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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳小梨 | |
dc.contributor.author | Ssu-Yu Lin | en |
dc.contributor.author | 林思瑜 | zh_TW |
dc.date.accessioned | 2021-05-17T09:15:21Z | - |
dc.date.available | 2015-09-19 | |
dc.date.available | 2021-05-17T09:15:21Z | - |
dc.date.copyright | 2012-09-19 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-08-10 | |
dc.identifier.citation | Arai M, Alpert NR, MacLennan DH, Barton P, Periasamy M (1993) Alterations in sarcoplasmic reticulum gene expression in human heart failure. A possible mechanism for alterations in systolic and diastolic properties of the failing myocardium. Circulation research 72: 463-469
Arber S, Hunter JJ, Ross J, Jr., Hongo M, Sansig G, Borg J, Perriard JC, Chien KR, Caroni P (1997) MLP-deficient mice exhibit a disruption of cardiac cytoarchitectural organization, dilated cardiomyopathy, and heart failure. Cell 88: 393-403 Bahler M, Rhoads A (2002) Calmodulin signaling via the IQ motif. FEBS letters 513: 107-113 Barry WH, Bridge JH (1993) Intracellular calcium homeostasis in cardiac myocytes. Circulation 87: 1806-1815 Boateng SY, Belin RJ, Geenen DL, Margulies KB, Martin JL, Hoshijima M, de Tombe PP, Russell B (2007) Cardiac dysfunction and heart failure are associated with abnormalities in the subcellular distribution and amounts of oligomeric muscle LIM protein. American journal of physiology Heart and circulatory physiology 292: H259-269 Broderick MJ, Winder SJ (2005) Spectrin, alpha-actinin, and dystrophin. Advances in protein chemistry 70: 203-246 Bruce RA (1974) Methods of exercise testing. Step test, bicycle, treadmill, isometrics. The American journal of cardiology 33: 715-720 Campbell AM, Kessler PD, Sagara Y, Inesi G, Fambrough DM (1991) Nucleotide sequences of avian cardiac and brain SR/ER Ca(2+)-ATPases and functional comparisons with fast twitch Ca(2+)-ATPase. Calcium affinities and inhibitor effects. The Journal of biological chemistry 266: 16050-16055 Chang SW, Tsao YP, Lin CY, Chen SL (2011) NRIP, a novel calmodulin binding protein, activates calcineurin to dephosphorylate human papillomavirus E2 protein. Journal of virology 85: 6750-6763 Chen PH, Tsao YP, Wang CC, Chen SL (2008) Nuclear receptor interaction protein, a coactivator of androgen receptors (AR), is regulated by AR and Sp1 to feed forward and activate its own gene expression through AR protein stability. Nucleic acids research 36: 51-66 Chien KR (1999) Stress pathways and heart failure. Cell 98: 555-558 Chiu C, Bagnall RD, Ingles J, Yeates L, Kennerson M, Donald JA, Jormakka M, Lind JM, Semsarian C (2010) Mutations in alpha-actinin-2 cause hypertrophic cardiomyopathy: a genome-wide analysis. Journal of the American College of Cardiology 55: 1127-1135 Cooper JA, Schafer DA (2000) Control of actin assembly and disassembly at filament ends. Current opinion in cell biology 12: 97-103 Cukovic D, Lu GW, Wible B, Steele DF, Fedida D (2001) A discrete amino terminal domain of Kv1.5 and Kv1.4 potassium channels interacts with the spectrin repeats of alpha-actinin-2. FEBS letters 498: 87-92 Frank D, Kuhn C, Katus HA, Frey N (2006) The sarcomeric Z-disc: a nodal point in signalling and disease. J Mol Med (Berl) 84: 446-468 Frey N, Olson EN (2003) Cardiac hypertrophy: the good, the bad, and the ugly. Annual review of physiology 65: 45-79 Gokhin DS, Fowler VM (2011) Tropomodulin capping of actin filaments in striated muscle development and physiology. Journal of biomedicine & biotechnology 2011: 103069 Hart MC, Cooper JA (1999) Vertebrate isoforms of actin capping protein beta have distinct functions In vivo. The Journal of cell biology 147: 1287-1298 Hasenfuss G (1998) Alterations of