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no-repeat 6px;background-position:center;margin-top:10px;min-width:55px;display:none;} @media (min-width:768px){.gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption .expand_btn{display:none;}} .gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-text{-webkit-flex:9;-ms-flex:9;flex:9;text-align:left;padding:21px 37px 10px 0px;font-size:16px;font-weight:700;} @media (min-width:768px){.gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-text{padding:19px 18px 10px 18px;}} @media (min-width:992px){.gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-text{padding:19px 21px 10px 21px;}} .gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption 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no-repeat 6px;background-position:center;padding:16px 0 16px 0;min-width:62px;max-width:62px;} @media (min-width:768px){.gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-partial-url{padding:16px 0 17px 0;min-width:52px;max-width:52px;}} @media (min-width:992px){.gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-partial-url{padding:15px 0 17px 0;min-width:60px;max-width:60px;}} .gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-partial-url:hover{-webkit-text-decoration:none;text-decoration:none;} .gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption-text{font-size:14px;padding:0px 18px 17px 22px;} @media (min-width:768px){.gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption-text{padding:0px 15px 15px 18px;}} @media (min-width:992px){.gvEcCq .floats-partial-section .floats-partial-footer .floats-partial-caption-text{padding:0px 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href="//www.newsama.com/journals/ijad/" aria-label="International Journal of Alzheimer’s Disease">国际老年痴呆症杂志》上</gydF4y2Baa>/<gydF4y2Baa class="sc-htpNat bUhGXt link sc-eitiEO jXeALb breadCrumb" href="//www.newsama.com/journals/ijad/contents/year/2012/" aria-label="2012">2012年</gydF4y2Baa>/<gydF4y2Baspan class="sc-bhlBdH UVThf">文章</gydF4y2Baspan> </div> <div class="threeColumn__ThreeColumnWrapper-sc-1bf4078-0 dDyxFF threeColumnWrapper"> <div class="threeColumn__LeftWrapper-sc-1bf4078-1 fkVFlF leftWrapper"> <button type="button" class="sc-eKZiaR dzZrgS section__button " id="sectionButton"><span class="sc-htoDjs diutvq icon__dots dots__detector icon-colored _3n-7Ho7DlfjMlZbrRr9Q4a icon-themeable" title="" font-size="22"></span>文章部分</gydF4y2Babutton> <div class="sc-bEjcJn jLUVhb static_contents__section_links links__hidden "> <div id="leftSectionMenu" class="sc-cbkKFq prcY sc-bGbJRg dVCLgr left_section_menu"> <p>在这一页上</gydF4y2Bap> <span class="sc-likbZx bBmZJZ grey" 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<p class="sc-kZmsYB bkiEfx article_type">研究文章|<gydF4y2Ba!-- -->开放获取</gydF4y2Bap> <div class="sc-RcBXQ ztvYg article_citation"> 体积<gydF4y2Ba!-- --> 2012年<gydF4y2Ba!-- --> <span class="sc-fZwumE cUsFHx">|</gydF4y2Baspan>文章的ID<gydF4y2Ba!-- --> 731063年<gydF4y2Ba!-- --> <span class="sc-fZwumE cUsFHx">|</gydF4y2Baspan> <a class="sc-htpNat bUhGXt link sc-fQejPQ duuwge" href="https://doi.org/10.1155/2012/731063" aria-label="Doi-link">https://doi.org/10.1155/2012/731063</gydF4y2Baa> </div> <div class="sc-LKuAh lcOiDF article_full_citation"> <div class="sc-hzNEM gsmOCv isHide"> <p>托马斯·b·谢伊Sangmook Lee<gydF4y2Ba!-- -->,<gydF4y2Ba!-- --> ”<gydF4y2Baspan class="adjust-article-svg-size">Caspase-Mediated截断(τ)强化聚合</gydF4y2Baspan>”,<gydF4y2Ba!-- --><i>国际老年痴呆症杂志》上</gydF4y2Bai>,<gydF4y2Ba!-- --> 卷。<gydF4y2Ba!-- -->2012年<gydF4y2Ba!-- -->,<gydF4y2Ba!-- --> 文章的ID<gydF4y2Ba!-- -->731063年<gydF4y2Ba!-- -->,<gydF4y2Ba!-- --> 7<gydF4y2Ba!-- -->   页面<gydF4y2Ba!-- -->,<gydF4y2Ba!-- --> 2012年<gydF4y2Ba!-- -->。<gydF4y2Ba!-- --> https://doi.org/10.1155/2012/731063</gydF4y2Bap> </div> <button class="sc-hSdWYo etauus sc-iBEsjs kPshFw" color="">显示引用</gydF4y2Babutton> </div> <h1 class="sc-iRbamj jZOxOO sc-iSDuPN kEVvRd article_title adjust-article-svg-size heading-title">Caspase-Mediated截断(τ)强化聚合</gydF4y2Bah1> </div> <div class="sc-jrIrqw eHMFfo article_authors"> <hr class="sc-cEvuZC XkrFe hr__author"> <span class="">Sangmook李<gydF4y2Basup>1</gydF4y2Basup>和</gydF4y2Baspan> <span class=""><b>托马斯·b·谢伊</gydF4y2Bab><a class="sc-htpNat bUhGXt link" href="mailto:" aria-label="Mail Option"><span class="sc-htoDjs kNoiZu sc-hjRWVT irNWJI icon-colored _2zt-1j0LmaF8UjTQAM413G icon-themeable" title="" font-size="16"></span></a><sup>1</gydF4y2Basup></span> <div class="sc-fHxwqH dMuGGA isHide"> <p><sup>1</gydF4y2Basup><span>细胞神经生物学和神经退行性变的研究中心,生物科学,马萨诸塞大学,一所大学大道,洛厄尔,马01854,美国</gydF4y2Baspan></p> </div> <button class="sc-hSdWYo etauus sc-iQtOjA kfDwZY" color="">显示更多</gydF4y2Babutton> </div> <div class="articleContent__EditorWrapper-sc-2yl9jy-3 evMkvU"> <div class="articleContent__TextWrapper-sc-2yl9jy-8 fYrLtk"> <b>学术编辑器:</gydF4y2Bab>弗朗西斯科·潘</gydF4y2Badiv> <div class="articleContent__PublicationTimeLineWrapper-sc-2yl9jy-4 fjviok"> <div class="articleContent__PublcationCol-sc-2yl9jy-5 hgKieC notPublished"> <span class="articleContent__PublicationField-sc-2yl9jy-6 cIribm">收到了</gydF4y2Baspan> <span