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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 潘建源(Chien-Yuan Pan) | |
dc.contributor.author | Ya-Chu Hsu | en |
dc.contributor.author | 許雅筑 | zh_TW |
dc.date.accessioned | 2021-05-16T16:28:55Z | - |
dc.date.available | 2013-11-18 | |
dc.date.available | 2021-05-16T16:28:55Z | - |
dc.date.copyright | 2013-08-22 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-19 | |
dc.identifier.citation | Ames, J.B., and Lim, S. (2012). Molecular structure and target recognition of neuronal calcium sensor proteins. Biochimica Et Biophysica Acta-General Subjects 1820, 1205-1213.
Aravind, P., Chandra, K., Reddy, P.P., Jeromin, A., Chary, K.V.R., and Sharma, Y. (2008). Regulatory and structural EF-hand motifs of neuronal calcium sensor-1: Mg2+ modulates Ca2+ binding, Ca2+-induced conformational changes, and equilibrium unfolding transitions. Journal of Molecular Biology 376, 1100-1115. Baker, P.F., and Knight, D.E. (1978). Calcium-dependent exocytosis in bovine adrenal-medullary cells with leaky plasma-membranes. Nature 276, 620-622. Barclay, J.W., Morgan, A., and Burgoyne, R.D. (2005). Calcium-dependent regulation of exocytosis. Cell Calcium 38, 343-353. Chandra, K., Ramakrishnan, V., Sharma, Y., and Chary, K.V.R. (2010). Resonance assignments of myristoylated and non-myristoylated neuronal calcium sensor-1(NCS-1) embedded in a membrane. Biomolecular Nmr Assignments 4, 155-158. Colliver, T.L., Hess, E.J., Pothos, E.N., Sulzer, D., and Ewing, A.G. (2000). Quantitative and statistical analysis of the shape of amperometric spikes recorded from two populations of cells. Journal of Neurochemistry 74, 1086-1097. Dason, J.S., Romero-Pozuelo, J., Atwood, H.L., and Ferrus, A. (2012). Multiple Roles for Frequenin/NCS-1 in Synaptic Function and Development. Molecular Neurobiology 45, 388-402. Handley, M.T.W., Lian, L.-Y., Haynes, L.P., and Burgoyne, R.D. (2010). Structural and Functional Deficits in a Neuronal Calcium Sensor-1 Mutant Identified in a Case of Autistic Spectrum Disorder. Plos One 5. LoGiudice, L., and Matthews, G. (2006). The synaptic vesicle cycle: Is kissing overrated? Neuron 51, 676-677. McFerran, B.W., Graham, M.E., and Burgoyne, R.D. (1998). Neuronal Ca2+ sensor 1, the mammalian homologue of frequenin, is expressed in chromaffin and PC12 cells and regulates neurosecretion from dense-core granules. Journal of Biological Chemistry 273, 22768-22772. Pan, C.Y., Jeromin, A., Lundstrom, K., Yoo, S.H., Roder, J., and Fox, A.P. (2002). Alterations in exocytosis induced by neuronal Ca2+ sensor-1 in bovine chromaffin cells. Journal of Neuroscience 22, 2427-2433. Petko, J.A., Kabbani, N., Frey, C., Woll, M., Hickey, K., Craig, M., Canfield, V.A., and Levenson, R. (2009). Proteomic and functional analysis of NCS-1 binding proteins reveals novel signaling pathways required for inner ear development in zebrafish. Bmc Neuroscience 10. Piton, A., Michaud, J.L., Peng, H., Aradhya, S., Gauthier, J., Mottron, L., Champagne, N., Lafreniere, R.G., Hamdan, F.F., Joober, R., et al. (2008). Mutations in the calcium-related gene IL1RAPL1 are associated with autism. Human Molecular Genetics 17, 3965-3974. Weiss, J.L., Archer, D.A., and Burgoyne, R.D. (2000). Neuronal Ca2+ sensor-1/frequenin autocrine pathway regulating Ca2+ bovine adrenal chromaffin cells. Journal of Biological Chemistry 275, 40082-40087. Weiss, J.L., Hui, H., and Burgoyne, R.D. (2010). Neuronal Calcium Sensor-1 Regulation of Calcium Channels, Secretion, and Neuronal Outgrowth. Cellular and Molecular Neurobiology 30, 1283-1292. Burgoyne, R.D. (2007). Neuronal calcium sensor proteins: generating diversity in neuronal Ca2+ signalling. Nature Reviews Neuroscience 8, 182-193. Jahn, R., and Fasshauer, D. (2012). Molecular machines governing exocytosis of synaptic vesicles. Nature 490, 201-207. Wang, Ying Shiuan (2010). Effects of NCS-1 and Auxillin-1 on Exo / endocytosis. Master Thesis. Institute of Zoology, National Taiwan University , Taipei Chen, Yi Fan (2012). Effects of NCS-1 and Auxillin-1 on Neurotransmission. Master Thesis. Institute of Zoology, National Taiwan University , Taipei | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/6419 | - |
