本文主要研究内容
作者王宁渤（2019）在《保幼激素通过细胞黏连调节卵泡细胞间通道的分子机制》一文中研究指出：飞蝗(Locusta migratoria)是重要农业害虫,每头雌蝗可产卵300-400粒,强大的生殖力是蝗灾形成的关键因素之一。昆虫的卵黄发生(vitellogenesis)为发育中的卵母细胞提供充足的卵黄蛋白等营养物质,保障了大量的卵成熟和胚胎发育所需的物质和能量。卵黄发生是卵黄原蛋白(vitellogenin,Vg)在脂肪体中合成,通过血淋巴和卵泡细胞间的通道(patency)运输到卵母细胞,最终被卵母细胞吸收储存的过程。在以飞蝗为代表的许多昆虫中,卵黄发生由咽侧体分泌的保幼激素(juvenile hormone,JH)调控。JH诱导卵泡细胞胞间通道开启以促进Vg顺利运输到卵母细胞是昆虫生殖调控的关键进程之一,然而其中的分子机制还有待阐明。本论文利用免疫荧光、Co-IP、Western blot、RNAi等细胞生物学、分子生物学和遗传学的方法,以飞蝗卵泡细胞黏连结构为切入点,鉴定参与JH调节胞间通道的关键细胞黏连因子及信号转导机制,阐明JH通过调控细胞黏连诱导胞间通道开启的分子机制,为害虫绿色防控提供分子靶标。主要研究结果如下:1.飞蝗卵泡细胞以黏着小带(zonula adherens)两两间相互黏附形成紧密的单细胞层包被卵母细胞。通过免疫荧光观察体内黏着小带核心蛋白β-catenin的亚细胞定位发现,随着胞间通道增大,β-catenin定位逐渐收缩。β-catenin仅定位于细胞间黏连的区域,而胞间通道区域无β-catenin定位。体外JH诱导胞间通道开启,发现β-catenin定位也随着胞间通道的开启而变化,与体内实验结果相似。根据上述结果推测,黏着小带在JH诱导的胞间通道开启过程中发生解聚。2.体内注射dsRNA干扰Par3(partitioning-defective protein3)、调节Par3磷酸化的蛋白Par6、或aPKC(atypical protein kinase C),能够破坏卵泡细胞间的黏连,导致卵泡细胞彼此分离,表型与β-catenin RNAi的表型相似。表明Par3、Par6和aPKC不但维系黏着小带结构,也在JH诱导的胞间通道中发挥作用。Co-IP实验表明,JH诱导条件下,β-catenin和Par3的结合显著减弱,暗示JH促进黏着小带的解聚并诱导胞间通道。磷酸化aPKC在卵泡细胞中仅定位在胞间通道区域,而细胞间黏连的区域检测不到磷酸化aPKC信号,推测aPKC参与了胞间通道的调控过程。在体外条件下利用aPKC特异性抑制剂ACPD处理卵黄发生期的卵泡细胞,能够显著抑制JH诱导的胞间通道。此外,aPKC和Par6上游信号因子Cdc42的特异性抑制剂ML141同样能够抑制JH诱导的胞间通道开启。这些结果说明,Cdc42、aPKC、Par3、Par6参与JH诱导的胞间通道。Western blot显示,JH诱导aPKC和Par3磷酸化,而ACPD和ML141抑制JH诱导的aPKC和Par3磷酸化。综合上述结果,JH可能通过Cdc42、Par6、aPKC通路触发Par3磷酸化,磷酸化的Par3与β-catenin分离,导致黏着小带的解聚和胞间通道开启。3.使用G蛋白偶联受体(G protein-coupled receptor,GPCR)抑制剂suramin、受体酪氨酸激酶(receptor tyrosine receptor,RTK)抑制剂genistein和Su6668处理卵黄发生期卵泡细胞并进行JH诱导,发现suramin能够显著抑制JH诱导胞间通道,而genistein和Su6668的处理对胞间通道没有明显影响,表明GPCR参与JH调控的胞间通道。综合本论文的研究结果,我们发现β-catenin主导的黏着小带在JH诱导的胞间通道中起重要作用,JH可能通过GPCR、Cdc42、Par6、aPKC信号通路触发Par3磷酸化,磷酸化的Par3与β-catenin分离,导致黏着小带的解聚,从而诱导胞间通道。研究结果对阐明JH调控卵黄发生的分子机制具有重要意义。
Abstract
fei huang (Locusta migratoria)shi chong yao nong ye hai chong ,mei tou ci huang ke chan luan 300-400li ,jiang da de sheng shi li shi huang zai xing cheng de guan jian yin su zhi yi 。kun chong de luan huang fa sheng (vitellogenesis)wei fa yo zhong de luan mu xi bao di gong chong zu de luan huang dan bai deng ying yang wu zhi ,bao zhang le da liang de luan cheng shou he pei tai fa yo suo xu de wu zhi he neng liang 。luan huang fa sheng shi luan huang yuan dan bai (vitellogenin,Vg)zai zhi fang ti zhong ge cheng ,tong guo xie lin ba he luan pao xi bao jian de tong dao (patency)yun shu dao luan mu xi bao ,zui zhong bei luan mu xi bao xi shou chu cun de guo cheng 。zai yi fei huang wei dai biao de hu duo kun chong zhong ,luan huang fa sheng you yan ce ti fen bi de bao you ji su (juvenile hormone,JH)diao kong 。JHyou dao luan pao xi bao bao jian tong dao kai qi yi cu jin Vgshun li yun shu dao luan mu xi bao shi kun chong sheng shi diao kong de guan jian jin cheng zhi yi ,ran er ji zhong de fen zi ji zhi hai you dai chan ming 。ben lun wen li yong mian yi ying guang 、Co-IP、Western blot、RNAideng xi bao sheng wu xue 、fen zi sheng wu xue he wei chuan xue de fang fa ,yi fei huang luan pao xi bao nian lian jie gou wei qie ru dian ,jian ding can yu JHdiao jie bao jian tong dao de guan jian xi bao nian lian yin zi ji xin hao zhuai dao ji zhi ,chan ming JHtong guo diao kong xi bao nian lian you dao bao jian tong dao kai qi de fen zi ji zhi ,wei hai chong lu se fang kong di gong fen zi ba biao 。zhu yao yan jiu jie guo ru xia :1.fei huang luan pao xi bao yi nian zhao xiao dai (zonula adherens)liang liang jian xiang hu nian fu xing cheng jin mi de chan xi bao ceng bao bei luan mu xi bao 。