class="articleContent__PublicationValue-sc-2yl9jy-7 khHUcu">01 2012年6月</gydF4y2Baspan> </div> <div class="articleContent__PublcationCol-sc-2yl9jy-5 hgKieC notPublished"> <span class="articleContent__PublicationField-sc-2yl9jy-6 cIribm">修改后的</gydF4y2Baspan> <span class="articleContent__PublicationValue-sc-2yl9jy-7 khHUcu">2012年7月27日</gydF4y2Baspan> </div> <div class="articleContent__PublcationCol-sc-2yl9jy-5 hgKieC notPublished"> <span class="articleContent__PublicationField-sc-2yl9jy-6 cIribm">接受</gydF4y2Baspan> <span class="articleContent__PublicationValue-sc-2yl9jy-7 khHUcu">2012年7月30日</gydF4y2Baspan> </div> <div class="articleContent__PublcationCol-sc-2yl9jy-5 hgKieC isPublished"> <span class="articleContent__PublicationField-sc-2yl9jy-6 cIribm">发表</gydF4y2Baspan> <span class="articleContent__PublicationValue-sc-2yl9jy-7 khHUcu">04年9月2012年</gydF4y2Baspan> </div> </div> </div> <article class="sc-jtRlXQ gvEcCq article_body "> <div> <div class="xml-content"> <h4 class="header" id="abstract">文摘</gydF4y2Bah4> <p>Caspase-mediated截断(τ)与聚合。我们检查了操纵半胱天冬酶活动的影响细胞内聚集的突变形式的τ(3 po)形成自发的聚集。半胱天冬酶抑制剂治疗Z-VAD-fmk减少N和c端τ截断,但没有显著降低聚合。staurosporine治疗还存在激活,c端增加但不是n端截断和增强聚合。这些发现表明,半胱天冬酶激活是一个潜在的路线,而不是一个必须的起始步骤,在聚合和N - c端截断贡献不同的聚合。</gydF4y2Bap> <span class="end-abs"></span> <hr xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance"> <p>tauopathies病理特征之一是microtubule-associated tau蛋白的聚合。越来越多的证据突出截断的重要性在起始和τ的强化聚合(<gydF4y2Baa href="#B1">1</gydF4y2Baa>- - - - - -<gydF4y2Baa href="#B8">8</gydF4y2Baa>]。τ截断在氨基酸D421已经发现阿尔茨海默病(AD) [<gydF4y2Baa href="#B3">3</gydF4y2Baa>,<gydF4y2Baa href="#B4">4</gydF4y2Baa>,<gydF4y2Baa href="#B9">9</gydF4y2Baa>,<gydF4y2Baa href="#B10">10</gydF4y2Baa>)和其他tauopathies (<gydF4y2Baa href="#B11">11</gydF4y2Baa>]。τ引入了c端截断的构象变化,随着磷酸化,导致聚合(<gydF4y2Baa href="#B12">12</gydF4y2Baa>- - - - - -<gydF4y2Baa href="#B14">14</gydF4y2Baa>]。</gydF4y2Bap> <p>还存在serine-aspartyl蛋白酶通常被认为是激活细胞凋亡过程中,但也可以激活细胞凋亡(<gydF4y2Baa href="#B15">15</gydF4y2Baa>]。在这方面,而半胱天冬酶激活之前和促进的一团形成(<gydF4y2Baa href="#B1">1</gydF4y2Baa>),tangle-bearing神经元可以存活很长时间<gydF4y2Baa href="#B16">16</gydF4y2Baa>,<gydF4y2Baa href="#B17">17</gydF4y2Baa>]。劈理的τD421被建议由caspase-3 [<gydF4y2Baa href="#B3">3</gydF4y2Baa>,<gydF4y2Baa href="#B4">4</gydF4y2Baa>]。相比之下,分析转基因小鼠显示caspase-6,而不是caspase-3,可能在D421截断τ(<gydF4y2Baa href="#B8">8</gydF4y2Baa>]。在老鼠额外的分析表明,半胱天冬酶激活混乱开始可能不是必须的,而是可能代表多个机制之一为τ聚合(<gydF4y2Baa href="#B18">18</gydF4y2Baa>]。在D13截断(τ),这也可以由caspase-6 [<gydF4y2Baa href="#B19">19</gydF4y2Baa>发现,在广告的大脑<gydF4y2Baa href="#B20">20.</gydF4y2Baa>]。是否caspase-6介导这乳沟<gydF4y2Bai>原位</gydF4y2Bai>还不清楚(<gydF4y2Baa href="#B20">20.</gydF4y2Baa>,<gydF4y2Baa href="#B21">21</gydF4y2Baa>]。任何作用的氨基端乳沟聚合仍有待阐明。</gydF4y2Bap> <p>在此,我们提出证据,τ截断D421不一定是由caspase-3。不管是N -还是C -末端截断对聚合至关重要。</gydF4y2Bap> <p>NB2a / d1细胞培养和转染所描述的<gydF4y2Baa href="#B22">22</gydF4y2Baa>)与质粒表达GFP(绿色荧光蛋白)标记的3 POτ,一种变异的人类τ自发聚集,但保留其微管结合能力(f·s .伍特斯博士的礼物、研究所、德国)(<gydF4y2Baa href="#B23">23</gydF4y2Baa>]。介质含有质粒是在文化上12小时,之后改变了媒介和文化孵化一个额外的60小时,以便积累GFP-3PO [<gydF4y2Baa href="#B22">22</gydF4y2Baa>]。还存在激活,细胞收到1<gydF4y2Bai>μ</gydF4y2Bai>M staurosporine(细胞信号,贝弗利,MA)孵化的最后4小时(<gydF4y2Baa href="#B24">24</gydF4y2Baa>]。抑制半胱天冬酶活性,pan-caspase抑制剂,Z-VAD-fmk (Bnzyloxycarbonyl-Val-Ala-Asp(当地)-fluoromethylketone;100年<gydF4y2Bai>μ</gydF4y2Bai>M;Ellisvile Tocris生物科学,MS),添加了12小时直到细胞转染和维护收获后在72小时(<gydF4y2Baa href="#B25">25</gydF4y2Baa>,<gydF4y2Baa href="#B26">26</gydF4y2Baa>]。</gydF4y2Bap> <p>一夜之间,细胞被固定和反应与兔子anti-GFP(表达载体,宏伟的岛,纽约)和小鼠单克隆抗体Tau12(针对aa9-18;EMD微孔,泰梅库拉,CA)、Tau46(针对aa404 - 441;纽约北部的生化,普莱西德湖)[<gydF4y2Baa href="#B27">27</gydF4y2Baa>]或C3(检测在D421τ裂解;圣克鲁斯生物技术、圣克鲁斯,CA) (<gydF4y2Baa href="#B3">3</gydF4y2Baa>][描述<gydF4y2Baa href="#B22">22</gydF4y2Baa>]。</gydF4y2Bap> <p>文化与PBS洗,潜伏在alexa - 488共轭和若丹明红色x共轭二次抗体室温1小时。量化了使用图像<gydF4y2Basvg height="10.6875" id="M1" style="vertical-align:-0.17554pt;width:5.9499998px;" version="1.1" viewbox="0 0 5.9499998 10.6875" width="5.9499998" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,10.6875)"> <g transform="translate(72,-63.45)"> <text transform="matrix(1,0,0,-1,-71.95,63.67)"> <tspan style="font-size: 12.50px; " x="0" y="0"> J</tgydF4y2Baspan> </text> </g> </g> </svg>(<gydF4y2Baa href="http://rsbweb.nih.gov/ij/" target="_blank">http://rsbweb.nih.gov/ij/</gydF4y2Baa>抓取的图像(上)<gydF4y2Baa href="#B22">22</gydF4y2Baa>]。大约100转染细胞(GFP)的存在证实了量化每3个独立的实验条件。immunofluorescent骨料的强度和相邻区域的等效尺寸比较大约40细胞的细胞质每3个独立的实验条件。值给出了平均百分比平均数±标准误差(SEM)。</gydF4y2Bap> <p>细胞被洗两次冰冷的PBS和刮板的10毫米Tris-HCl Triton-X 100年(pH值7.4)包含1%,100毫米氯化钠,EGTA 1毫米,1毫米EDTA、10%甘油、0.1% SDS, 0.5%脱氧胆酸盐,脱氧核糖核酸酶,蛋白酶和磷酸酶抑制剂鸡尾酒(罗氏应用科学,印第安纳波利斯)在4°C。溶菌产物是通过声波降解法其次是离心(<gydF4y2Basvg height="12.8875" id="M2" style="vertical-align:-1.76814pt;width:62.662498px;" version="1.1" viewbox="0 0 62.662498 12.8875" width="62.662498" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,12.8875)"> <g transform="translate(72,-61.69)"> <text transform="matrix(1,0,0,-1,-71.95,63.5)"> <tspan style="font-size: 12.50px; " x="0" y="0"> 1</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="6.2515001" y="0"> 3</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="12.503" y="0"> ,</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="17.704248" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="23.955748" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="30.207249" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="39.234413" y="0"> ×</tgydF4y2Baspan> </text> </g> </g> </svg><i>g</gydF4y2Bai>20分钟)。