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請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32264
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dc.contributor.advisor潘建源(Chien-Yuan Pan)
dc.contributor.authorRuo-Lin Kangen
dc.contributor.author康若琳zh_TW
dc.date.accessioned2021-06-13T03:39:38Z-
dc.date.available2006-07-29
dc.date.copyright2006-07-29
dc.date.issued2006
dc.date.submitted2006-07-25
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32264-
dc.description.abstract鈣離子結合蛋白 (Ca2+-binding protein 1-5, CaBP1-5) 是一群與calmodulin有高度相似性結構的蛋白質,目前僅知CaBP1及CaBP4會調控鈣離子通道的活性,而其它蛋白的功能則尚不清楚。本篇報告中,我們以PCR技術從大鼠 (Rattus norvegicus) 胚胎的大腦組織中選殖出CaBP5基因,再予以表現於兩類型興奮性細胞中,來探討其對刺激-分泌耦合機轉的調控。細胞螢光染色結果顯示,CaBP5-EYFP主要分布於細胞質中,有些則會集中於細胞核。利用細胞電生理技術,我們發現 CaBP5的大量表現會使神經細胞的鈣離子通道的開啟降低至較負電位,失去第一個 EF-hand正常功能的突變型 (5EF1) 則沒有作用。但在牛腎上腺嗜鉻細胞 (bovine chromaffin cell) 中,CaBP5對其鈣離子通道沒有同樣影響,藉由測量細胞膜電容的變化發現CaBP5的表現,也不會影響胞吐和胞吞作用。在牛腎上腺嗜鉻細胞中,CaBP5對鈣離子通道沒有同樣的作用,也沒有對胞吐 (exocytosis) 和胞吞作用 (endocytosis) 造成明顯的影響;另外,CaBP5對鈉離子電流沒有影響,但5EF1的表現增強了鈉離子電流。以上觀察顯示CaBP5可能參與神經細胞膜的鈣離子電流調控,而對鈉離子電流的影響不明顯;但在牛腎上腺嗜鉻細胞,5EF1增強了鈉離子電流的大小。zh_TW
dc.description.abstractCa2+ -binding proteins 1-5 (CaBP1-5) are a group of proteins with structure similar to calmodulin. CaBP1 and CaBP4 have been reported to modulate the Ca2+ currents and synaptic plasticity of neurons; however, functions of other proteins have not been extensively investigated. In this report, functions of CaBP5 in excitable cells were studied. CaBP5 was cloned from rat (Rattus norvegicus) embryonic brain tissue and inserted into pEYFP-N1 for expression. Normally it was localized in the cytosol, but it may concentrat in the nucleus in some cells. We applied whole-cell patch clamping technique to monitor the electrophysiological properties of excitable cells. The results show that CaBP5 shifted the activation of Ca2+ channels to hyperpolarized voltages in primary neuron culture, but mutant CaBP5 with the first nonfunctional EF-hand didn’t have the same effect. When CaBP5 was expressed in cultured bovine adrenal chromaffin cells, neither the Ca2+ currents nor the exocytosis was affected, but the Na+ currents were enhanced. These observations indicate that CaBP5 may be involved in regulation of Ca2+ channels in neurons but enhanced Na+ currents in chromaffin cells.en
dc.description.provenanceMade available in DSpace on 2021-06-13T03:39:38Z (GMT). No. of bitstreams: 1
ntu-95-R92b41017-1.pdf: 461945 bytes, checksum: 6aeab7c4211b93b91d36dcae610cbc0c (MD5)
Previous issue date: 2006
en
dc.description.tableofcontents1.摘要
1.1 中文摘要..................................................... 1
1.2 英文摘要.....................................................2
2.前言
2.1 Calmodulin...................................................3
2.2 鈣離子結合蛋白...............................................4
2.3 實驗目標.....................................................5
3. 材料與方法
3.1 溶液........................................................ 7
3.2 RNA 製備與 cDNA 合成..................................... 7
3.3 CaBP5 轉接入 pEYFP-N1 載體與核苷酸點突變合成.............8
3.4 神經細胞與牛腎上腺嗜鉻細胞培養...............................9
3.5 基因轉殖 (Gene Transfection) 與細胞螢光染色................... 9
3.6 細胞電生理技術..............................................10
3.7 統計分析....................................................11
4. 結果
4.1 CaBP5 表現於大鼠的視網膜與胎鼠的大腦組織中................ 12
4.2 CaBP5 主要是均勻分布於神經細胞中,少數集中於細胞核內...... 12
4.3 CaBP5使神經細胞的鈣離子通道的開啟降低至較負電位...........13
4.4 CaBP5對神經細胞的鈉離子電流沒有影響.......................13
4.5 CaBP5 對牛腎上腺嗜鉻細胞的鈣離子電流沒有同樣影響.......... 14
4.6 CaBP5對牛腎上腺嗜鉻細胞的鈉離子電流有增強的效果........... 14
4.7 CaBP5 對牛腎上腺嗜鉻細胞的胞吐和胞吞作用沒有影響.......... 15
5. 討論...........................................................16
6. 致謝...........................................................20
7. 圖表...........................................................21
8. 參考文獻...................................................... 32
dc.language.isozh-TW
dc.subject牛腎上腺嗜鉻細胞zh_TW
dc.subject分泌作用zh_TW
dc.subject鈉離子通道zh_TW
dc.subject鈣離子通道zh_TW
dc.subject鈣離子結合蛋白zh_TW
dc.subject神經細胞zh_TW
dc.subjectvoltage-gated Na+ chabnnelen
dc.subjectendocytosisen
dc.subjectCa2+ -binding protein 5en
dc.subjectneuronen
dc.subjectbovine chromaffin cellen
dc.subjectvoltage-gated Ca2+ channelen
dc.subjectexocytosisen
dc.title鈣離子結合蛋白5在興奮性細胞的生理功能研究zh_TW
dc.titleFunctions of Ca2+-Binding Protein 5 in Excitable Cellsen
dc.typeThesis
dc.date.schoolyear94-2
dc.description.degree碩士
dc.contributor.oralexamcommittee陳建璋(Chien-Chang Chen),閔明源(Ming-Yuan Min),林崇智(Chung-Chih Lin)
dc.subject.keyword鈣離子結合蛋白,神經細胞,牛腎上腺嗜鉻細胞,鈣離子通道,鈉離子通道,分泌作用,zh_TW
dc.subject.keywordCa2+ -binding protein 5,neuron,bovine chromaffin cell,voltage-gated Ca2+ channel,voltage-gated Na+ chabnnel,exocytosis,endocytosis,en
dc.relation.page35
dc.rights.note有償授權
dc.date.accepted2006-07-27
dc.contributor.author-college生命科學院zh_TW
dc.contributor.author-dept動物學研究研究所zh_TW
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