tong guo mian yi ying guang guan cha ti nei nian zhao xiao dai he xin dan bai β-cateninde ya xi bao ding wei fa xian ,sui zhao bao jian tong dao zeng da ,β-cateninding wei zhu jian shou su 。β-cateninjin ding wei yu xi bao jian nian lian de ou yu ,er bao jian tong dao ou yu mo β-cateninding wei 。ti wai JHyou dao bao jian tong dao kai qi ,fa xian β-cateninding wei ye sui zhao bao jian tong dao de kai qi er bian hua ,yu ti nei shi yan jie guo xiang shi 。gen ju shang shu jie guo tui ce ,nian zhao xiao dai zai JHyou dao de bao jian tong dao kai qi guo cheng zhong fa sheng jie ju 。2.ti nei zhu she dsRNAgan rao Par3(partitioning-defective protein3)、diao jie Par3lin suan hua de dan bai Par6、huo aPKC(atypical protein kinase C),neng gou po huai luan pao xi bao jian de nian lian ,dao zhi luan pao xi bao bi ci fen li ,biao xing yu β-catenin RNAide biao xing xiang shi 。biao ming Par3、Par6he aPKCbu dan wei ji nian zhao xiao dai jie gou ,ye zai JHyou dao de bao jian tong dao zhong fa hui zuo yong 。Co-IPshi yan biao ming ,JHyou dao tiao jian xia ,β-cateninhe Par3de jie ge xian zhe jian ruo ,an shi JHcu jin nian zhao xiao dai de jie ju bing you dao bao jian tong dao 。lin suan hua aPKCzai luan pao xi bao zhong jin ding wei zai bao jian tong dao ou yu ,er xi bao jian nian lian de ou yu jian ce bu dao lin suan hua aPKCxin hao ,tui ce aPKCcan yu le bao jian tong dao de diao kong guo cheng 。zai ti wai tiao jian xia li yong aPKCte yi xing yi zhi ji ACPDchu li luan huang fa sheng ji de luan pao xi bao ,neng gou xian zhe yi zhi JHyou dao de bao jian tong dao 。ci wai ,aPKChe Par6shang you xin hao yin zi Cdc42de te yi xing yi zhi ji ML141tong yang neng gou yi zhi JHyou dao de bao jian tong dao kai qi 。zhe xie jie guo shui ming ,Cdc42、aPKC、Par3、Par6can yu JHyou dao de bao jian tong dao 。Western blotxian shi ,JHyou dao aPKChe Par3lin suan hua ,er ACPDhe ML141yi zhi JHyou dao de aPKChe Par3lin suan hua 。zeng ge shang shu jie guo ,JHke neng tong guo Cdc42、Par6、aPKCtong lu chu fa Par3lin suan hua ,lin suan hua de Par3yu β-cateninfen li ,dao zhi nian zhao xiao dai de jie ju he bao jian tong dao kai qi 。3.shi yong Gdan bai ou lian shou ti (G protein-coupled receptor,GPCR)yi zhi ji suramin、shou ti lao an suan ji mei (receptor tyrosine receptor,RTK)yi zhi ji genisteinhe Su6668chu li luan huang fa sheng ji luan pao xi bao bing jin hang JHyou dao ,fa xian suraminneng gou xian zhe yi zhi JHyou dao bao jian tong dao ,er genisteinhe Su6668de chu li dui bao jian tong dao mei you ming xian ying xiang ,biao ming GPCRcan yu JHdiao kong de bao jian tong dao 。zeng ge ben lun wen de yan jiu jie guo ,wo men fa xian β-cateninzhu dao de nian zhao xiao dai zai JHyou dao de bao jian tong dao zhong qi chong yao zuo yong ,JHke neng tong guo GPCR、Cdc42、Par6、aPKCxin hao tong lu chu fa Par3lin suan hua ,lin suan hua de Par3yu β-cateninfen li ,dao zhi nian zhao xiao dai de jie ju ,cong er you dao bao jian tong dao 。yan jiu jie guo dui chan ming JHdiao kong luan huang fa sheng de fen zi ji zhi ju you chong yao yi yi 。
论文参考文献
论文详细介绍
论文作者分别是来自河南大学的王宁渤，发表于刊物河南大学2019-09-20论文，是一篇关于保幼激素论文,飞蝗论文,卵泡细胞间通道论文,细胞黏连论文,卵黄原蛋白运输论文,河南大学2019-09-20论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自河南大学2019-09-20论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：保幼激素论文; 飞蝗论文; 卵泡细胞间通道论文; 细胞黏连论文; 卵黄原蛋白运输论文; 河南大学2019-09-20论文;