根据总样本归一化蛋白质和受到SDS-polyacrylamide凝胶电泳和转移到PVDF膜。膜被封锁5% BSA和5毫米的氟化钠TBST (TBS渐变20 0.1%)1小时然后孵化与anti-GFP一夜之间在4°C, Tau46, C3, Tau12 Tau5(这与210 - 241 aa反应;莱斯特博士的礼物粘合剂,西北大学,IL),抗体针对裂解caspase-3(自激活需要乳沟的35 kDa发酵菌激活17 kDa, 19 kDa片段;细胞信号,贝弗利,MA) (<gydF4y2Baa href="#B26">26</gydF4y2Baa>),抗体针对caspase-3衬底,保利ADP-ribose聚合酶(PARP、细胞信号、贝弗利,MA) (<gydF4y2Baa href="#B26">26</gydF4y2Baa>,<gydF4y2Baa href="#B28">28</gydF4y2Baa>]。膜是用相同的缓冲区,孵化与碱性phosphatase-conjugated二级抗体室温1小时,然后使用电视台/ BCIP衬底开发工具包(Promega麦迪逊,WI)。光密度水平的免疫反应性的物种分别量化3免疫印迹,每个由一个独立的实验中,使用图像J。</gydF4y2Bap> <p>微量的裂解caspase-3存在于未经处理的细胞,与基底半胱天冬酶活动的水平一致。Staurosporine caspase-3分开的水平增加了34%,增加已知caspase-3衬底的乳沟,PARP,三倍比未经处理的细胞。Z-VAD-fmk pan-caspase抑制剂,并没有减少稳态乳沟caspase-3和PARP的产物,但阻止完好无损的乳沟PARP 21%,暗示的存在还存在激活前添加抑制剂。Co-incubation staurosporine和Z-VAD-fmk减毒staurosporine-induced半胱天冬酶活动,确认抑制caspase-3活动Z-VAD-fmk(数字<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig1/" target="_blank">1</gydF4y2Baa>(一)和<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig1/" target="_blank">1</gydF4y2Baa>(b))。</gydF4y2Bap> <div class="floats-partial-section partial-carousel-wrapper" data-section="fig1"> <div class="floats-partial-content fig-content-fig1"> <div class="partial-carousel--single-image"> <img alt="" name="731063.fig.001" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.001.jpg"> <br> <strong></strong> </div> </div> <div class="floats-partial-footer"> <div class="floats-partial-caption"> <span class="caption-text">图1</gydF4y2Baspan> <a aria-label="full view" class="caption-partial-url" href="//www.newsama.com/journals/ijad/2012/731063/fig1/" title="" target="_blank"></a> </div> <div class="floats-partial-caption-text"> <svg height="11.25" id="M3" style="vertical-align:-0.30096pt" version="1.1" viewbox="0 0 56.150002 11.25" width="56.150002" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,11.25)"> <g transform="translate(72,-63)"> <text transform="matrix(1,0,0,-1,-71.95,63.35)"> <tspan style="font-size: 12.50px; " x="0" y="0"> </tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="10.890113" y="0"> <</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="22.930502" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="29.182001" y="0"> 。</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="32.307751" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="38.559254" y="0"> 5</tgydF4y2Baspan> </text> </g> </g> </svg> <i>t</gydF4y2Bai> <table> 半胱天冬酶活动的药理操纵。(一)提出了免疫印迹的细胞溶解产物处理Z-VAD-fmk, staurosporine,或者两者兼有,与未经处理的控制,探讨了抗体针对caspase-3裂解,PARP,τ,微管蛋白(如加载控制)表示。相应的图表(b)的平均强度比(±SEM)规范化治疗控制从3免疫印迹。注意增加水平激活caspase-3 PARP乳沟,减少水平的τ后staurosporine治疗。注意Z-VAD-FMK不降低稳态水平caspase-3裂解,裂解PARP,或τ但增加完整的PARP(21%),表明存在持续的半胱天冬酶活性和PARP乳沟前在这些细胞治疗。与Z-VDA-fmk Co-treatment减毒staurosporine-induced半胱天冬酶活动,PARP,τ乳沟。百分比变化被免疫印迹的光密度分析量化。星号显示统计学意义相比,未经处理的控制(,gydF4y2Ba学生的gydF4y2Ba以及)。(c)礼物代表DAPI-stained细胞的图像。注意健康,nonapoptotic原子核,这表明孵化与staurosporine 4小时不诱导细胞凋亡。</tgydF4y2Baable> </div> </div> </div> <p>由staurosporine整体τ水平也有所减少,最广泛的损失的slowest-migrating亚型。Z-VAD-fmk治疗没有显著增加τ水平但防止损耗slowest-migrating亚型,表明他们的损耗是caspase-mediated截断的结果一个或两个末端(数字<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig1/" target="_blank">1</gydF4y2Baa>(一)和<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig1/" target="_blank">1</gydF4y2Baa>(b))。</gydF4y2Bap> <p>与之前的研究一致(<gydF4y2Baa href="#B23">23</gydF4y2Baa>),凋亡的检测并不是总72小时内孵化posttransfection在未经处理的细胞中,也不是治疗可诱发和药理剂(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig1/" target="_blank">1</gydF4y2Baa>(c))。</gydF4y2Bap> <p>我们接下来考虑这些药理治疗如何影响τ乳沟。τ有多个卵裂网站在两末端(<gydF4y2Baa href="#B29">29日</gydF4y2Baa>)(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig2/" target="_blank">2(一个)</gydF4y2Baa>)。因此我们相比水平的GFP (n端)和T46或C3(糖)的中心处Tau5抗原决定基提供索引的截断在终端(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig2/" target="_blank">2 (b)</gydF4y2Baa>)。</gydF4y2Bap> <div class="floats-partial-section partial-carousel-wrapper" data-section="fig2"> <div class="floats-partial-content fig-content-fig2"> <div class="partial-carousel--single-image"> <img alt="(一)”name="731063.fig.002a" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.002a.jpg"> <br> <strong>(一)</gydF4y2Bastrong> </div> <div class="partial-carousel--single-image"> <img alt="(b)”name="731063.fig.002b" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.002b.jpg"> <br> <strong>(b)</gydF4y2Bastrong> </div> </div> <div class="floats-partial-middle"> <div class="floats-partial-middle--thumbs fig-thumbs-fig2"> <button class="partial-carousel-dot" aria-label="(a)"><img alt="(一)”name="731063.fig.002a" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.002a.jpg"><br><strong>(一)</gydF4y2Bastrong></button> <button class="partial-carousel-dot" aria-label="(b)"><img alt="(b)”name="731063.fig.002b" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.002b.jpg"><br><strong>(b)</gydF4y2Bastrong></button> </div> <div