Localization of chitin in the cell wall of Candida albicans by means of wheat germ agglutinin. Fluorescence and ultrastructural studies.

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.