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系統識別號 U0026-0812200911253311
論文名稱(中文) 甲基安非他命引發小鼠場地制約偏好之機制 探討
論文名稱(英文) The mechanism of methamphetamine-induced conditioned place preference in mice
校院名稱 成功大學
系所名稱(中) 細胞生物及解剖學研究所
系所名稱(英) Institute of Cell Biology and Anatomy
學年度 93
學期 1
出版年 94
研究生(中文) 李珮瑜
研究生(英文) Pei-Yu Lee
學號 t9691401
學位類別 碩士
語文別 中文
論文頁數 41頁
口試委員 指導教授-郭余民
召集委員-游一龍
口試委員-楊延光
中文關鍵字 c-Fos  杏仁核  伏隔核  蛋白質合成  非制約刺激  制約刺激  制約場地偏好  甲基安非他命  海馬回 
英文關鍵字 nucleus accumbens  amygdala  hippocampus  c-Fos  protein synthesis  unconditioned stimulus  conditioned stimulus  conditioned place preference  methamphetamine 
學科別分類
中文摘要   之前研究指出學習和記憶在濫用藥物行為中扮演重要的角色。然而藥物引起的記憶之神經受質及其機制至今尚未清楚。制約場地偏好是一檢驗正向刺激作用的實驗模式。制約場地偏好所運用的制約刺激是場地偏好的實驗裝置,而非制約刺激是具酬賞效果的藥物。制約反應則為對能獲得此藥物的場所趨之若鶩。制約場地偏好記憶的形成需要制約刺激和非制約刺激之間的聯結。本研究目的在於探討哪些學習和記憶分子參與了甲基安非他命對C57BL/6J小鼠所建立的制約場地偏好。雖然許多文獻證實長期記憶的形成需要新蛋白質的合成,然而有些報導提出蛋白質合成抑制劑無法影響長期記憶。本研究採用蛋白質合成抑制劑—異黴素,來驗證蛋白質合成在藥物引發的記憶所扮演的角色。結果顯示制約訓練前三十分鐘給與小鼠異黴素(每公斤體重五十毫克,腹腔注射)不會影響甲基安非他命(每公斤體重一毫克,腹腔注射)所建立的制約場地偏好。相對的,事前處理異黴素會減少小鼠對古柯鹼(每公斤體重五毫克,腹腔注射)相關場地的偏好。另外,無論在制約配對二小時之前或之後給與異黴素的處理都無法影響甲基安非他命所建立的制約場地偏好。由於小鼠天性好奇,因此設計了一連串的實驗來驗證甲基安非他命引起場地偏好的原因是否源自於甲基安非他命引起的幻覺剝奪了小鼠對環境線索的認知。實驗結果都指向甲基安非他命引起的場地偏好可能是因為藥物產生的幻覺效應(新鮮感)與制約場地偏好實驗中的對側(非藥物刺激)不平衡所導致,而與非制約刺激較無關連。接著,我利用免疫組織染色偵測c-Fos的表現情形來探究甲基安非他命引起場地偏好的行為中大腦的活化區域。結果顯示杏仁核是甲基安非他命引起場地偏好行為中制約刺激的標的,而伏隔核與非制約刺激有關。總結以上實驗,可得到三個結論:(一) 古柯鹼相關的制約場地偏好需要新合成蛋白質的參與,(二) 甲基安非他命引起的場地偏好可能是藥物產生的幻覺效應所致,而且此現象不需依賴新蛋白質的生成,以及(三) 伏隔核可能與甲基安非他命引起的刺激有關,然而這方面還須進一步的研究才能確立。
英文摘要   Accumulating human studies and animal research evidence showed that learning and memory plays an important role in compulsive drug use behavior. However, neuronal substrates and the mechanism underlying drug-induced memory remain unclear. Conditioned place preference (CPP) is a common laboratory demonstration of positive conditioned incentive effects in which the rewarding drug serves as an unconditioned stimulus (US), while the place-preference apparatus serves as a conditioned stimulus (CS). The conditioned response is place approach behaviors. The formation of CPP memory requires an active memory trace to associate a CS with an US. The objective of this study was to assess the interaction of learning and memory components with methamphetamine-induced place preference in C57BL/6J mice. Although extensive evidence suggested that de novo protein synthesis is required for the formation of long-term memory, some reports described that protein synthesis inhibition did not affect formation of long-term memory. In this study, a protein synthesis inhibitor, anisomycin, was used to examine the role of protein synthesis in forming drug-induced memory. The results showed that pretreated with anisomycin (50 mg/kg, i.p.), 30 min before the conditionings, had no influence on place preference memory established by methamphetamine (1 mg/kg, i.p.). In contrast, cocaine (5 mg/kg, i.p.)-induced place preference was mitigated by anisomycin pretreatment. Neither did anisomycin administration 2 h before nor 2 h after the pairings affect the methamphetamine-induced place preference. Furthermore, since mice tend to explore novelty in nature, a series of experiments were designed to examine the hypothesis: methamphetamine-induced hallucination which distracts their consciousness from attending to the environmental cues may be a reason for elucidating methamphetamine-induced place preference. The findings all indicate that methamphetamine-induced place preference may be primed by the drug-produced hallucinogenic effects. To characterize activated brain regions in methamphetamine-induced place preference, immunohistochemistry was used to detect c-Fos expression. The results suggest that amygdala is a target of CS in methamphetamine-induced place preference. Additionally, nucleus accumbens may be related to US effect. Take together, my studies conclude that (1) cocaine-associated CPP requires the participation of newly synthesized protein, (2) methamphetamine-induced place preference may be primed by the drug-produced hallucinogenic effects and independent of protein synthesis, and (3) nucleus accumbens may be associated with methamphetamine-induced stimuli, however such findings require further investigations.
論文目次 List of Figures ………………………………………iii
Abstract (in Chinese) ………………………………iv
Abstract …………………………………………………vi
1. Introduction …………………………………………1
1.1. Addiction …………………………………………1
1.2. Conditioned place preference …………………2
1.3. Methamphetamine …………………………………3
1.4. Memory and de novo protein synthesis ………4
1.5. Anisomycin …………………………………………5
1.6. c-Fos ………………………………………………6
2. Aims ……………………………………………………8
3. Materials ……………………………………………9
3.1. Animals ……………………………………………9
3.2. Place-preference apparatus ……………………9
3.3. Drugs ………………………………………………9
3.4. Solution preparation …………………………10
3.5. Materials resources ……………………………12
4. Methods ………………………………………………14
4.1. Drug-induced place preference ………………14
4.2. Experimental procedures ………………………14
4.2.1. Effects of anisomycin on memory formation of psychostimulant-induced place preferences …14
4.2.2. Examination on the methamphetamine-produced hallucinogenic effects …………………15
4.2.3. Experiment 7: Expression of c-Fos in methamphetamine-induced place preference ………16
4.3. Tissue preparation for immunohistochemistry ………………………………………………17
4.4. c-Fos immunohistochemistry for free-floating sections ……………………………………18
4.5. Quantitative analysis of c-Fos immunoreactivity ………………………………18
4.6. Statistical analyses ……………………19
5. Results ………………………………………20
5.1. Experiment 1 and 2: Effects of anisomycin on memory formation of psychostimulant-induced place preferences …………………20
5.2. Experiment 3 ~ 6: Examination on the methamphetamine-produced hallucinogenic effects ……………………………………20
5.3. Experiment 7: Expression of c-Fos in methamphetamine-induced place preference ………22
6. Discussion ……………………………………23
References ……………………………………… 28
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