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系統識別號 U0026-2901201612204800
論文名稱(中文) 探討杏仁核中甲基安非他命相關記憶去穩定化過程的機制
論文名稱(英文) To Study the Mechanism of Destabilization of Methamphetamine-related Memory in Amygdala
校院名稱 成功大學
系所名稱(中) 基礎醫學研究所
系所名稱(英) Institute of Basic Medical Sciences
學年度 104
學期 1
出版年 104
研究生(中文) 余泱蓉
研究生(英文) Yang-Jung Yu
學號 S58981206
學位類別 博士
語文別 英文
論文頁數 83頁
口試委員 指導教授-簡伯武
召集委員-吳豐森
口試委員-黃阿敏
口試委員-符文美
口試委員-曾清俊
口試委員-黃金鼎
中文關鍵字 甲基安非他命  再固化  杏仁核  鈣調神經磷酸酶  去穩定化 
英文關鍵字 Methamphetamine  Reconsolidation  Amygdala  Calcineurin  Destabilization 
學科別分類
中文摘要 重複的藥物攝取導致神經方面持久性的改變,其所導致的強迫性藥物使用行為則稱為成癮。藥物戒斷過程中容易產生復發,主要因素是藥物記憶的存在。記憶再固化過程能讓固化記憶更新訊息並更穩固存在,其中包含提取引發的去穩定化及再穩定化兩個步驟。提取引發的去穩定化能使固化記憶變成不穩定狀態而容易被破壞,而必須經過再穩定化,才能使不穩定記憶再次穩固並持久存在。由於提取後記憶的不穩定,因此,近來許多研究著重在破壞藥物記憶的再固化過程,以達到破壞藥物記憶,減少藥物渴求的表現。然而,目前對於如何破壞藥物記憶再固化的機制尚不明確,以及提取後記憶轉為不穩定狀態的機制也尚未知。過去研究也發現杏仁核參與環境線索與藥物連結性記憶的形成。然而,杏仁核在調控環境線索與藥物記憶形成所扮演的參與機制仍需要更多討論。在這裡,我們使用場地制約偏好模式探討杏仁核中藥物相關記憶去穩定化的細胞機制。我們施予小鼠甲基安非他命三天制約訓練,使其產生回饋反應及藥物記憶。在場地偏好測試(記憶提取)後1小時給予茴香黴素,發現能顯著破壞藥物記憶的表現。另外,茴香黴素對甲基安非他命場地制約小鼠在側邊杏仁核所造成的突觸膜上麩氨酸受體增加、麩氨酸受體次單位GluR1 serine845磷酸化(p-GluR1-Ser845)增加、突觸數目增加及興奮性突觸傳遞的增加有抑制的效果。然而,在雙邊側杏仁核微量注入阻斷麩氨酸受體內吞作用的合成胜肽(Tat-GluR23Y)、N-甲基-D-天冬氨酸受體阻抗劑(MK801)、NR2B次單的N-甲基-D-天冬氨酸受體阻抗劑(ifenprodil)、鈣調神經磷酸酶抑制劑(FK50及cyclosporin A)及蛋白質脫磷酸化酵素1抑制劑(okadaic acid及calyculin A),能阻止茴香黴素對甲基安非他命記憶小鼠在行為及分子層面上的破壞效果。由這些結果推測記憶提取能導致NR2B次單的N-甲基-D-天冬氨酸受體活化,其能使鈣離子流入細胞內。細胞內鈣離子的增加能刺激鈣調神經磷酸酶,並活化蛋白質脫磷酸化酵素1,然後p-GluR1-ser845去磷酸化,進而發生麩氨酸受體內吞作用及藥物記憶的去穩定化。
英文摘要 Repetitive drug taking induces neural long-lasting changes and results in compulsive drug-seeking behavior known as addiction. Drug withdrawal is easy to relapse which may result from drug memory. Thus, recent literatures focus on disrupting these memories to reduce drug seeking. Reconsolidation involves retrieval-induced destabilization and restabilization processes in order for memories to be updated. Retrieval-induced destabilization render memories susceptible to disruption by amnestic agents. Previous studies revealed that the amygdala is involved in the drug memory. However, the role of the amygdala in the regulation of drug memory reconsolidation and its participation in the mechanisms still need more discussion. Here, we used conditioned place preference (CPP) procedure in mice to delineate the cellular mechanism underlying the destabilization of drug-related memory in amygdala. Methamphetamine conditioning (MeAM) for 3 days in mice (2 mg/kg, i.p.) induced a significant reward effect. Mice then received anisomycin (ANI) within 1 hour after retrieval to disrupt reconsolidation of MeAM-related memory. ANI reversed MeAM-induced increase in the level of surface AMPA receptors (AMPARs), phosphorylation of GluR1-Serine845 (p-GluR1-Ser845), dendritic spines and the excitatory synaptic transmission in the basal lateral amygdala (BLA). The effects of ANI were prevented by bilateral injection of synthetic peptide that blocked AMPAR endocytosis, N-methyl-D-aspartate receptor (NMDAR) antagonist, NR2B-containing NMDAR antagonist, calcineurin inhibitors and protein phosphatase 1 inhibitors. These results suggest that memory retrieval induces activation of NR2B-containing NMDARs which allows the influx of Ca2+. Increased intracellular Ca2+ stimulates calcineurin leading to dephosphorylation and inactivation of inhibitor 1, and activation of protein phosphatase 1. Protein phosphatase 1 then dephosphorylates p-GluR1-ser845 to elicit AMPAR endocytosis and destabilization of drug-related memory.
論文目次 Contents
Abstract in Chinese...................................I
Abstract in English...................................IV
誌謝..................................................VII
Contents..............................................IX
List of Figures......................................XI
Abbreviations........................................XIV
Introduction.........................................1
Material and Methods.................................8
Results..............................................17
Establishment of methamphetamine-induced conditioned place preference.....................................18
Methamphetamine-paired mice exhibit increased surface AMPA receptors in amygdala...........................19
Anisomycin disrupts MeAM-related memory, which required retrieval............................................20
AMPA receptor endocytosis in the amygdala underlies anisomycin-mediated disruption of MeAM-related memory................................................23
Anisomycin-mediated disruption of MeAM-related memory is blocked by NR2B-NMDA receptor antagonist..............27
Calcineurin is involved in destabilization of MeAM-related memory........................................30
Discussion............................................33
Conclusion and Future perspectives....................41
References............................................44
Figures and Legends...................................59
Curriculum Vitae and Publications.....................83
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