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Revealing the F_actin Networks in Interphase Nuclei of Garlic Clove Cells by Confocal Fluorescence Microscopy 被引量:2
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作者 王冬梅 王学臣 张伟成 《Acta Botanica Sinica》 CSCD 2000年第11期1167-1171,共5页
The interphase nuclei of parenchyma cells and epidermal cells of garlic ( Allium sativum L.) clove were labelled with rabbit anti_actin antibody and FITC_conjugated goat anti_rabbit IgG antibody. The authors observ... The interphase nuclei of parenchyma cells and epidermal cells of garlic ( Allium sativum L.) clove were labelled with rabbit anti_actin antibody and FITC_conjugated goat anti_rabbit IgG antibody. The authors observed results with fluorescence microscopy and confocal laser scanning microscopy. The nuclei showed prominent green_yellow fluorescence, indicating the presence of actin in the nuclei. Fluorescence examination with TRITC_phalloidin showed distinctive red fluorescence in the nuclei, indicating that F_actin is present in the nuclei. Confocal laser scanning microscopy indicated the presence of F_actin containing network structures in the nuclei, but the network structures were absent and the nuclei still showed red fluorescence when the cells were treated with cytochalasin D before fixation; the red fluorescence in the nuclei was hard to be observed when the cells were treated with unlabelled phalloidin before the cells were stained with TRITC_phalloidin. These results indicate that F_actin is in the nuclei and forms network structures in the nuclei of garlic cells. 展开更多
关键词 interphase nucleus f_actin TRITC_phalloidin cytochalasin D confocal laser scanning microscopy Allium sativum
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Evidence of Ultrastructure and Physiology of F-actin as Component of Plasmodesmata 被引量:2
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作者 王冬梅 王学臣 张伟成 《Acta Botanica Sinica》 CSCD 2002年第11期1278-1285,共8页
The characters and ultrastructure of the intercellular connection were revealed in the outer epidermis of the garlic clove sheath by means of fluorescent probe TRITC_Phalloidin (TRITC_Ph) labeling combined with confoc... The characters and ultrastructure of the intercellular connection were revealed in the outer epidermis of the garlic clove sheath by means of fluorescent probe TRITC_Phalloidin (TRITC_Ph) labeling combined with confocal laser scanning microscopy (CLSM), immuno_gold labeling and transmission electron microscopy. These results show that transcellular channel is a complex of rod_like cytoplasm channel and grouped plasmodesmata (PDs) in pit. The former remains a portion of the cell protoplast. The diameter of PD is normally 60-70 nm. The PDs are the real intercellular symplasmic