2008-02-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/711100摘要：神經系統內衝動的傳導，主要決定於神經傳導物質的釋放與接收，如果神經傳導物質的釋放機制有缺陷，會造成各種不同的神經性疾病。直至今日，對於這些疾病的分子與細胞機制所知甚少，導致這些疾病無法根治。雖然過去研究指出細胞內次級信使 (如Ca2+、cAMP) 對傳導物質釋放過程 (稱為胞吐作用) 至為重要，但這些次級信使的角色及互相之間的交互作用，及其下游訊息傳導的分子機制，尚未完全了解，本研究旨在探討cAMP對於胞吐作用融合孔 (fusion pore) 的調控機制。我們將結合分子生物學、細胞培養、電生理記錄及活細胞影像分析技術，探討下列三個主要問題：(1) cAMP是否直接影響胞吐作用的融合孔？或與Ca2+交互作用影響融合孔？(2) cAMP是藉由PKA-dependent或是PKA-independent的機制來影響胞吐作用？(3) cAMP是否對胞吐作用參與的蛋白 (如鈣離子感應蛋白、融合孔組成蛋白，或調控細胞可興奮性的離子通道) 產生影響，進而調控胞吐作用動態的機制？此研究將深入了解次級信使對細胞內胞吐作用的調控，提供神經傳導物質釋放之相關研究全新的方向，對於相關神經性疾病 (如精神分裂症及老年失智症) 提供新的研究視野，期使新的醫療方法的產生。<br> Abstract: Neurotransmission mainly depends on neurotransmitter release and reception. Especially, any defects in transmitter release would lead to various neurological diseases. To date it remains largely unknown about the molecular and cellular mechanisms underlying transmitter release, so no cures have been found for the relevant neurological diseases. Previous studies indicate that cellular second messengers, such as Ca2+ and cAMP, are essential for the process of transmitter release termed as regulated exocytosis. However, it is unknown how these second messengers can interplay with each other to affect the downstream signaling pathways. This grant proposal is to study how cAMP can regulate the dynamics of the exocytotic fusion pore. We will combine molecular biology, cell culture, electrophysiology and live-cell imaging to elucidate the following three questions. Firstly, does cAMP regulate the exocytotic fusion pore directly? Or indirectly by interplaying with Ca2+? Secondly, does cAMP regulate exocytosis via PKA-dependent or PKA-independent pathway? Thirdly, does cAMP modulate the proteins involved in regulated exocytosis, such as the Ca2+ sensor, fusion pore machinery, or the channels mediating cell excitability, to regulate the dynamics of exocytosis? This study will advance our understanding for the mechanisms by which second messengers regulate exocytosis. The results will also provide a new scope to further studies related to neurotransmitter release as well as the relevant neurological diseases.次級信使胞吐作用融合孔Second messagerexocytosisfusion pore