class="partial-carousel-next-prev fig-nav-fig2"> <button class="carousel-prev" aria-label="Previous"></button> <button class="carousel-next" aria-label="Next"></button> </div> </div> <div class="floats-partial-footer"> <div class="floats-partial-caption"> <span class="caption-text">图2</gydF4y2Baspan> <a aria-label="full view" class="caption-partial-url" href="//www.newsama.com/journals/ijad/2012/731063/fig2/" title="" target="_blank"></a> </div> <div class="floats-partial-caption-text"> <svg height="11.25" id="M4" style="vertical-align:-0.30096pt" version="1.1" viewbox="0 0 56.150002 11.25" width="56.150002" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,11.25)"> <g transform="translate(72,-63)"> <text transform="matrix(1,0,0,-1,-71.95,63.35)"> <tspan style="font-size: 12.50px; " x="0" y="0"> </tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="10.890113" y="0"> <</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="22.930502" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="29.182001" y="0"> 。</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="32.307751" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="38.559254" y="0"> 5</tgydF4y2Baspan> </text> </g> </g> </svg> <i>t</gydF4y2Bai> <table> τ截断还存在。(一)描述了GFP-tagged 3 POτ。报告的半胱天冬酶乳沟网站广告的大脑和抗体识别的抗原表位用于截断检测在这个研究指出。绿色荧光蛋白标记的3 poτ是描述。Caspase-3据报道在D25和D241裂开τ。Caspase-6据报道在D13裂开τ,D402或D421。(b)礼物代表免疫印迹细胞溶解产物的表达GFP-3PO有或没有治疗staurosporine或Z-VAD-fmk表示。分子量标准的迁移是表示每个污点的左边。相应的图表介绍了意味着强度(±SEM)来自4个独立的免疫印迹。从anti-GFP的相对强度的乐队,C3,和Tau46规范化的Tau5; lower-molecular weight breakdown products were not considered in these densitometric analyses. An asterisk indicates statistical significance (,学生的gydF4y2Ba测试)。</tgydF4y2Baable> </div> </div> </div> <p>氨基端截断监控通过生成GFP / Tau5的比率。这个比例增加Z-DAV-fmk治疗后,表明τ经历在未经处理的细胞稳态caspase-mediated n端乳沟。没有增加,氨基端截断后指出staurosporine治疗(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig2/" target="_blank">2 (b)</gydF4y2Baa>),这表明半胱天冬酶(s)激活短期(4小时)staurosporine治疗不调解n端解理在这些条件下。存在多个低分子量GFP-reactive片段(即。,retaining their full N-terminal region) regardless of manipulation of caspase activity (Figure<a href="//www.newsama.com/journals/ijad/2012/731063/fig2/" target="_blank">2 (b)</gydF4y2Baa>)表明,蛋白酶以外还存在协调进步的c端截断。</gydF4y2Bap> <p>索引的比例提供的c端截断是Tau46 / Tau5和C3 / Tau5。Staurosporine大幅度增加C3免疫反应性,确认τ乳沟D421 [<gydF4y2Baa href="#B3">3</gydF4y2Baa>]。出乎意料,Z-VAD-fmk C3免疫反应性增加。一个可能的解释是,由于Z-VAD-fmk pan-caspase抑制剂,它可能有不同的抑制多个蛋白水解事件,比如截断的τcaspase-6 D402(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig2/" target="_blank">2(一个)</gydF4y2Baa>),这将保护C3 D421乳沟所产生的免疫反应性。第二,非独家的可能性是,乳沟D421不仅仅是由caspase-3。Staurosporine减少Tau46免疫反应性而Z-VAD-fmk增加;这些改变是预期的,因为截断D421消除越远Tau46抗原决定基(见图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig2/" target="_blank">2(一个)</gydF4y2Baa>)。</gydF4y2Bap> <p>来确定是否操纵τ截断聚合的影响,我们量化细胞显示总量的百分比和聚合每个细胞的数量。Staurosporine细胞包含总量的比例增加,聚合每个细胞的数量(数字<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig3/" target="_blank">3(一个)</gydF4y2Baa>,<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig3/" target="_blank">3 (b)</gydF4y2Baa>)。相比之下,Z-VAD-fmk并未显著降低细胞总量的比例,也没有显著降低聚合每个细胞的数量。这些发现表明,半胱天冬酶激活增强聚合,但可能不是必不可少的聚合。</gydF4y2Bap> <div class="floats-partial-section partial-carousel-wrapper" data-section="fig3"> <div class="floats-partial-content fig-content-fig3"> <div class="partial-carousel--single-image"> <img alt="(一)”name="731063.fig.003a" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.003a.jpg"> <br> <strong>(一)</gydF4y2Bastrong> </div> <div class="partial-carousel--single-image"> <img alt="(b)”name="731063.fig.003b" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.003b.jpg"> <br> <strong>(b)</gydF4y2Bastrong> </div> <div class="partial-carousel--single-image"> <img alt="(c)”name="731063.fig.003c" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.003c.jpg"> <br> <strong>(c)</gydF4y2Bastrong> </div> <div class="partial-carousel--single-image"> <img alt="(d)”name="731063.fig.003d" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.003d.jpg"> <br> <strong>(d)</gydF4y2Bastrong> </div> </div> <div class="floats-partial-middle"> <div class="floats-partial-middle--thumbs fig-thumbs-fig3"> <button class="partial-carousel-dot" aria-label="(a)"><img alt="(一)”name="731063.fig.003a" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.003a.jpg"><br><strong>(一)</gydF4y2Bastrong></button> <button class="partial-carousel-dot" aria-label="(b)"><img alt="(b)”name="731063.fig.003b" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.003b.jpg"><br><strong>(b)</gydF4y2Bastrong></button> <button class="partial-carousel-dot" aria-label="(c)"><img alt="(c)”name="731063.fig.003c" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.003c.jpg"><br><strong>(c)</gydF4y2Bastrong></button> <button class="partial-carousel-dot" aria-label="(d)"><img alt="(d)”name="731063.fig.003d" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.003d.jpg"><br><strong>(d)</gydF4y2Bastrong></button> </div> <div class="partial-carousel-next-prev fig-nav-fig3"> <button class="carousel-prev" aria-label="Previous"></button> <button class="carousel-next" aria-label="Next"></button> </div> </div> <div class="floats-partial-footer"> <div