Abstract
To assess the dynamics of synthesis of the wall by regenerating Candida albicans protoplasts deposition of chitin and mannoproteins were investigated ultrastructurally using wheat germ agglutinin conjugated with either horseradish peroxidase or colloidal gold, and Concanavalin A coupled to ferritin respectively.
Freshly prepared protoplasts lacked wheat germ agglutinin receptor sites but after 1–2 h of regeneration, they were detected. After 4–5 h of regeneration, the cell wall showed a discrete structure which was only labelled with wheat germ agglutinin in thin sections. At this stage of regeneration the outermost layer of the wall was labelled with clusters of Concanavalin A-ferritin particles.
After 8 h regeneration, the cell wall appeared compact, and homogenously marked with wheat germ agglutinin whereas only the surface layers appeared consistently labelled with Concanavalin A-ferritin.
From these observations we conclude that C. albicans protoplasts are able to regenerate in liquid medium a cell wall consisting of a network of chitin fibrils and mannoproteins at least (glucan polymers were not determined in the present cytological study). The former are the fundamental component of the inner layers at early stages of regeneration, whereas the latter molecules are predominant in the outer layers of the wall.
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Abbreviations
- WGA-HRP:
-
wheat germ agglutinin conjugated with horseradish peroxidase
- WGA-Au:
-
wheat germ agglutinin conjugated with colloidal gold
- Con A-ferritin:
-
Concanavalin A coupled to ferritin
References
Boller, T, Durr M, Wiemken A (1976) Asymetric distribution of Concanavalin A binding sites on yeast plasmalemma and vacuolar membrane. Arch Microbiol 109:115–118
Duran A, Bowers B, Cabib E (1975) Chitin synthetase zymogen is attached to the yeast plasma membrane. Proc Natl Acad Sci USA 72:3952–3955
Elorza MV, Rico H, Sentandreu R (1983) Calcofuor white alters the assembly of chitin fibrils in Saccharomyces cerevisiae and Candida albicans cells. J Gen Microbiol 129:1577–1582
Elorza MV, Murgui A, Rico H, Miragall F, Sentandreu R (1987) Formation of a new cell wall by protoplasts of Candida albicans: Effect of papulacandin B, tunicamycin and nikkomicyn. J Gen Microbiol 133:2315–2325
Frens G (1973) Controlled nucleation for the regulation of particle size in monodisperse gold solutions. Nature 241:20–25
Gozalbo D, Dubón F, Sentandreu R (1985) Studies on zymogenity and solubilization of chitin synthase from Candida albicans. FEMS Microbiol Lett 26:59–63
Graham RC, Karnovsky JJ (1966) The early stages of absorption of injected horseradish peroxidase in proximal tubes of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291–302
Horisberger M, Volanthen M (1977) Localization of mannan and chitin on thin sections of budding yeast with gold markers. Arch Microbiol 115:1–7
Horisberger M, Rosset J, Bauer H (1976) Localization of mannan at the surface of yeast protoplasts by scanning electron microscopy. Arch Microbiol 109:9–14
Horisberger M, Rouvet-Vauthey M, Richli U, Farr D (1985) Cell wall architecture of the halophilic yeast Saccharomyces rouxii. An immunocytochemical study. Eur J Cell Biol 37:70–77
Lee KL, Buckley HR, Campbell C (1975) An aminoacid liquid synthetic medium for development of mycelial and yeast forms of Candida albicans. Sabouraudia 13:148–153
Miragall F, Rico H, Sentandreu R (1986) Changes in the plasma membrane of regenerating protoplasts of Candida albicans as revealed by freeze-fracture electron microscopy. J Gen Microbiol 132:2845–2853
Molano J, Bowers B, Cabib E (1980) Distribution of chitin in the yeast cell wall. An ultrastructural and chemical study. J Cell Biol 85:199–212
Murgui A, Elorza MW, Sentandreu R (1986) Tunicamycin and Paulacandin B inhibit incorporation of specific mannoproteins into the wall of Candida albicans regenerating protoplasts. Biochim Biophys Acta 884:550–558
Nečas O, Svoboda A (1985) Cell wall regeneration and protoplast reversion. In: Peberdy J, Ferenczy L (eds) Fungal protoplasts, vol 6. Marcel Dekker New York Basel, pp 115–133
Odds F (1985) Morphogenesis in Candida albicans. CRC Crit Rev Microbiol 12:45–93
Poulain D, Tronchin G, Dubremetz JF, Biguet J (1978) Ultrastructure of the cell wall of Candida albicans blastospores: Study of its constitutive layers by the use of a cytochemical technique revealing polysaccharides. Ann Microbiol (Institute Pasteur) 129A:141–153
Roberts RL, Bowers B, Slater ML, Cabib E (1983) Chitin synthesis and localization in cell division cycle mutants of Saccharomyces cerevisiae. Mol Cell Biol 3:922–930
Roth J, Bendayan M, Carlemaln M, Villinger W, Garavito M (1981) Enhancement of structural preservation and immuno-cytochemical staining in low temperature-embedded pancreatic tissue. J Histochem Cytochem 29:663–669
Santos E, Villanueva JR, Sentandreu R (1978) The plasma membrane of Saccharomyces cerevisiae: Isolation and some properties. Biochem Biophys Acta 508:39–45
Scarborough GA (1975) Isolation and characterization of Neurospora crassa plasma membranes. J Biol Chem 250:1106–1111
Sentandreu R, Herrero E, Elorza MV, Rico H, Pastor J (1983) Synthesis and assembly of wall polymers on regenerating yeast protoplasts. Experientia 46:187–195
Shepherd MG, Poulter RTM, Sullivan PA (1985) Candida albicans: biology, genetics, and pathogenicity. Ann Rev Microbiol 39:579–614
Spurr AR (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J Ultrastr Res 26:31–43
Tronchin G, Poulain D, Biguet J (1979) Etudes cytochimiques et ultrastructurales de la paroid de Candida albicans. I. Localization des mannanes par utilisation de Concanavalin A sur coupes ultrafines. Arch Microbiol 123:245–249
Tronchin G, Poulain D, Herbaut J, Biguet J (1981a) Cytochemical and ultrastructural studies of Candida albicans. II. Evidence for cell wall coat using Concanavalin A. J Ultrastr Res 75:50–59
Tronchin G, Poulain D, Herbaut J, Biguet J (1981b) Localization of chitin in the cell wall of Candida albicans by means of wheat germ agglutinin. Fluoresence and ultrastructural studies. Eur J Cell Biol 26:121–128
Tronchin G, Poulain D, Herbaut J, Biguet J (1982) Aspect ultrastructuraux et cytochimiques de la regeneration des spheroplastes chez Candida albicans. Ann Microbiol (Institute Pasteur) 133A:276–291
Van der Valk P, Wessels JGH (1976) Ultrastructure and localization of wall polymers during regeneration and reversion of protoplasts of Schizophyllum comune. Protoplasma 90:65–87
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Miragall, F., Rico, H. & Sentandreu, R. Regeneration of the cell wall in protoplasts of Candida albicans . Arch Microbiol 149, 286–290 (1988). https://doi.org/10.1007/BF00411643
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DOI: https://doi.org/10.1007/BF00411643