Tronchin G, Poulain D, Herbaut J, Biguet J
Eur J Cell Biol. 1981 Dec;26(1):121-8.
Cytochemical localization of wheat germ agglutinin binding sites in the cell wall of Candida albicans was investigated with fluorescence and electron microscopy. Various analytical techniques were employed in order to obtain a good penetration of the cytochemical markers, glycosylated horseradish peroxidase or glycosylated ferritin. In blastospores sectioned by cryostatic methods, a weak and continuous labelling of the blastospore periphery was observed with peroxidase, whereas bud scars and inner cell wall areas were labelled with ferritin. Following enzymatic treatment with pronase whose efficiency was followed by the periodic acid-thiocarbohydrazide -- Silver proteinate technique, the inner cell wall layers of bud are strongly stained with both fluorescein and the reactions products of peroxidase. After pronase-chitinase treatment, fluorescence was observed only in the mother cell wall. Finally, ultrathin glycol methacrylate sections showed a labelling both in inner and outer layers. All these results suggest that chitin was essentially distributed in the inner wall layers near the plasmalemma and in a smaller amount in outer wall layers. On the basis of the present findings, a hypothesis of wall assembly is proposed.
利用荧光显微镜和电子显微镜研究了小麦胚凝集素在白色念珠菌细胞壁上结合位点的细胞化学定位。采用了各种分析技术，以便使细胞化学标记物（糖基化辣根过氧化物酶或糖基化铁蛋白）能良好地渗透。在用低温切片法制备的芽生孢子切片中，过氧化物酶可观察到芽生孢子周边有微弱且连续的标记，而芽痕和细胞壁内层则被铁蛋白标记。在用链霉蛋白酶进行酶处理后（其效率通过高碘酸 - 硫代碳酰肼 - 蛋白银技术进行跟踪），芽的细胞壁内层被荧光素和过氧化物酶的反应产物强烈染色。经过链霉蛋白酶 - 几丁质酶处理后，仅在母细胞壁中观察到荧光。最后，超薄甲基丙烯酸乙二醇酯切片显示在内层和外层均有标记。所有这些结果表明，几丁质主要分布在靠近质膜的内壁层，在外壁层中的含量较少。基于目前的研究结果，提出了细胞壁组装的假说。
