David Marek, Gabriel Miroslav, Kopecká Marie
Department of Biology, Faculty of Medicine, Masaryk University, Tomesova 12, Brno 602 00, Czech Republic.
Cell Biol Int. 2007 Jan;31(1):16-23. doi: 10.1016/j.cellbi.2006.09.001. Epub 2006 Sep 10.
Microtubular and actin cytoskeletons were investigated in the lipophilic yeast Malassezia pachydermatis by fluorescence and electron microscopy. To detect microtubules by indirect immunofluorescence using monoclonal anti-tubulin antibody, a prolonged incubation with lysing enzymes was necessary due to its very thick cell wall. Cytoplasmic microtubules were detected in interphase and a spindle with astral microtubules was seen in M-phase. The disintegration of cytoplasmic microtubules and migration of the nucleus to the bud before mitosis were characteristic features of the basidiomycetous yeast Malassezia pachydermatis. The visualisation of F-actin structures (patches, cables and cytokinetic rings) by fluorescence microscopy using both monoclonal anti-actin antibody and rhodamine-phalloidin failed, but actin was detected by electron microscopy with immunogold labelling. Clusters of gold particles indicating actin structures were detected at the plasma membrane of cells with unique cortical ultrastructural features characteristic of the genus Malassezia. A possible association of these with the actin cytoskeleton is suggested.
通过荧光显微镜和电子显微镜研究了亲脂性酵母厚皮马拉色菌中的微管和肌动蛋白细胞骨架。由于厚皮马拉色菌细胞壁非常厚,因此使用单克隆抗微管蛋白抗体通过间接免疫荧光检测微管时,需要用裂解酶长时间孵育。在间期检测到细胞质微管,在M期观察到带有星状微管的纺锤体。细胞质微管的解体以及有丝分裂前细胞核向芽的迁移是担子菌酵母厚皮马拉色菌的特征性特征。使用单克隆抗肌动蛋白抗体和罗丹明-鬼笔环肽通过荧光显微镜观察F-肌动蛋白结构(斑块、束和细胞分裂环)未成功,但通过免疫金标记的电子显微镜检测到了肌动蛋白。在具有马拉色菌属独特皮质超微结构特征的细胞的质膜上检测到指示肌动蛋白结构的金颗粒簇。提示这些可能与肌动蛋白细胞骨架有关。