Yokoyama K, Kaji H, Nishimura K, Miyaji M
Department of Fungal Infections, Chiba University, Japan.
J Gen Microbiol. 1990 Jun;136(6):1067-75. doi: 10.1099/00221287-136-6-1067.
Cytoskeleton inhibitors were used to study morphogenesis in the pathogenic and dimorphic fungus Candida albicans. Nocodazole is a specific microtubule inhibitor and chloropropham (CIPC), at high concentrations, is an inhibitor of microtubules and microfilaments. Distribution of microtubules and microfilaments was studied by immunofluorescence techniques using anti-tubulin antibody with FITC-conjugated secondary antibody, and by staining with Rh-phalloidin. Nocodazole did not arrest apical cell elongation at a concentration (20 micrograms ml-1) that inhibited nuclear division and migration. Cytoplasmic and nuclear microtubules disappeared within 30 min in filamentous cells under these conditions. However, the Rh-phalloidin-stained actin granules which were localized in the tips of filamentous cells, and the microfilaments, were arranged normally at this concentration of nocodazole. Growth, and normal distribution of microtubules and microfilaments, were inhibited by a high concentration (200 micrograms ml-1) of CIPC. At a concentration (100 micrograms ml-1) of CIPC that permitted nuclear division, apical cell elongation was arrested, and filamentous growth was converted into yeast growth. At this concentration of CIPC, microtubules were distributed normally in filamentous cells. Long microfilaments were not observed, and actin granules did not localize in the tips of filamentous cells, but were distributed throughout the cytoplasmic cortex. Our results show that cytoplasmic microtubules are not essential for the elongation of filamentous cell tips but that microfilaments are apparently essential for this process.
细胞骨架抑制剂被用于研究致病性双态真菌白色念珠菌的形态发生。诺考达唑是一种特异性微管抑制剂，而氯苯胺灵（CIPC）在高浓度时是微管和微丝的抑制剂。使用抗微管蛋白抗体与异硫氰酸荧光素（FITC）偶联的二抗，通过免疫荧光技术，以及用罗丹明鬼笔环肽染色，来研究微管和微丝的分布。在抑制核分裂和迁移的浓度（20微克/毫升）下，诺考达唑并未阻止顶端细胞伸长。在这些条件下，丝状细胞中的细胞质和核微管在30分钟内消失。然而，在这个诺考达唑浓度下，位于丝状细胞顶端的罗丹明鬼笔环肽染色的肌动蛋白颗粒以及微丝排列正常。高浓度（200微克/毫升）的CIPC抑制生长以及微管和微丝的正常分布。在允许核分裂的CIPC浓度（100微克/毫升）下，顶端细胞伸长被阻止，丝状生长转变为酵母生长。在这个CIPC浓度下，微管在丝状细胞中正常分布。未观察到长微丝，肌动蛋白颗粒不在丝状细胞顶端定位，而是分布在整个细胞质皮层。我们的结果表明，细胞质微管对于丝状细胞顶端的伸长不是必需的，但微丝显然对于这个过程是必需的。
