Abstract:

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.

共聚焦荧光镜检术揭示大蒜鳞片细胞间期核中的F肌动蛋白网络
王冬梅1,2  王学臣2* 张伟成3

（1.  河北农业大学农学院，保定071001; 2.  中国农业大学生物学院，北京100094
3.  中国科学院上海植物生理研究所，上海200032）

摘要：采用免疫荧光和荧光分子探针技术与共聚焦激光扫描显微镜观察相结合，对大蒜（Ａｌｌｉｕｍ ｓａｔｉｖｕｍ Ｌ．）鳞片细胞间期核中是否存在Ｆ肌动蛋白进行了研究。结果表明，以兔抗肌动蛋折抗体为一抗、ＦＴＴＧ－羊抗兔ＩｇＧ抗体为二抗进行免疫荧光标记实验，在荧光镜下观察到蒜瓣薄壁组织的细胞核及表皮细胞核均发出明亮的黄绿色荧光经共聚焦激光扫描显微镜进一步检查，整个细胞核呈黄绿色荧光，说明其中含有肌动蛋白。经ＴＲITC-鬼笔环肽（TRITC-phalloidin）标记后，细胞核散发出明亮的红色荧光，经共聚焦激光扫描显微镜做光切片及三维重组图像，发现核中有一个三维网络结构。细胞松弛素D（cytochalasin D）预处理后，核中三维网络结构不复存在，但核仍显示红色荧光；若经非标phalloidin预处理，则核中几乎看不到红色荧光和任何网络残迹。对照实验结果进一步表明，*肌动蛋白存在于大蒜细胞核中并以* 肌动蛋白纤束形成三维网络结构。
关键词： 间期核；F 肌动蛋白；异硫氰四甲基罗丹明-鬼笔环肽；细胞松弛素D；共聚焦激光扫描显微镜；大蒜

Key words: interphase nucleus, F-actin, TRITC-phalloidin, cytochalasin D, confocal laser scanning microscopy, Allium sativum