Please use this identifier to cite or link to this item:
https://ah.lib.nccu.edu.tw/handle/140.119/66497| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 趙知章 | zh_TW |
| dc.contributor.author | 洪禎廷 | zh_TW |
| dc.creator | 洪禎廷 | zh_TW |
| dc.date | 2013 | en_US |
| dc.date.accessioned | 2014-06-04T06:45:13Z | - |
| dc.date.available | 2014-06-04T06:45:13Z | - |
| dc.date.issued | 2014-06-04T06:45:13Z | - |
| dc.identifier | G0100754007 | en_US |
| dc.identifier.uri | http://nccur.lib.nccu.edu.tw/handle/140.119/66497 | - |
| dc.description | 碩士 | zh_TW |
| dc.description | 國立政治大學 | zh_TW |
| dc.description | 神經科學研究所 | zh_TW |
| dc.description | 100754007 | zh_TW |
| dc.description | 102 | zh_TW |
| dc.description.abstract | 蛋白激酶CK2（Casine kinase 2）為四單體所構成，針對配受質蛋白之絲胺酸或蘇胺酸位置進行磷酸化，先前研究已經發現在紋狀體腦區之CK2的表現量與活性皆高於大腦中其餘腦區，而紋狀體腦區主要神經細胞為-氨基丁酸神經元（GABAergic neurons）的medium spiny neuron（MSN），會受到來自黑質多巴胺神經細胞（dopaminergic neurons）的調控。此外，DARPP-32（dopamine- and cAMP-regulated phosphoprotein, Mr 32 kDA）蛋白亦被發現大量表現於在MSN細胞中，且為CK2之受質蛋白質。雖然CK2已被證實參與多巴胺神經元的神經保護機制，但是否參與MSN細胞對運動行為調控之生理機制仍未清楚。由於已有研究發現施予1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）藥物處理造成黑質-紋狀體腦區受損之老鼠腦內-氨基丁酸（GABA）的生合成酵素─麩胺酸脫羧酵素67（GAD67）表現量與正常老鼠不同，因此本論文研究的主題擬在大鼠實驗模式中利用MPP+造成投射至紋狀體之多巴胺神經細胞受損，探討當多巴胺調控紋狀體神經細胞能力缺失的狀態下，MSN細胞之CK2、DARPP-32和GAD蛋白表現與動物運動行為之相關性。\r\n實驗結果發現，直接於紋狀體給予1-甲基-4-苯基碘化啶 （MPP+ Iodide）皆會造成CK2、DARPP-32以及GAD67之蛋白質含量的減少，多巴胺及其代謝物和GABA等神經化學傳遞物質亦有減少的現象；另外，在MPP+給予前分別操弄CK2或DARPP-32 胺基酸Ser102磷酸化的表現，皆會改變GAD67蛋白質含量與黑質酪胺酸羥化酶（Tyrosine Hydroxylase, TH）蛋白質含量，同時神經化學傳遞物質的含量或代謝亦有改變。由現有之結果推測CK2/DARPP-32/GAD67細胞訊息傳遞機制可能參與巴金森氏症運動行為失常之細胞層面的調控。 | zh_TW |
| dc.description.abstract | Protein kinase CK2 is a heterotetrameric and serine/threonine protein kinase. Its protein levels and activity are found to be elevated in the striatum when compared to other brain areas. CK2 is known to involve in the neuroprotective effects of dopaminergic neurons, whether it also regulates the neuronal function relative to motor behaviors is still unclear. DARPP-32 protein is known as one of the substrates for CK2 and is highly expressed in the GABAergic medium spiny neurons (MSN) responsible for dopamine stimulation in the striatum. Furthermore, other studies have indicated that the expression of glutamic acid decarboxylase 67 (GAD67) mRNA and protein was different in the striatum of MPTP vs. naïve animals, which is one of the enzymes responsible for the synthesis of neurotransmitter GABA. In the present study, we observed that the parallel changes in protein levels of CK2, DARPP-32 and GAD67 in the striatum and TH in the substantia nigra of MPP+-treated. We also found that manipulation of CK2 or DARPP-32 gene expression aggravated the MPP+-induced neuropathological dificts. The present results suggest that CK2/DARPP-32/GAD67 signaling pathway might involve in the cellular mechanism of motor-deficit in Parkinson’s disease. | en_US |