calcium-regulatory proteins in heart failure. Cardiovascular research 37: 279-289 Hassel D, Dahme T, Erdmann J, Meder B, Huge A, Stoll M, Just S, Hess A, Ehlermann P, Weichenhan D, Grimmler M, Liptau H, Hetzer R, Regitz-Zagrosek V, Fischer C, Nurnberg P, Schunkert H, Katus HA, Rottbauer W (2009) Nexilin mutations destabilize cardiac Z-disks and lead to dilated cardiomyopathy. Nature medicine 15: 1281-1288 Heineke J, Molkentin JD (2006) Regulation of cardiac hypertrophy by intracellular signalling pathways. Nature reviews Molecular cell biology 7: 589-600 Hemler ME (1999) Dystroglycan versatility. Cell 97: 543-546 Hunter JJ, Chien KR (1999) Signaling pathways for cardiac hypertrophy and failure. The New England journal of medicine 341: 1276-1283 Kim Y, Phan D, van Rooij E, Wang DZ, McAnally J, Qi X, Richardson JA, Hill JA, Bassel-Duby R, Olson EN (2008) The MEF2D transcription factor mediates stress-dependent cardiac remodeling in mice. The Journal of clinical investigation 118: 124-132 Knoll R, Buyandelger B, Lab M (2011) The sarcomeric Z-disc and Z-discopathies. Journal of biomedicine & biotechnology 2011: 569628 Littlefield R, Almenar-Queralt A, Fowler VM (2001) Actin dynamics at pointed ends regulates thin filament length in striated muscle. Nature cell biology 3: 544-551 Littlefield R, Fowler VM (1998) Defining actin filament length in striated muscle: rulers and caps or dynamic stability? Annual review of cell and developmental biology 14: 487-525 Marks AR (2003) Calcium and the heart: a question of life and death. The Journal of clinical investigation 111: 597-600 Mohapatra B, Jimenez S, Lin JH, Bowles KR, Coveler KJ, Marx JG, Chrisco MA, Murphy RT, Lurie PR, Schwartz RJ, Elliott PM, Vatta M, McKenna W, Towbin JA, Bowles NE (2003) Mutations in the muscle LIM protein and alpha-actinin-2 genes in dilated cardiomyopathy and endocardial fibroelastosis. Molecular genetics and metabolism 80: 207-215 Ohrtman J, Ritter B, Polster A, Beam KG, Papadopoulos S (2008) Sequence differences in the IQ motifs of CaV1.1 and CaV1.2 strongly impact calmodulin binding and calcium-dependent inactivation. The Journal of biological chemistry 283: 29301-29311 Ono S (2010) Dynamic regulation of sarcomeric actin filaments in striated muscle. Cytoskeleton (Hoboken) 67: 677-692 Pieske B, Maier LS, Bers DM, Hasenfuss G (1999) Ca2+ handling and sarcoplasmic reticulum Ca2+ content in isolated failing and nonfailing human myocardium. Circulation research 85: 38-46 Reed PW, Lardy HA (1972) A23187: a divalent cation ionophore. The Journal of biological chemistry 247: 6970-6977 Richardson P, McKenna W, Bristow M, Maisch B, Mautner B, O'Connell J, Olsen E, Thiene G, Goodwin J, Gyarfas I, Martin I, Nordet P (1996) Report of the 1995 World Health Organization/International Society and Federation of Cardiology Task Force on the Definition and Classification of cardiomyopathies. Circulation 93: 841-842 Sadeghi A, Doyle AD, Johnson BD (2002) Regulation of the cardiac L-type Ca2+ channel by the actin-binding proteins alpha-actinin and dystrophin. American journal of physiology Cell physiology 282: C1502-1511 Sandow A (1952) Excitation-contraction coupling in muscular response. The Yale journal of biology and medicine 25: 176-201 Saucerman JJ, Bers DM (2012) Calmodulin binding proteins provide domains of local Ca2+ signaling in cardiac myocytes. Journal of molecular and cellular cardiology 52: 312-316 Sjoblom B, Salmazo A, Djinovic-Carugo K (2008) Alpha-actinin structure and regulation. Cellular and molecular life sciences : CMLS 65: 2688-2701 Smith GL, Steele DS (1998) Measurement of SR Ca2+ content in the presence of caffeine in permeabilised rat cardiac trabeculae. Pflugers Archiv : European journal of physiology 