class="floats-partial-caption"> <span class="caption-text">图3</gydF4y2Baspan> <a aria-label="full view" class="caption-partial-url" href="//www.newsama.com/journals/ijad/2012/731063/fig3/" title="" target="_blank"></a> </div> <div class="floats-partial-caption-text"> <svg height="11.25" id="M5" style="vertical-align:-0.30096pt" version="1.1" viewbox="0 0 56.150002 11.25" width="56.150002" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,11.25)"> <g transform="translate(72,-63)"> <text transform="matrix(1,0,0,-1,-71.95,63.35)"> <tspan style="font-size: 12.50px; " x="0" y="0"> </tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="10.890113" y="0"> <</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="22.930502" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="29.182001" y="0"> 。</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="32.307751" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="38.559254" y="0"> 5</tgydF4y2Baspan> </text> </g> </g> </svg> <i>t</gydF4y2Bai> <i>μ</gydF4y2Bai> <svg height="11.25" id="M6" style="vertical-align:-0.30096pt" version="1.1" viewbox="0 0 56.150002 11.25" width="56.150002" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,11.25)"> <g transform="translate(72,-63)"> <text transform="matrix(1,0,0,-1,-71.95,63.35)"> <tspan style="font-size: 12.50px; " x="0" y="0"> </tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="10.890113" y="0"> <</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="22.930502" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="29.182001" y="0"> 。</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="32.307751" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="38.559254" y="0"> 5</tgydF4y2Baspan> </text> </g> </g> </svg> <i>t</gydF4y2Bai> <svg height="11.25" id="M7" style="vertical-align:-0.30096pt" version="1.1" viewbox="0 0 56.150002 11.25" width="56.150002" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,11.25)"> <g transform="translate(72,-63)"> <text transform="matrix(1,0,0,-1,-71.95,63.35)"> <tspan style="font-size: 12.50px; " x="0" y="0"> </tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="10.890113" y="0"> <</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="22.930502" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="29.182001" y="0"> 。</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="32.307751" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="38.559254" y="0"> 5</tgydF4y2Baspan> </text> </g> </g> </svg> <i>t</gydF4y2Bai> <table> Z-VAD-fmk和staurosporine聚合和截断的影响。(a)提出了量化的细胞包含总量的百分比与没有检测到GFP药理操纵半胱天冬酶的活动。(b)提出了量化的聚合每个细胞的数量。注意,staurosporine显著增加,而Z-VAD-fmk感应探测,但增高无统计学意义减少,细胞总量的比例以及聚合每个细胞的数量。价值观代表均值±SEM从3独立实验(*,gydF4y2Ba学生的gydF4y2Ba以及)。(c)提出了双immunofluorescent染色anti-GFP和C3(上层图像)或Tau46(较低的图像)和Z-VAD-fmk或staurosporine表示如果不治疗。酒吧是20gydF4y2Bam。箭头表示代表聚集。注意,staurosporine-treated细胞显示更强的C3骨料内免疫反应性和很弱Tau46胞质内的免疫反应性。Staurosporine面板包括从不同的转染细胞突出C3 insets,但不是Tau46或GFP,本地化的外围聚集。(d)提出了量化anti-GFP免疫荧光强度,C3和Tau46骨料和邻在同一细胞胞质τ。由线连接所有酒吧是统计上的不同(gydF4y2Ba;学生的gydF4y2Ba以及);连续线连接值总量,而虚线连接细胞溶质的值。值表示均值±SEM (,gydF4y2Ba学生的gydF4y2Ba以及)。</tgydF4y2Baable> </div> </div> </div> <p>最后,我们比较的相对水平截断τ内聚集与周围的细胞质。因为我们添加Z-VAD-fmk转染和维护,直到收获后不久,聚集在Z-VAD-fmk-treated细胞半胱天冬酶抑制的条件下形成的。自staurosporine添加最后4小时收获之前,聚合已经开始staurosporine治疗之前,让我们来评估半胱天冬酶激活现有总量的影响。GFP和Tau46总量的分布以及邻aggregate-free细胞质中未经处理的细胞之间没有显著性差异和对待Z-VAD-fmk或者staurosporine,确认类似水平的τ包含完整的N和C末端被纳入总量独立的半胱天冬酶处理(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig3/" target="_blank">3 (c)</gydF4y2Baa>)。与免疫印迹分析(图一致<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig2/" target="_blank">2 (b)</gydF4y2Baa>边际C3),免疫反应性治疗和Z-VAD-fmk-treated细胞中发现,骨料之间的均匀分布和周围的细胞质(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig3/" target="_blank">3 (c)</gydF4y2Baa>)。Staurosporine C3免疫反应性显著增加,聚合(图中的多数本地化<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig3/" target="_blank">3 (c)</gydF4y2Baa>)。尤其是C3免疫反应性是沿着外围骨料的突出,而GFP和T46被均匀地分布在整个聚合(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig3/" target="_blank">3 (c)</gydF4y2Baa>)。这些集体immunofluorescent观测表明,τ截断在D421纳入现有的聚合,聚合可以形成在其缺席。</gydF4y2Bap> <p>这里,我们利用绿色荧光蛋白的氨基端标记τ。由于GFP标记本身可以裂解,并可能因此妥协解释,我们利用额外的氨基端特定抗体Tau12(识别的抗原决定基aa9-18)。比较免疫印迹分析和anti-GFP Tau12没有透露anti-GFP特定频带(s)在Tau12墨迹缺席,无论半胱天冬酶活动的操作(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig4/" target="_blank">4(一)</gydF4y2Baa>)。Staurosporine治疗减少τ水平比未经处理的细胞如GFP和Tau12示。治疗Z-VAD-fmk统计增加τ与未经处理的细胞水平的anti-GFP并显示显示趋势意义Tau12 (<gydF4y2Basvg height="11.0375" id="M8" style="vertical-align:-0.17555pt;width:56.150002px;" version="1.1" viewbox="0 0 56.150002 11.0375" width="56.150002" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,11.0375)"> <g transform="translate(72,-63.17)"> <text transform="matrix(1,0,0,-1,-71.95,63.4)"> <tspan style="font-size: 12.50px; " x="0" y="0"> </tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="10.890113" y="0"> =</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="22.930502" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="29.182001" y="0"> 。</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="32.307751" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="38.559254" y="0"> 6</tgydF4y2Baspan> </text> </g> </g> </svg>,学生的<gydF4y2Bai>t</gydF4y2Bai>以及)。