of microtubule-associated protein Tau 13
1.3.1 Tau protein and synaptic function 14
1.3.2 Tau phosphorylation 14
1.3.3 Kinase of Tau protein 15
1.4. Introduction of nuclear factor E2-like 2 16
1.4.1 Physiology of Nrf2 16
1.4.2 Nrf2 and oxidative stress 17
1.5 Introduction of drosophila 18
1.6 Motivation and research outline 19
Chapter 2. Materials and Methods 20
2.1 Drosophila strain and culture 20
2.2 Adult brain dissection 20
2.3 Life span assay 20
2.4 Locomotor assay 21
Chapter 3. Results 22
3.1 Overexpression of LRRK2 and LRRK2G2109S in the dopaminergic neurons and the glutamatergic neurons 22
3.2 Overexpression of human wild type tau_1 and wild type tau_4 in the dopaminergic neurons and the glutamatergic neurons. 24
3.3 Expression of LRRK2 rescues neurotransmission disruption caused by human wild type tau_1 and tau_4. 25
3.4 Expression of LRRK2G2019S not exacerbate neurotransmission deficit caused by wild type tau_4. 27
3.5 Expression of LRRK2 and LRRK2G2019S affect Nrf2 level in fly’s brain. 28
3.6 Expression of human wild type tau_1 and wild type tau_4 reduce Nrf2 level in the fly’s brain in 7th week. 29
3.7 Expression of LRRK2 rescues decreased Nrf2 level caused by human wild type tau_1 and tau_4. 29
3.8 Expression of LRRK2G2019S exacerbates decreased Nrf2 level caused by human tau_4. 30
3.9 The life spans of fly over-express LRRK2, LRRK2G2019S, human wild type tau_1, and human wild type tau_4 individually in the dopaminergic and the glutamatergic neurons. 31
3.10 The life spans of fly co-express LRRK2 or LRRK2G2019S with human wild type tau_1 and human wild type tau_4 in the dopaminergic and the glutamatergic neurons. 33
3.11 The locomotor ability of flies expressed different genes in the dopaminergic neurons at 1-week, 4-week and 7-week. 34
3.12 The locomotor ability of flies expressed different genes in the glutamatergic neurons at 1-week, 4-week and 7-week. 35
Chapter 4. Discussion 37
4.1 LRRK2 shows a protective effect on neurotransmission. 37
4.2 LRRK2 protects Nrf2 from degradation 40
Chapter 5. Figures 42
Chapter 6. References 79

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