437: 139-148 Taylor MP, Koyuncu OO, Enquist LW (2011) Subversion of the actin cytoskeleton during viral infection. Nature reviews Microbiology 9: 427-439 Tsai TC, Lee YL, Hsiao WC, Tsao YP, Chen SL (2005) NRIP, a novel nuclear receptor interaction protein, enhances the transcriptional activity of nuclear receptors. The Journal of biological chemistry 280: 20000-20009 Vikstrom KL, Bohlmeyer T, Factor SM, Leinwand LA (1998) Hypertrophy, pathology, and molecular markers of cardiac pathogenesis. Circulation research 82: 773-778 Weins A, Schwarz K, Faul C, Barisoni L, Linke WA, Mundel P (2001) Differentiation- and stress-dependent nuclear cytoplasmic redistribution of myopodin, a novel actin-bundling protein. The Journal of cell biology 155: 393-404 Witt CC, Burkart C, Labeit D, McNabb M, Wu Y, Granzier H, Labeit S (2006) Nebulin regulates thin filament length, contractility, and Z-disk structure in vivo. The EMBO journal 25: 3843-3855 Wyszynski M, Lin J, Rao A, Nigh E, Beggs AH, Craig AM, Sheng M (1997) Competitive binding of alpha-actinin and calmodulin to the NMDA receptor. Nature 385: 439-442 Zhang Y, Ye J, Chen D, Zhao X, Xiao X, Tai S, Yang W, Zhu D (2006) Differential expression profiling between the relative normal and dystrophic muscle tissues from the same LGMD patient. Journal of translational medicine 4: 53 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/6619 | - |
dc.description.abstract | 我們實驗室在2005年發現了一個基因,命名為nuclear receptor interaction protein (縮寫:NRIP,又名DCAF6或IQWD1)。NRIP的蛋白質結構由七個WD-40 repeats以及一個IQ motif所組成。且實驗室先前的研究證明在有鈣離子的情況下,NRIP會利用IQ motif與調鈣素(Calmodulin)下進行交互作用。利用實驗室的NRIP基因剔除小鼠進行行為測試,其結果暗示NRIP可能參與心臟功能的調控。因此,我們利用心臟超音波檢測,針對NRIP基因剔除小鼠進行長期性的心臟功能監測。根據監測的結果,我們發現剔除NRIP基因確實會影響到小鼠的心臟功能,並且伴隨著年紀增長,心肌有趨於肥厚的現象(cardiac hypertrophy)。為了進一步驗證這項發現,我們將小鼠的心臟取出進行一連串的組織分析,得到的結果與心臟超音波的結果一致。在年紀大的基因剔除小鼠中,我們不但找到心肌肥厚的現象,還發現這些老鼠有心肌纖維化的情形發生。
所以,為了找出NRIP調節心臟功能的分子機制,我們進一步利用yeast two-hybrid進行大量篩選,發現到一群屬於α-actinin家族的蛋白質會與NRIP產生交互作用。α-actinin有四種異構型,其中特定表現在肌肉細胞的異構型ACTN2 是組成肌節上Z-disc的主要成分。Z-disc可以與肌動蛋白絲連結,是維持肌節構形以及穩定肌肉收縮的重要結構。因此,我們利用in vitro及in vivo binding assay進一步確認了NRIP與ACTN2的交互作用,之後又分析兩者間交互作用的區塊並且利用免疫螢光染色法,證明兩者在組織中共同存在於Z-disc上,也意外發現了NRIP是一個新的Z-disc protein。然而,許多論文指出Z-disc protein一旦有缺失便會造成肌節排列錯亂,影響心臟功能而導致心肌症(cardiomyopathy)。因此,我們利用穿透視電子顯微鏡來觀察NRIP基因剔除小鼠的肌節構造。結果顯示缺少NRIP的小鼠其肌節的結構受到影響,特別的是肌節的I-band變窄以及Z-disc變寬。另外,實驗室先前的研究證實NRIP會在有鈣離子的情況下與調鈣素(calmodulin)交互作用,且許多研究指出調鈣素在心肌細胞裡,可以與許多鈣離子通道或者與其他蛋白質交互作用,直接或間接的影響心肌細胞內鈣離子濃度的變化。因此,我們純化並檢測老鼠心肌細胞收縮時鈣離子的變化,發現缺少NRIP會影響心肌細胞收縮時的鈣離子變化量。根據目前的證據我們作出以下的推論:NRIP是一個能夠與ACTN2進行交互作用的Z-disc protein,且缺少NRIP會影響肌節的結構,以及心肌細胞收縮時鈣離的變化量,進而導致心臟收縮功能受損,而引發最終我們看到的心肌肥大的結果。 | zh_TW |
dc.description.abstract | Previously, we demonstrated a novel gene, nuclear receptor interaction protein (NRIP, also named as DCAF6 or IQWD1), which could cooperate with nuclear receptors such as androgen and glucocorticoid receptors and its gene expression was regulated by androgen via androgen receptor. We also identified NRIP as a Ca2+- dependent calmodulin binding protein that activates calcineurin phosphatase activity. To investigate insights into in vivo function of NRIP, we generated NRIP-null mice and found that loss of NRIP impairs cardiac function and lead to cardiac hypertrophy progressively. Furthermore, NRIP-/- mice display weaker muscle strength, reduced cardiac function, and cardiac fibrosis at elder stage compared with WT. To verify the regulatory mechanism, we found that α-actinin-2 (ACTN2), which is a biomarker of muscular Z-disc complex is one of NRIP-interacting proteins from the yeast two-hybrid system. ACTN2 cross-links with actin filament to stabilize sarcomeric structure and muscle contraction, which is an essential constituent of sarcomere. Through the in vitro and in vivo