Immunofluorescent分析anti-GFP和τ12显示相同的τ分布在聚集和细胞溶质在所有条件下(图<gydF4y2Baa href="//www.newsama.com/journals/ijad/2012/731063/fig4/" target="_blank">4 (b)</gydF4y2Baa>)。这些发现支持的有效性anti-GFPτ的氨基端标记条件下使用。</gydF4y2Bap> <div class="floats-partial-section partial-carousel-wrapper" data-section="fig4"> <div class="floats-partial-content fig-content-fig4"> <div class="partial-carousel--single-image"> <img alt="(一)”name="731063.fig.004a" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.004a.jpg"> <br> <strong>(一)</gydF4y2Bastrong> </div> <div class="partial-carousel--single-image"> <img alt="(b)”name="731063.fig.004b" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.004b.jpg"> <br> <strong>(b)</gydF4y2Bastrong> </div> </div> <div class="floats-partial-middle"> <div class="floats-partial-middle--thumbs fig-thumbs-fig4"> <button class="partial-carousel-dot" aria-label="(a)"><img alt="(一)”name="731063.fig.004a" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.004a.jpg"><br><strong>(一)</gydF4y2Bastrong></button> <button class="partial-carousel-dot" aria-label="(b)"><img alt="(b)”name="731063.fig.004b" src="https://static-01.hindawi.com/articles/ijad/volume-2012/731063/figures/731063.fig.004b.jpg"><br><strong>(b)</gydF4y2Bastrong></button> </div> <div class="partial-carousel-next-prev fig-nav-fig4"> <button class="carousel-prev" aria-label="Previous"></button> <button class="carousel-next" aria-label="Next"></button> </div> </div> <div class="floats-partial-footer"> <div class="floats-partial-caption"> <span class="caption-text">图4</gydF4y2Baspan> <a aria-label="full view" class="caption-partial-url" href="//www.newsama.com/journals/ijad/2012/731063/fig4/" title="" target="_blank"></a> </div> <div class="floats-partial-caption-text"> <svg height="11.0375" id="M9" style="vertical-align:-0.17555pt" version="1.1" viewbox="0 0 63.962502 11.0375" width="63.962502" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,11.0375)"> <g transform="translate(72,-63.17)"> <text transform="matrix(1,0,0,-1,-71.95,63.4)"> <tspan style="font-size: 12.50px; " x="0" y="0"> </tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="10.890113" y="0"> =</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="22.930502" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="29.182001" y="0"> 。</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="32.307751" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="38.559254" y="0"> 6</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="44.810753" y="0"> 2</tgydF4y2Baspan> </text> </g> </g> </svg> <i>t</gydF4y2Bai> <svg height="11.25" id="M10" style="vertical-align:-0.30096pt" version="1.1" viewbox="0 0 56.150002 11.25" width="56.150002" xmlns="http://www.w3.org/2000/svg"> <g transform="matrix(1.25,0,0,-1.25,0,11.25)"> <g transform="translate(72,-63)"> <text transform="matrix(1,0,0,-1,-71.95,63.35)"> <tspan style="font-size: 12.50px; " x="0" y="0"> </tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="10.890113" y="0"> <</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="22.930502" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="29.182001" y="0"> 。</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="32.307751" y="0"> 0</tgydF4y2Baspan> <tspan style="font-size: 12.50px; " x="38.559254" y="0"> 5</tgydF4y2Baspan> </text> </g> </g> </svg> <i>t</gydF4y2Bai> <i>μ</gydF4y2Bai> <table> GFP作为τ的氨基端标记的有效性。(a)礼物代表免疫印迹细胞溶解产物的表达GFP-3PO没有任何治疗和治疗staurosporine或Z-VAD-fmk表示。墨迹图与anti-GFP探测或Tau12 (aa9-18)来显示n端截断的τ药理操纵半胱天冬酶的活动。分子量标准的迁移是表示每个污点的左边。注意没有anti-GFP特定频带(s)在Tau12墨迹缺席。相应的图表介绍了意味着强度(±SEM)来自3免疫印迹。的相对强度的乐队从anti-GFP Tau12规范化,各自的未经处理的控制。Staurosporineτ水平下降GFP和病人的屁股。Z-VAD-fmk统计增加τ水平的anti-GFPτ12并显示意义的趋势(gydF4y2Ba、学生gydF4y2Ba以及)。星号表示统计学差异(gydF4y2Ba;学生的gydF4y2Ba以及)。(b)提出了双immunofluorescent染色的细胞治疗或与Z-VAD-fmk或治疗staurosporine表示。酒吧是20gydF4y2Bam。注意anti-GFP之间的相同colocalization模式和Tau12染色不管药理操纵半胱天冬酶的活动。</tgydF4y2Baable> </div> </div> </div> <p>我们的发现表明还存在一个或多个参与截断τ的末端,但符合其他研究[<gydF4y2Baa href="#B8">8</gydF4y2Baa>),表明其他蛋白酶可能参与进步截断。我们还提供证据表明,截断可能不是必不可少的聚合而是caspase-mediated乳沟D421提高聚合和促进公司的τ总量。这些发现表明,半胱天冬酶激活是一个潜在的路线,而不是一个必须的起始步骤,神经原纤维缠结形成和扩大<gydF4y2Baa href="#B18">18</gydF4y2Baa>,<gydF4y2Baa href="#B30">30.</gydF4y2Baa>]。</gydF4y2Bap> </div> <div class="xml-content"> <h4 class="references" id="references">引用</gydF4y2Bah4> <ol> <li id="B1">a . de Calignon l·m·福克斯,r . 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Bergeron和a·c·勒布朗,”积极caspase-6和caspase-6-cleavedτ在神经纤维网线程,神经炎的斑块,神经原纤维缠结和阿尔茨海默氏症,”<gydF4y2Bai>美国病理学杂志》</gydF4y2Bai>,卷165,不。2、523 - 531年,2004页。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://scholar.google.com/scholar_lookup?title=Active%20caspase-6%20and%20caspase-6-cleaved%20tau%20in%20neuropil%20threads,%20neuritic%20plaques,%20and%20neurofibrillary%20tangles%20of%20Alzheimer's%20disease&author=H. Guo&author=S. Albrecht&author=M. Bourdeau&author=T. Petzke&author=C. Bergeron&author=&author=A. C. LeBlanc&publication_year=2004" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B21">王y s Garg e . m . Mandelkow和e . Mandelkow“τ蛋白水解处理,”<gydF4y2Bai>生化社会事务</gydF4y2Bai>,38卷,不。