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Time course of expression of intermediate filament protein vimentin,nestin and desmin in rat renal glomerular injury

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Chinese Medical Journal 2007年第13期120卷 1203-1205页

作者： ZOU Jun CHANG Tian-hui CHANG He Eishin Yaoita Yutaka Yoshida Masaaki Nameta Tadashi Yamamoto JIN Xin Department of Pharmacy School of Medical Sciences Xiamen University Xiamen 361005 China Department of Clinical Pharmacology School of Pharmaceutical Sciences China Medical University Shengyang 110001 China Division of Cardiology Niigata University Graduate School of Medical and Dental Sciences 1-757 Asahimachi-dori Niigata 951-8510 Japan Department of Structural Pathology Institute of NephrologyNiigata University Graduate School of Medical and Dental Sciences 1-757 Asahimachi-dori Niigata 951-8510 Japan

Podocytes in renal glomemlus express unusual intermediate filament proteins （IFs） for visceral epithelial cells. IFs cytoskeleton is mainly composed of vimentin, nestin and desmin. Tissue injury is often accompanied by changes in gene expression of IFs. Enhanced desmin staining in variety of rat experimental including 2 4 puromycin nephrosis. It has not podocytes are observed in a models of podocyte injury aminonucleoside （PAN） been elucidated whether expression of vimentin and nestin is up-regulated in podocyte injury. To further gain insight into expression of IFs in podocytes, we investigated the time course of vimentin, nestin and desmin in PAN nephrosis.

关键词： intermediate filament vimentin nestin desmin kidney

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Mechanical properties of crosslinks controls failure mechanism of hierarchical intermediate filament networks

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Theoretical & Applied Mechanics Letters 2012年第1期2卷 27-31页

作者： Zhao Qin Markus J. Buehler Laboratory for Atomistic and Molecular Mechanics （LAMM） Department of Civil and Environmental Engineering Massachusetts Institute of Technology 77 Massachusetts Ave. Room 1-235 A＆B Cambridge MA 02139 USA Center for Computational Engineering Massachusetts Institute of Technology 77 Massachusetts Ave. Cambridge MA 02139 USA Center for Materials Science and Engineering Massachusetts Institute of Technology 77 Massachusetts Ave. Cambridge MA 02139 USA

intermediate filaments are one of the key components of the cytoskeleton in eukaryotic cells, and their mechanical properties are found to be equally important for physiological function and disease. While the mechanical properties of single full length filaments have been studied, how the mechanical properties of crosslinks affect the mechanical property of the intermediate filament network is not well understood. This paper applies a mesoscopic model of the intermediate network with varied crosslink strengths to investigate its failure mechanism under the extreme mechanical loading. It finds that relatively weaker crosslinks lead to a more flaw tolerant intermediate filament network that is also 23% stronger than the one with strong crosslinks. These findings suggest that the mechanical properties of interfacial components are critical for bioinspired designs which provide intriguing mechanical properties.

关键词： failure mechanism flow tolerance intermediate filament protein network soft material,rupture crosslink strength bioinspired design

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Effect of dl-praeruptorin A on desmin and vimentin content in rat ischemia/reperfusion myocardiocytes

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Chinese Medical Journal 2007年第24期120卷 2256-2259页

作者： CHANG He CHU Xiao-yu ZOU Jun CHANG Tian-hui Division of Cardiology Graduate School of Medical and DentalSciences Niigata University 1-757 Asahimachi-dori Niigata951-8510 Japan Department of Clinical Pharmacology School of PharmaceuticalSciences China Medical University Shengyang 110001 China Department of Pharmacy School of Medical Sciences Xiamen University Xiamen 361005 China

Background intermediate filament （IF） proteins have been thought to play a role in nuclear centration, organelle movement and maintenance of cell shape, dl-praeruptorin A （Pd-la）, a novel Ca^2＋-influx blocker and K^＋-channel opener isolated from Chinese traditional herbal medicine Qian-Hu, has been demonstrated to inhibit expression of apoptosis related proteins and reduce the level of proinflammatory factors in ischemia/reperfusion myocardiocytes. Morphologically, whether Pd-la effects myocardiocyte IFs remains unclear. The purpose of this study was, for the first time, to evaluate the in vivo effects of Pd-la on IF desmin and vimentin content in order to further explore its cardioprotection against ischemia and elucidate its mechanism of action. Methods Rats underwent a 30 minutes coronary occlusion followed by 120 minutes reperfusion. Assessment of desmin and vimentin content of myocardiocytes was performed by immunohistochemistry, Western blot, Hematoxylin-Eosin staining and densitometry. Results Pretreatment with i.v. infusion of Pd-la prior to ischemia significantly increased desmin and vimentin content and alleviated defects caused by the ischemia/reperfusion insult, e.g. with Pd-la at a dose of 0.5 or 1.0 mg/kg, integrated density values of desmin were increased from 61 478 ± 10 074 to 177 408 ±10 395 and 195 784±20 057, and vimentin from 59 189± 19 853 to 164 781± 19 543 and 185 696± 20 957 （P〈0.01, vs placebo）, respectively. The recovery of desmin seemed to be stronger and appeared earlier than that of vimentin. Conclusion Pd-la protectively increased IF desmin and vimentin content in ischemia/reperfusion myocardiocytes, which might be partially responsible for its cardioprotection against ischemia.