dc.description.abstract | 鈣離子結合蛋白的家族種類眾多,其中包含具有EF hands結構,可與鈣離子結合的鈣離子感應蛋白,如neuronal calcium semsor-1 (NCS-1),它的結構含有N端myristoylation及三個有功能的EF hands。前人的研究指出,NCS-1會影響分泌膜囊的回收再利用,但詳細機制尚未釐清。我們在牛腎上腺髓質細胞中表現NCS-1,運用安培法測量分泌出的神經傳遞物質以及影像技術來偵測胞內鈣離子變化,以研究高鉀刺激所造成突觸小泡釋放的機轉。我們發現,相較於對照組,表現NCS-1 的細胞,其胞吐作用沒有顯著變化;表現NCS-1G2A (無法結合細胞膜)或NCS-1R102Q (發現在某些自閉症患者)的細胞,其胞吐程度降低,氧化電流和電量均變小,顯示有抑制胞吐作用的現象及突觸小泡內的傳導物質可能減少;而NCS-1E120Q (無法結合鈣離子) 則增加胞吐電量。胞内鈣離子濃度變化的結果顯示,過度表現NCS-1G2A的細胞有顯著較低的鈣離子反應,而表現NCS-1與其他變異的實驗組,則與控制組細胞相近。這些結果顯示,調節胞吐作用的功能與結合到膜上能力有很大相關;而失去鈣離子結合能力,並不影響NCS-1調節的作用。因此NCS-1可能參與在調控分泌膜囊的回收再利用機轉中,並在神經突觸的長期變化,扮演重要的角色。 | zh_TW |
dc.description.abstract | Many proteins have a Ca2+ binding capability and involved in various physiological activities. Calcium sensor protein family belongs to the calcium binding protein group and has EF hands for Ca2+ binding. Neuronal calcium sensor-1 (NCS-1) has an N-terminal myristoylation site, one cryptic EF hand and 3 functional EF hands. NCS-1 overexpression affects the vesicle recycling, however, the mechanism is not clear. In this report, we monitor the catecholamine release and [Ca2+]i evoked by high K+ depolarization to characterize the roles of NCS-1 in the stimulus-secretion coupling in bovine chromaffin cells. We found that cells overexpressing NCS-1 had similar exocytosis level as the control group expressing GFP; cells overexpressing NCS-1G2A (no plasma membrane anchoring capability) or NCS-1R102Q (identified in autism patients) had less exocytosis events, and smaller oxidation currents; in contrast, cells overexpressing NCS-1E120Q (loses Ca2+-binding ability) increased the exocytosis level. For the intracellular Ca2+ response, all constructs caused similar responses as the control group except NCS-1G2A which had a smaller elevation than the control group. These results indicated that the plasma membrane targeting capability is important for the regulatory role of the NCS-1 in the stimulus-secretion coupling; however, losing the Ca2+-binding ability does not affect the regulatory role of the NCS-1. In summary, NCS-1 may be involved in regulating the synaptic vesicle recycling and playing an important role in long term synapse plasticity changes. | en |
dc.description.provenance | Made available in DSpace on 2021-05-16T16:28:55Z (GMT). No. of bitstreams: 1 ntu-102-R00b41013-1.pdf: 3601817 bytes, checksum: c0f095171e608a9f3053e8de59cb4738 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 誌謝 i
摘要 ii Abstract iii 1.Introduction 1 1.1 Synaptic Transmission 1 1.2 Vesicle Cycle 3 1.3 Calcium 4 1.4 Neuronal Calcium Sensor-1 (NCS-1) 5 1.5 Approaches 6 1.6 Aims 8 2. Materials and Methods 9 2.1 Solutions 9 2.2 Primary bovine chromaffin cell culture 10 2.3 Plasmid of NCS-1 and its mutations 11 2.4 LTX transfection 12 2.5 Stimulation and Amperometry 13 2.6 Calcium imaging technique 14 2.7 Data Analysis 14 3. Results 16 3.1 The traces of amperometry 16 3.2 Cells overexpressing NCS-1 maintains the exocytosis; cells overexpressing NCS-1G2A or NCS-1R102Q down-regulate the exocytosis; cells overexpressing NCS-1E120Q have an elevated exocytosis level. 17 3.3 Cells overexpressing NCS-1E120Q have long vesicle fusion time. 18 3.4 Cells overexpressing NCS-1G2A or NCS-1R102Q have relative infrequent exocytosis in the 3rd sweep. 20 3.5 Cells overexpressing NCS-1G2A have a relative uniform distribution on the exocytosis events 21 3.6 The traces of calcium imaging 21 3.7 Cells overexpressing NCS-1G2A reduces calcium response 22 3.8 The relationship between Ca2+ response and exocytosis 23 4. Discussion 24 4.1 Cells overexpressing NCS-1 maintains the exocytosis activities 24 4.2 Cells overexpressing NCS-1G2A or NCS-1R102Q down-regulate the exocytosis 25 4.3 Cells overexpressing NCS-1E120Q have an elevated exocytosis level 26 4.4 The timing of exocytosis events 27 4.5 NCS-1G2A overexpression reduces Ca2+ response 27 4.6 NCS-1 keeps the machinery between exocytosis and Ca2+ response. 28 4.7 Internal Ca2+ store may mediate the function of NCS-1 29 4.8 Suggestions for improvements 30 5. Conclusion 31 6. References 32 Schemes………………………………………………………………………………35 Figures………………………………………………………………………………..39 | |
dc.language.iso | en | |
dc.title | 牛腎上腺嗜鉻細胞中NCS-1維持了正常的胞吐活動 | zh_TW |
dc.title | NCS-1 Maintains the Normal Exocytotic Activities in Bovine Chromaffin Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 戴晶瑩(Chin-Yin Tai),王致恬(Chih-Tien Wang),陳示國(ShihKuo Chen) | |
dc.subject.keyword | NCS-1,腎上腺髓質細胞,胞吐作用,胞吞作用,分泌膜囊的回收再利用, | zh_TW |
dc.subject.keyword | NCS-1,chromaffin cell,exocytosis,endocytosis,synaptic vesicle recycling, | en |
dc.relation.page | 48 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2013-08-19 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 動物學研究所 | zh_TW |
顯示於系所單位: | 動物學研究所 |
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