摘要


磷脂質PI(3,4,5)P3在PI3K細胞訊息傳遞路徑中是扮演第二傳訊者(second messenger)的角色。胰島素(insulin)刺激血管內皮細胞產生一氧化氮(nitric oxide)進而使血管舒張及增加血流量的調控機制也涉及PI3K細胞訊息傳遞路徑。然而一氧化氮也曾經被報導過產生在神經細胞裡面，在此一氧化氮的作用是神經傳導物質(neurotransmitter)。此外，一氧化氮也被報導在孤立束核(NTS)的中樞血壓調控裡扮演著重要的角色。先前在我們實驗室裡，將胰島素微注射到大鼠的孤立束核裡會造成明顯的血壓降低以及心博舒緩的現象，另外也會造成PI3K下游的蛋白激酶Akt的活化。為了研究胰島素在孤立束核裡造成一氧化氮產生更詳細的下游訊息傳遞機制，我們在神經細胞株PC12和GH3裡以及在大鼠的孤立束核裡，研究脂類第二傳信者- PI(3,4,5)P3對一氧化氮產生的影響。將PC12和GH3細胞以10

Abstract


The phospholipid PI(3,4,5)P3 works as a second messenger in PI3K signaling pathway. The PI3K signaling pathway is involved in insulin stimulated nitric oxide (NO) production in vascular endothelium, leading to vasodilation and increased blood flow. However, the production of NO also has been reported in neurons as a neurotransmitter and in nucleus tractus solitarii (NTS), NO plays a role in central cardiovascular regulation. Previously, microinjection of insulin into the NTS of rats produces prominent depressor and bradycardic and activates the PI3K downstream Akt. Therefore, to investigate the detail downstream signaling of insulin stimulated NO production in NTS, the effects of PI(3,4,5)P3 on NO production were determined in neuronal cell lines PC12 and GH3 and in NTS of SD rats. The GH3 and differentiated PC12 exposed to 10

Contents


Contents 1
Abbreviations 2
中文摘要 4
English Abstract 6
Chapter 1
General Introduction
1.1 Backgrounds and Significance 9
1.2 Specific Aims 20
1.3 References 21
Chapter 2
Lipid Second Messenger Activated Akt/PKB Signaling in the Nucleus Tractus Solitarii Contributes to Central Cardiovascular Regulation
2.1 Summary 28
2.2 Introduction 30
2.3 Materials and Methods 34
2.4 Results 41
2.5 Discussion 47
2.6 References 54
Figures and Figure Legends 59
Chapter 3
Roles of Neurogranin and Neuromodulin in Nucleus Tractus Solitarii
3.1 Summary 84
3.2 Introduction 85
3.3 Materials and Methods 89
3.4 Results 101
3.5 Discussion 106
3.6 References 110
Figures and Figure Legends 114
Chapter 4
Future Perspectives
4.1 Future Perspectives 134
4.2 References 138
Appendix 139

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