binding assays, we further confirmed the interaction and defined the interacting domains between NRIP and ACTN2. Plus co-localization of NRIP and ACTN2 was discovered in cardiac tissue by immunofluorescence assays, we firstly defined NRIP as a Z-disc protein. Although the Z-disc has been viewed as a passive constituent of the sarcomere traditionally, increasing numbers of mutations in Z-disc proteins leading to disruption and malfunction of the contractile apparatus have been shown to cause cardiomyopathies and/or muscular dystrophies. Hence, we analyzed the sarcomeric structure of NRIP-/- cardiomyocytes and found reduction of I-band width and extension of Z-disc. Besides, we know that NRIP is a Ca2+- dependent calmodulin binding protein. In cardiomyocytes, calmodulin interacts with multiple calcium ion channels or proteins to directly or indirectly regulate the variation of calcium concentration during muscle contraction. Therefore, we isolated and measured the calcium transient of cardiomyocytes. Then, we found that deficiency of NRIP decreases the amplitude of calcium transient. In a conclusion, we speculated that loss of NRIP impairs the structure of sarcomere, the amplitude of calcium transient during muscle contraction and the function of muscle contraction resulting in cardiomyopathy. | en |
dc.description.provenance | Made available in DSpace on 2021-05-17T09:15:21Z (GMT). No. of bitstreams: 1 ntu-101-R99445121-1.pdf: 6387074 bytes, checksum: 80b2d4e34f7864872720eac4fbe4a372 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 致謝 iii
中文摘要 iv ABSTRACT vi CHAPTER 1 INTRODUCTION 1.1 The background knowledge of nuclear receptor interaction protein, NRIP. 1 1.2 Abnormality of calcium homeostasis or sarcomeric proteins leads to cardiomyopathy. 2 1.3 Z-disc protein, sarcomere structure and cardiomyopathy. 3 1.4 The characteristic of ACTN2. 4 1.5 Aims of this study 6 CHAPTER 2 MATRRIALS AND METHODS 2.1 Plasmids and constructs 7 2.2 Cell culture 8 2.3 In vitro Binding Assay 8 2.4 Transfection and Immunoprecipitation assay 9 2.5 Western blot analysis 10 2.6 Histological analysis 11 2.7 Hematoxylin and Eosin (H&E) Staining Protocol 11 2.8 Immunofluorescence assay 11 2.9 Transmission electron microscopy analysis 12 2.10 Antibody 12 2.11 Adult cardiomyocytes isolation 13 CHAPTER 3 RESULTS 3.1 Loss of NRIP leads to cardiac hypertrophy progressively. 15 3.2 NRIP interacts with a Z-disc protein, α-actinin-2, which is a major component of cardiac Z-disc apparatus maintaining the sarcomeric structure. 17 3.3 The IQ motif of NRIP interacts with the CaM-like domain of ACTN2. 18 3.4 NRIP is a novel Z-disc protein and co-localized with ACTN2. 20 3.5 Loss of NRIP reduces I-band length and widen the Z-disc of sarcomere. 21 3.6 Deficiency of NRIP decreases the calcium transient amplitude. 22 CHAPTER 14 DISCUSSION 4.1 Deficiency of NRIP leads to cardiac hypertrophy. 24 4.2 NRIP reduces I-band length through affecting proteins involving in actin filament assembly. 25 4.3 NRIP disrupts myofibrilar arrangements through decreasing gene expressions of genes involving in actin filament formation. 26 4.4 NRIP plays a role in regulating calcium homeostasis. 28 REFERENCE 29 FIGURES 34 APPENDIX 62 | |
dc.language.iso | en | |
dc.title | NRIP缺陷小鼠造成心室肥大 | zh_TW |
dc.title | Deficiency of NRIP causes cardiac hypertrophy | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 顏裕庭,蘇銘嘉,陳佑宗 | |
dc.subject.keyword | NRIP,IQ motif,ACTN2,Z-disc,cardiomyopathy, | zh_TW |
dc.relation.page | 66 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2012-08-10 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 微生物學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
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