4、955 - 961年,2010页。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://doi.org/10.1042/BST0380955">出版商的网站</gydF4y2Baa><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Proteolytic%20processing%20of%20tau&author=Y. Wang&author=S. Garg&author=E. M. Mandelkow&author=&author=E. Mandelkow&publication_year=2010" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B22">g·f·s . Lee Hall, t·b·谢伊“τ聚合cdk5和GSK3beta势差现象”,<gydF4y2Bai>阿尔茨海默氏症期刊》上</gydF4y2Bai>,26卷,不。2、355 - 364年,2011页。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://scholar.google.com/scholar_lookup?title=Potentiation%20of%20tau%20aggregation%20by%20cdk5%20and%20GSK3beta&author=S. Lee&author=G. F. Hall&author=&author=T. B. Shea&publication_year=2011" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B23">ai Iliev, s . Ganesan g .短打,f·s .武泰”突破模式的序列在微管绑定重复产生瞬时τ聚合和毒性,”<gydF4y2Bai>生物化学杂志</gydF4y2Bai>,卷281,不。48岁,37195 - 37204年,2006页。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://doi.org/10.1074/jbc.M604863200">出版商的网站</gydF4y2Baa><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Removal%20of%20pattern-breaking%20sequences%20in%20microtubule%20binding%20repeats%20produces%20instantaneous%20tau%20aggregation%20and%20toxicity&author=A. I. Iliev&author=S. Ganesan&author=G. Bunt&author=&author=F. S. Wouters&publication_year=2006" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B24">c . a . Belmokhtar j . Hillion, e . Segal-Bendirdjian”Staurosporine通过caspase-dependent和caspase-independent凋亡机制,“<gydF4y2Bai>致癌基因</gydF4y2Bai>,20卷,不。26日,第3362 - 3354页,2001年。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://doi.org/10.1038/sj.onc.1204436">出版商的网站</gydF4y2Baa><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Staurosporine%20induces%20apoptosis%20through%20both%20caspase-dependent%20and%20caspase-independent%20mechanisms&author=C. A. Belmokhtar&author=J. Híllion&author=&author=E. Segal-Bendirdjian&publication_year=2001" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B25">A·弗雷泽和g·埃文,“杀人执照,”<gydF4y2Bai>细胞</gydF4y2Bai>,卷85,不。6,781 - 784年,1996页。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://doi.org/10.1016/S0092-8674(00)81005-3">出版商的网站</gydF4y2Baa><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=A%20license%20to%20kill&author=A. Fraser &author=G. Evan&publication_year=1996" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B26">d . w·尼科尔森,a .阿里:a .索恩柏瑞et al .,“识别和抑制哺乳动物的细胞凋亡,所需的冰/ CED-3蛋白酶”<gydF4y2Bai>自然</gydF4y2Bai>,卷376,不。6535年,37-43,1995页。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://scholar.google.com/scholar_lookup?title=Identification%20and%20inhibition%20of%20the%20ICE/CED-3%20protease%20necessary%20for%20mammalian%20apoptosis&author=D. W. Nicholson&author=A. Ali&author=N. A. Thornberry et al.&publication_year=1995" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B27">k . s . Kosik l . d . Orecchio l .粘合剂j .问:Trojanowski v . m . y . Lee)和g·李,“跨τ分子的抗原表位配对螺旋细丝,共享”<gydF4y2Bai>神经元</gydF4y2Bai>,1卷,不。9日,第825 - 817页,1988年。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://scholar.google.com/scholar_lookup?title=Epitopes%20that%20span%20the%20tau%20molecule%20are%20shared%20with%20paired%20helical%20filaments&author=K. S. Kosik&author=L. D. Orecchio&author=L. Binder&author=J. Q. Trojanowski&author=V. M. Y. Lee&author=&author=G. Lee&publication_year=1988" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B28">y . a . Lazebnik s h·考夫曼,s . Desnoyers g . g .地方和w·c·恩萧,“乳沟的保利(ADP-ribose)聚合酶蛋白酶与属性如冰,“<gydF4y2Bai>自然</gydF4y2Bai>,卷371,不。6495年,第347 - 346页,1994年。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://doi.org/10.1038/371346a0">出版商的网站</gydF4y2Baa><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Cleavage%20of%20poly(ADP-ribose)%20polymerase%20by%20a%20proteinase%20with%20properties%20like%20ICE&author=Y. A. Lazebnik&author=S. H. Kaufmann&author=S. Desnoyers&author=G. G. Poirier&author=&author=W. C. Earnshaw&publication_year=1994" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B29">r·a·c·w·柯特曼w·w·Poon Rissman,和m . Blurton-Jones“半胱天冬酶的作用乳沟τ的阿尔茨海默病神经病理学,”<gydF4y2Bai>神经病理学和实验神经学杂志》上</gydF4y2Bai>,卷64,不。2、104 - 112年,2005页。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://scholar.google.com/scholar_lookup?title=The%20role%20of%20caspase%20cleavage%20of%20tau%20in%20Alzheimer%20disease%20neuropathology&author=C. W. Cotman&author=W. W. Poon&author=R. A. Rissman&author=&author=M. Blurton-Jones&publication_year=2005" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> <li id="B30">j . h . g . v . w·约翰逊和曹”糖原合成酶激酶3<gydF4y2Bai>β</gydF4y2Bai>诱发caspase-cleavedτ原位聚合。”<gydF4y2Bai>生物化学杂志</gydF4y2Bai>,卷279,不。52岁,54716 - 54723年,2004页。<gydF4y2Baspan class="reflinks">视图:<gydF4y2Baa href="https://doi.org/10.1074/jbc.M403364200">出版商的网站</gydF4y2Baa><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Glycogen%20synthase%20kinase%203β%20induces%20caspase-cleaved%20tau%20aggregation%20in%20situ&author=J. H. Cho &author=G. V. W. Johnson&publication_year=2004" target="_blank">谷歌学术搜索</gydF4y2Baa></span></li> </ol> </div> <div class="xml-content"> <h4 class="copyright" id="copyright">版权</gydF4y2Bah4> <p>版权©2012 Sangmook李和托马斯·b·谢伊。