关键词： dl-praeruptorin A desmin vimentin intermediate filament ischemia myocardiocyte

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瘤背石磺内参基因筛选和皮肤发育相关基因EGF和IF的克隆与表达分析

瘤背石磺内参基因筛选和皮肤发育相关基因EGF和IF的克隆与表达分析

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作者：杨铁柱上海海洋大学

学位级别：硕士

瘤背石磺(Onchidium struma)是生活于滩涂湿地的半咸水两栖性生物,隶属于软体动物门(Mollusca)、腹足纲(Gastropoda)、肺螺亚纲(Pulmonata)、缩眼目(Systellommatophora)、石磺科(Onchidiidae),是一种具有很高的食用和药用价值的贝类。其生活区域常见于中国的江苏省、上海市、浙江省、福建省、广东省等地的淡水与海水交汇处的高潮带,其温度适应能力强、分布范围广、野生资源量非常丰富。目前国内外学者对于石磺的研究工作主要集中在系统分类学、生物学形态特征、营养价值评定、胚胎发育和繁殖机制以及活性物质的分离提取等方面。表皮生长因子可促进上皮细胞的增殖发育,中间丝蛋白在维持细胞形态上发挥重要作用,内参基因的筛选是进行基因定量分析的前提条件。石磺科的蜕皮现象涉及到上皮细胞的大量更新,其运动能力则与肌肉细胞的形变程度有联系。本文对瘤背石磺进行了内参基因的初步筛选,初步探讨功能基因EGF和IF的理化结构和进化关系,并针对两基因在石磺蜕皮与否和运动能力强弱上进行了较为深入的分析,希望能为研究瘤背石磺生物学类两栖特性相关的功能基因积累资料。1.依托瘤背石磺转录组分析数据库,挑选出7个常用内参基因的序列作为候选基因,经NCBI上BLAST比对及初步鉴定后,设计荧光定量PCR引物,统计分析各候选基因在瘤背石磺不同组织中表达的稳定性。BestKeeper软件分析显示其稳定性大小为:EF1?>TUBB>RPL28>Ubiq>18S rRNA=CYC>ACTB;在geNorm软件的分析结果里,稳定性的大小为:EF1?>RPL28>CYC>Ubiq>TUBB>18S rRNA>ACTB;而Normfinder软件给出的稳定值排序为:EF1?>RPL28>Ubiq>CYC>TUBB>18S RNA>ACTB。常用内参基因的ACTB的标准差(SD)最大,其表达稳定性在7个候选基因中最低,稳定性最高的候选基因为EF1?。在瘤背石磺的不同组织表达分析时筛选出最适合内参基因为EF1?,而由Normfinder软件给出的最优内参基因组合为EF1?和RPL28。2.从瘤背石磺背部皮肤组织中克隆得到了瘤背石磺表皮生长因子(Os-egf1)基因,并进行序列分析、结构预测和组织表达水平测定。利用RACE克隆技术所获得的Os-egf1基因的cDNA序列全长为1158bp,其开放阅读框(ORF)长度为846bp,并编码一条长为281个氨基酸残基的多肽链;没有明显的信号肽,分子量为30.51kDa,该基因存在一个跨膜结构域(膜内1-16aa,膜上17-39aa,膜外40-281aa),亚细胞定位发现该多肽位于细胞核内,理论等电点(PI)是5.41,为一个偏酸性的蛋白。有6个Ser、3个Thr、3个Tyr可能为蛋白激酶磷酸化作用的位点。Os-egf1氨基酸的序列中半胱氨酸(Cys)残基明显较多,占氨基酸残基总数的11.40%,并不分布于多肽链的N端和C端,多集中于肽链的中段部分。半胱氨酸残基主要分布在对应的三个EGF-like的结构域里,且结构域中至少6个半胱氨酸残基参与形成CX7 CX4-5 CX10-13 CXCX8 C的类似结构,推测其相互之间形成的3对二硫键为维持结构域特定的空间结构所必需。除去结构域中的半胱氨酸残基,该基因的序列保守性非常低,进化树结果显示EGF家族中不同成员各自聚为一支。瘤背石磺不同组织的相对定量分析显示Os-egf1高表达于背部皮肤,表皮生长因子可促进上皮细胞的增殖分裂,初步推测Os-egf1基因与瘤背石磺蜕皮现象明显与否有关。3.从瘤背石磺背部皮肤组织中克隆了瘤背石磺中间丝蛋白(Os-IF1)基因,序列拼接结果显示,所克隆得到的Os-IF1基因的cDNA核酸序列全长为1957bp,其开放阅读框长度为1359bp,并编码一条长为452个氨基酸残基的多肽链。没有明显的信号肽,分子量为51.54kDa,不存在跨膜结构域,亚细胞定位发现该多肽位于细胞核内,理论等电点为5.21,为一个偏酸性的蛋白,含有23个Ser、14个Thr、6个Tyr蛋白激酶磷酸化位点。Os-IF1预测的氨基酸序列中,以?螺旋为主,占据整条多肽链的82%,符合中间丝蛋白的典型特征。氨基酸序列的同源性比对发现其与中间丝蛋白家族成员的序列同源性较高。在相对定量表达分析中,Os-IF1在瘤背石磺中表达量较高的为前触角和腹足两个组织,而其他三种石磺表达量最高的组织均为腹足,推测该基因产物与石磺运动过程中腹足肌肉的收缩有关。不同种石磺之间的表达分析显示,瘤背石磺中的表达量最高,具体原因有待于进一步的探究。序列比对中发现该基因的保守性非常高,而进化树基本表现为:腹足纲、头足纲、双壳纲、脊索动物和脊椎生物各自聚在一起形成分支,与瘤背石磺进化关系最