| dc.description.tableofcontents | 謝誌 I\r\n中文摘要 II\r\nAbstract III\r\n目錄 IV\r\n表次 VI\r\n圖次 VII\r\n縮寫對照表 VIII\r\n第一章 緒論 1\r\n第一節 巴金森氏症（Parkinson’s Disease）病理機制 2\r\n第二節 左旋多巴胺誘發之運動障礙（L-DOPA-induced deskynesia） 5\r\n第三節 蛋白激酶CK2（Protein kinase CK2, Casein Kinase 2） 8\r\n第四節 DARPP-32（Dopamine- and cAMP-regulated phosphoprotein, Mr 32 kDa） 10\r\n第五節 麩胺酸脫羧酵素67（Glutamic Acid Decarboxylase 67, GAD67） 13\r\n第六節 論文研究目的與策略 14\r\n第二章 實驗材料與方法 15\r\n第一節 實驗動物 16\r\n第二節 立體定位手術（stereotaxic surgery） 16\r\n第三節 質體DNA之萃取 16\r\n第四節 CK2a siRNA質體DNA與的轉染作用（transfection）以及MPP+毒性作用 17\r\n一、小干擾RNA轉染作用 17\r\n二、質體DNA與聚乙烯亞胺（polyethylenimine, PEI）之製備與轉染反應 17\r\n三、紋狀體之轉染作用及注射質體DNA/PEI混合液 18\r\n四、1-甲基-4-苯基吡啶（MPP+）之毒性作用 19\r\n第五節 藥物微量注射程序 19\r\n一、MPP+微量注射 19\r\n二、CK2a siRNA與MPP+微量注射 19\r\n三、DARPP-32 S102A或S102D與MPP+微量注射 19\r\n第六節 西方墨點法（western blot） 20\r\n一、紋狀體及黑質體組織擷取 20\r\n二、蛋白質萃取 20\r\n三、蛋白質濃度測定及標準曲線製作 20\r\n四、蛋白質樣本配置 21\r\n五、鑄膠及聚丙烯醯胺凝膠電泳（sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE） 21\r\n六、轉漬（transfer） 21\r\n七、免疫轉印（immunoblotting） 21\r\n第七節 高效能液相層析法（High Performance Liquid Chromatography, HPLC） 22\r\n一、神經傳導物質之萃取 22\r\n二、流動相（mobile phase）配置 23\r\n三、高效能液相層析法 23\r\n第八節 統計分析 23\r\n第三章 實驗結果 24\r\n第一節 MPP+對紋狀體蛋白質含量和神經化學傳遞物質含量的影響 25\r\n第二節 轉染CK2a siRNA與MPP+與對紋狀體蛋白質含量和神經化學傳遞物質含量的影響 32\r\n第三節 轉染CK2 a siRNA與MPP+對黑質蛋白質含量和神經化學傳遞物質含量以及對行為的影響 37\r\n第四節 轉染突變型DARPP-32 S102A或S102D質體DNA與MPP+對紋狀體蛋白質含量和神經化學傳遞物質含量的影響 44\r\n第五節 轉染突變型DARPP-32 S102A或S102D質體DNA與MPP+對黑質蛋白質和神經化學傳遞物質含量以及對行為的影響 49\r\n第四章 討 論 60\r\n結 論 65\r\n參考文獻 66 | zh_TW |
| dc.language.iso | en_US | - |
| dc.source.uri | http://thesis.lib.nccu.edu.tw/record/#G0100754007 | en_US |
| dc.subject | 紋狀體 | zh_TW |
| dc.subject | MSN細胞 | zh_TW |
| dc.subject | 1-甲基-4-苯基-1,2,3,6-四氢吡啶碘化物 | zh_TW |
| dc.subject | 蛋白激酶CK2 | zh_TW |
| dc.subject | DARPP-32蛋白 | zh_TW |
| dc.subject | 麩胺酸脫羧酵素67 | zh_TW |
| dc.subject | gamma-丁氨基酪酸 | zh_TW |
| dc.subject | 多巴胺 | zh_TW |
| dc.subject | striatum | en_US |
| dc.subject | medium spiny neuron | en_US |
| dc.subject | protein kinase CK2 | en_US |
| dc.subject | DARPP-32 | en_US |
| dc.subject | glutamic acid decarboxylase 67 | en_US |
| dc.subject | gamma-aminobutyric acid | en_US |
| dc.subject | dopamine | en_US |
| dc.title | 1-甲基-4-苯基碘化啶對大鼠紋狀體神經細胞中CK2/DARPP-32/GAD67訊息傳遞表現及 神經生理功能之影響 | zh_TW |
| dc.title | Effect of MPP+ on CK2/DARPP-32/GAD67 signaling pathway and neurophysiological function in the striatum of rats | en_US |
| dc.type | thesis | en |
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