这是一个开放的分布式下文章<gydF4y2Baa rel="license" href="http://creativecommons.org/licenses/by/3.0/">知识共享归属许可</gydF4y2Baa>,它允许无限制的使用、分配和复制在任何媒介,提供最初的工作是正确引用。</gydF4y2Bap> </div> </div> </article> </div> </div> </div> <div class="biblio__Wrapper-sc-1nou1lz-0 eyHfWh journal_article__BiblioIntegratorWidget-sc-12fk6zy-5 dbAANp article_widget biblioWrapper hide"> <hr> <h4 class="biblio__H4-sc-1nou1lz-2 becpDt hrTitleRef">更多相关文章</gydF4y2Bah4> <div id="biblio_widget" class="biblio__BiblioWrapper-sc-1nou1lz-1 hFGNjP"></div> </div> </div> </div> <div class="threeColumn__RightWrapper-sc-1bf4078-2 jzTMLK rightWrapper" id="rightSectionMenu"> <div> <div class="journal_article__ArticleStickyTopToolbar-sc-12fk6zy-0 gbrQxi article_sticky_top_toolbar"> <div class="sc-cBdUnI hRnXob article_top_toolbar"> <a class="sc-htpNat bUhGXt link sc-jtRfpW dIjoCg sc-exkUMo hymroZ pdf_desktop button__group" href="https://downloads.hindawi.com/journals/ijad/2012/731063.pdf" aria-label="" target="_blank" id="toolbar__dcNavSmall"><span><img alt="" class="sc-EHOje jOLhQl sc-kTUwUJ eujois" title="" role="presentation" src="https://static-01.hindawi.com/next_assets/wyT4heYybLmt1UEJexPE9/_next/static/node_modules/hindawi-ui/src/icon/svg/pdf.svg" height="24"><span class="text">PDF</gydF4y2Baspan></span><span class="sc-htoDjs bqeMOy sc-dqBHgY cQgXqt fileArrow icon-colored _3P4iUn1NDM5G77nWCamJ4l icon-themeable" title="" font-size="20"></span></a> <a class="sc-htpNat bUhGXt link sc-jtRfpW dIjoCg sc-kcDeIU cbcyj ref_desktop" href="//www.newsama.com/journals/ijad/2012/731063.ris" aria-label="" target="" id="toolbar__dcNavLarge"><span><img alt="" class="sc-EHOje jOLhQl sc-kTUwUJ eujois" title="" role="presentation" 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height="24"><span class="text">下载引用</gydF4y2Baspan></span><span class="sc-htoDjs bqeMOy sc-dqBHgY cQgXqt fileArrow icon-colored _3P4iUn1NDM5G77nWCamJ4l icon-themeable" title="" font-size="20"></span></a> <a class="sc-htpNat bUhGXt link sc-jtRfpW dIjoCg sc-BngTV hmDWcT ref_desktop" href="//www.newsama.com/journals/ijad/2012/731063.ris" aria-label="" target="" id="toolbar__dcNavSmall"><span><img alt="" class="sc-EHOje jOLhQl sc-kTUwUJ eujois" title="" role="presentation" src="data:image/svg+xml;base64,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" height="24"><span class="text">引用</gydF4y2Baspan></span><span class="sc-htoDjs bqeMOy sc-dqBHgY cQgXqt fileArrow icon-colored _3P4iUn1NDM5G77nWCamJ4l icon-themeable" title="" font-size="20"></span></a> </div> </div> <div class="journal_article__ArticleStickyBottomToolbar-sc-12fk6zy-1 isgRZA article_sticky_bottom_toolbar"> <div class="sc-cTjmhe ihmyEq article_bottom_toolbar"> <div class="sc-cugefK itFKcQ accordion" property="background-color" color="#D8D8D8" id="toolbar__dofNav"> <span><img alt="" class="sc-EHOje jOLhQl sc-bfYoXt cGuwDn" title="" role="presentation" src="data:image/svg+xml;base64,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" height="24"><span class="sc-dRFtgE jrsDZP">下载其他格式</gydF4y2Baspan><span class="sc-gkFcWv gWTCXQ">更多的</gydF4y2Baspan></span> <span class="sc-htoDjs bMdQcY sc-gbOuXE jGPvPL icon-colored _1CwL9R82TPl2HxDleeYGDR icon-themeable" title="" font-size="11"></span> </div> <div class="sc-fnwBNb etzuBr"> <a class="sc-htpNat bUhGXt link sc-iNhVCk gWgCNb open_panel" href="//www.newsama.com/downloads/journals/ijad/2012/731063.epub" aria-label="ePub" property="background-color" color="#D8D8D8"> <div class="sc-eAKXzc WHkfb"> ePub</gydF4y2Badiv><img alt="" class="sc-EHOje jOLhQl sc-bfYoXt cGuwDn" title="" role="presentation" src="data:image/svg+xml;base64,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" height="24"></a> <a class="sc-htpNat bUhGXt link sc-iNhVCk gWgCNb open_panel" href="//www.newsama.com/downloads/journals/ijad/2012/731063.xml" aria-label="XML" property="background-color" color="#D8D8D8"> <div class="sc-eAKXzc WHkfb"> XML</gydF4y2Badiv><img alt="" class="sc-EHOje jOLhQl sc-bfYoXt cGuwDn" title="" role="presentation" 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height="24"><span class="sc-dRFtgE jrsDZP">订单打印副本</gydF4y2Baspan><span class="sc-gkFcWv gWTCXQ">订单</gydF4y2Baspan></span><span class="sc-htoDjs kNoiZu sc-gbOuXE jGPvPL right__icon icon-colored _32KP5uexH-Bgs7Kj8Gk1Tz icon-themeable" title="" font-size="16"></span></a> <div class="sc-hBbWxd kdRRST"> <div class="sc-dyGzUR cHatnS"> <div> 的观点<gydF4y2Baspan>2351年</gydF4y2Baspan> </div> <div> 下载<gydF4y2Baspan>1111年</gydF4y2Baspan> </div> </div> <div> <div class="sc-drKuOJ kxdGAZ"> <span class="__dimensions_badge_embed__" data-doi="10.1155/2012/731063" data-style="small_circle"></span>引用</gydF4y2Badiv> </div> </div> <nav class="sc-bRBYWo fXBKFa sc-hUfwpO jHPgUP" id="toolbar__iconNav"> <h3 class="sc-Rmtcm hNPrIr">分享文章</gydF4y2Bah3> <ul class="sc-hzDkRC bnbfTS"> <li class="iconBorder"><a class="sc-htpNat bUhGXt link" href="https://twitter.com/share?url=https://doi.org/10.1155/2012/731063&via=Hindawi" aria-label="Sharing on Twitter" rel="noopener noreferrer" target="_blank"><img alt="" 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