connections of the cells. The transcellular fibers labeled with the TRITC_Ph obviously become narrow in the primary pit fields, which is the same as the characters observed under the electron microscope. The bright fluorescent spot in the primary wall reflects the grouped PDs in pit, and hence the presence of F_actin in the PDs can be confirmed. In immuno_gold labeling experiment, a lot of gold particles were massively distributed in the rod_like cytoplasm channel and grouped PDs. The result provides effective support that these fluorescent filaments possibly are the existing form of F_actin. 展开更多
关键词 PLASMODESMATA f_actin confocal laser scanning microscopy (CLSM) ULTRASTRUCTURE Allium sativum
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Identification of Myosinon the Surface of Wheat Mitochondria
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作者 赵和平 刘爱校 +2 位作者 任东涛 刘国琴 阎隆飞 《Acta Botanica Sinica》 CSCD 1999年第12期1303-1306,共4页
Itwasdemonstratedthatmyosinwasassociatedwiththesurfaceofmitochondriainwheat (Triticum aestivumL .) .AssaysofpolyacrylamidegelelectrophoresisandWesternblottinghaveshownthatapolypeptide withmolecularweightof2 10kDcoul... Itwasdemonstratedthatmyosinwasassociatedwiththesurfaceofmitochondriainwheat (Triticum aestivumL .) .AssaysofpolyacrylamidegelelectrophoresisandWesternblottinghaveshownthatapolypeptide withmolecularweightof2 10kDcouldberecognizedbyapolyclonalantibodyagainsthumanmusclemyosin .It wasfoundthattheATPaseactivityofmitochondrialsuspensioncouldbestimulatedbyF_actinisolatedfrom chickenmuscle ,whichindicatedthattherewasmyosinonthesurfaceofwheatmitochondria .Thisresultwas confirmedbyelectronmicroscopicobservation :mitochondriatreatedwithN_ethylmaleimide (NEM)couldbe wrappedbytheF_actin . 展开更多
关键词 Mitochondriainwheat MYOSIN f_actin ATPASE LOCALIZATION
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肝癌细胞纤维肌动蛋白的共聚焦激光扫描显微镜光学切片观察 被引量:6
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作者 霍霞 徐锡金 +2 位作者 陈耀文 林珏龙 许险峰 《汕头大学医学院学报》 2001年第3期174-176,F003,共4页
目的 :建立培养细胞共聚焦激光扫描显微镜光学切片的方法 ,探讨肝癌细胞纤维肌动蛋白 (F_actin)的空间结构。方法 :采用共聚焦激光扫描显微镜光学切片技术结合异硫酸氢荧光素_鬼笔环肽 (FITC_phalloidin)标记纤维肌动蛋白和碘化丙啶(PI... 目的 :建立培养细胞共聚焦激光扫描显微镜光学切片的方法 ,探讨肝癌细胞纤维肌动蛋白 (F_actin)的空间结构。方法 :采用共聚焦激光扫描显微镜光学切片技术结合异硫酸氢荧光素_鬼笔环肽 (FITC_phalloidin)标记纤维肌动蛋白和碘化丙啶(PI)标记细胞核的荧光探针双重标记技术对肝癌细胞纤维肌动蛋白进行形态学观察和图像分析。结果 :肝癌细胞内纤维肌动蛋白微丝形成束状纤维 ,粗壮而密集 ,平行排列或纵横交错成网状贯穿整个细胞和细胞突起。结论 :① 共聚焦激光扫描显微镜的光学切片技术结合FITC -phalloidin和PI荧光探针双重标记技术是观察和分析细胞骨架系统三维立体结构的良好方法 ;② 肝癌细胞内纤维肌动蛋白微丝粗壮。 展开更多
关键词 肝癌细胞株 纤维肌动蛋白 共聚焦激光扫描显微镜 光学切片
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蒜瓣鞘表皮组织中肌动蛋白纤丝跨胞分布的共焦荧光显微镜观察 被引量:3
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作者 王冬梅 王学臣 张伟成 《Acta Botanica Sinica》 CSCD 2000年第3期327-330,共4页
