DAO 66:33-40 (2005)  -  doi:10.3354/dao066033

Ultrastructural localization of antigenic determinants conserved during Perkinsus atlanticus trophozoite to prezoosporangium differentiation

Juan F. Montes*, Mercè Durfort, José García-Valero

Departament de Biologia Cel·lular, Facultat de Biologia, Universitat de Barcelona, Avda. Diagonal, 645, 08028 Barcelona, Spain
Centre de Referència de Recerca i Desenvolupament en Aqüicultura (CRAq) de la Generalitat de Catalunya

ABSTRACT: Trophozoite, prezoosporangium and zoospore are the 3 main developmental stages that form the life cycle of protozoa of the genus Perkinsus. Several studies have shown that the differentiation of Perkinsus species from the trophozoite to the prezoosporangium stage involves a substantial modification of the antigenic characteristics of these molluscan parasites. With the aim of determining the presence and distribution of antigenic determinants conserved during trophozoite to prezoosporangium differentiation, a polyclonal serum was raised against trophozoites of P. atlanticus purified from parasitized gills of the clam Tapes semidecussatus. Immunocytochemical analyses showed that the serum generated against P. atlanticus trophozoites strongly cross-reacted with the prezoosporangium stage. Immunogold electron microscopy studies revealed that the granular component of the nucleolus, chromatin, cell wall, plasmalemma, lomasomes and vacuolar membrane are the main subcellular structures where the immunodominant epitopes consistently expressed by trophozoites and prezoosporangia are located. Furthermore, analysis of the immunogold staining pattern revealed that the labelling density obtained for prezoosporangia in the nucleolus, cell wall, plasmalemma and lomasomes was significantly higher than that obtained for trophozoites. The most immunoreactive structure in trophozoites was the granular component of the nucleolus, whereas in prezoosporangia it was the lomasome. Interestingly, the main antigenic compartment of P. atlanticus, considering both developmental stages, was the lomasome of the prezoosporangium. These findings show that P. atlanticus trophozoite to prezoosporangium differentiation is accompanied by significant qualitative and quantitative changes in the ultrastructural distribution of the immunodominant antigens shared by these 2 developmental stages.

KEY WORDS: Perkinsus atlanticus · Protozoan parasite · Tapes semidecussatus · Clam disease · Antigenic cross-reactivity · Lomasomes · Electron microscopy · Immunocytochemistry

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