关键词：瘤背石磺内参基因 EGF intermediate filament 系统进化荧光定量表达

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Nestin is a critical prosurvival factor in neural stem/progenitor cells

Nestin is a critical prosurvival factor in neural stem/proge...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Donghyun Park Andy Peng Xiang Frank Fuxiang Mao Noah Walton Bruce T.Lahn Howard Hughes Medical Institute Department of Human GeneticsUniversity of ChicagoChicago IL 60637USA Center for Stem Cell Biology and Tissue Engineering Sun Yat-sen University

＜正＞The intermediate filament protein,nestin,is a widely employed marker of multipotent neural stem/progenitor cells（NSCs）.Recent in vitro studies have implicated nestin in a number of cellular processes,but there is no evidence yet on its in vivo ***,we report the construction and functional characterization of Nestin knockout *** found mat these mice show embryonic lethality,with neuroepithelia of the developing neural tube exhibiting significantly increased *** with this in vivo observation,NSC cultures derived from knockout embryos show dramatically elevated apoptosis but no defects in proliferation or ***,nestin deficiency has no deflectable effect on the integrity of the ***,the knockout of vimentin,while abolishing nestin’s ability to polymerize into intermediate filaments in NSCs,does not lead to any apoptotic *** data demonstrate that nestin is a critical prosurvival factor in NSCs,and that nestin’s prosurvival function is surprisingly uncoupled from its structural involvement in the cytoskeleton.

关键词： nestin intermediate filament apoptosis neural stem cells neural progenitor cells vimentin

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IF-Like Protein Ngs is Essential for Maintenance of Chordocytes in Zebrafish

IF-Like Protein Ngs is Essential for Maintenance of Chordocy...

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“基因、进化与生理功能多样性”海内外学术研讨会暨中国生理学会第七届比较生理学学术会议

作者：夏梽丹 Helena Telfer Key Laboratory of Cell Proliferation and Differentiation Center of Developmental Biology and GeneticsCollege of Life SciencesPeking UniversityMinistry of Education University of Washington

The notochord is a characteristic structure for all members of the phylum Chordata, which plays critical roles in embryonic development,patterning and locomotion in *** screening a pool of retroviral insertions,we isolated a mutant zebrafish (Danio rerio) line which shows defective notochord with a granular surface when bred to *** phenotype was found to be caused by the retroviral insertion in a novel gene encoding an intermediate filament(IF) like *** therefore named the mutant and the related gene as"ngs"(notochord granular surface).Whole mount in situ hybridization revealed that the gene is specifically expressed in notochord,and knocking down of ngs by morpholino injection produced similar notochord *** observation demonstrated that the granular surface is due to aggregation of small cells originating from notochord epithelial *** electron micrographs of transverse sections of 48hpf(hours post-fertilization) embryos showed that the mutant lacks 10nm filamentlike structures in the notochord cells,while there are plenty of IF like filaments in wild type *** results indicate that ngs gene plays an important role in maintaining notochord structure during zebrafish larval swimming,which reveals a novel function of intermediate filament family in providing crucial structural support for cells.

关键词： Zebrafish Notochord intermediate filament Insertional mutant

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