By means of paraformaldehyde fixation, Triton X100 extraction and TRITCphalloidin staining, the presence and distribution patterns of Factin in the outer epidermal cells of the garlic (Allium sativum L.) sheath were... By means of paraformaldehyde fixation, Triton X100 extraction and TRITCphalloidin staining, the presence and distribution patterns of Factin in the outer epidermal cells of the garlic (Allium sativum L.) sheath were studied with fluorescence probe technique and confocal laser scanning microscopy. There were a lot of actin filaments (AFs) impenetrate the cell wall, but the AFs with red fluorescence were absent when the cells were treated with cytochalasin D before fixation; the same result was obtained when the cells were treated with unlabeled phalloidin. These results indicate the presence of Factin in the intercellular channels and that it is related to the plasmodesmata and intercellular trafficking of macromolecules. 展开更多
关键词 胞间连丝 F-肌动蛋白 表皮组织 蒜瓣鞘 显微镜
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成骨样细胞受力后细胞骨架中微丝形态结构变化的初步研究 被引量:5
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作者 陈国平 周征 +1 位作者 郑翼 罗颂椒 《华西口腔医学杂志》 CAS CSCD 北大核心 2002年第3期212-215,共4页
目的 :探讨成骨样细胞株UMR_10 6受机械力作用后细胞骨架微丝蛋白F_actin的变化。方法 :成骨样细胞株UMR_10 6体外扩增 ,实验组以 2 2 5×g相对离心力离心 10min后 ,分 7组分别继续培养 15、30min ,1、4、6、12、2 4h ,并设立相应... 目的 :探讨成骨样细胞株UMR_10 6受机械力作用后细胞骨架微丝蛋白F_actin的变化。方法 :成骨样细胞株UMR_10 6体外扩增 ,实验组以 2 2 5×g相对离心力离心 10min后 ,分 7组分别继续培养 15、30min ,1、4、6、12、2 4h ,并设立相应对照组。所有样本经BODIPYFLPhallacidin处理后 ,在激光扫描共聚焦显微镜下观察、记录图片并测定单个细胞的平均荧光强度。结果 :对照组F_actin较粗 ,呈束状 ,排列较整齐 ;实验组除 2 4h外 ,其余各组的纤维明显较对照组纤细、短 ,散乱分布于胞浆内、无方向性 ;但两组间差异随时间延长逐渐减小。除 2 4h外 ,实验组的平均荧光强度较对照组明显降低 ,差异有显著性 ;随时间延长 ,实验组荧光强度逐渐回升 ,到 2 4h组时较对照组增强。结论 :成骨样细胞受力后细胞骨架微丝的结构明显变化 ,荧光染色变弱 ,表明在机械力作用下微丝结构趋于解聚。去除刺激后随时间延长 ,实验组逐渐恢复并比对照组增高 。 展开更多
关键词 微丝 形态结构 成骨细胞 细胞骨架 肌动蛋白 激光扫描共聚焦显微镜 机械压力
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2-BP对中性粒细胞趋化性的抑制作用 被引量:2
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作者 孙锦霞 王瑞 +2 位作者 谭笑 焦琛琛 黄钟 《深圳大学学报(理工版)》 EI CAS CSCD 北大核心 2017年第6期625-630,共6页
为研究棕榈酰化对中性粒细胞趋化性的调节作用,利用2-溴棕榈酸(2-bromopalmitate,2-BP)或棕榈酸(palmitate)预处理来源于骨髓的中性粒细胞.采用流式细胞术(flow cytometry,FCM)检测中性粒细胞的存活率和中性粒细胞内F-肌动蛋白(F-actin... 为研究棕榈酰化对中性粒细胞趋化性的调节作用,利用2-溴棕榈酸(2-bromopalmitate,2-BP)或棕榈酸(palmitate)预处理来源于骨髓的中性粒细胞.采用流式细胞术(flow cytometry,FCM)检测中性粒细胞的存活率和中性粒细胞内F-肌动蛋白(F-actin)的生成;通过免疫荧光分析中性粒细胞内F-actin的极化;利用细胞迁移试验检测中性粒细胞的迁移能力;通过免疫印迹的实验方法检测蛋白激酶B(protein kinase B,AKT)和磷酸化AKT的表达.结果发现,2-BP预处理显著抑制了中性粒细胞内F-actin极化,且抑制了中性粒细胞向趋化肽N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸(N-formylmethionyl-leucyl-phenylalanine,f MLP)迁移的能力,而棕榈酸预处理显著促进了中性粒细胞内F-肌动蛋白极化.两者对F-actin生成和AKT的磷酸化均无影响.研究结果表明,2-BP处理可能通过影响F-actin的极化抑制了中性粒细胞向f MLP的趋化活性. 展开更多
关键词 中性粒细胞 2-溴棕榈酸 F-肌动蛋白极化 趋化性 免疫荧光 流式细胞术 免疫印迹 细胞迁移 磷脂酰肌醇3-激酶-蛋白激酶B信号通路
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