Confocal Laser Scanning Microscopy of β-Tubulin and α-Actinin in Asexual Stages of Eimeria tenella (Apicomplexa: Eimeriidae) in Cell Culture

Abstract

Summary: Specific monoclonal antibodies were used to visualize the dynamics of β-tubulin and α-actinin in asexual development of Eimeria tenella in cultured primary chicken kidney cells by means of confocal laser scanning microscopy. β-tubulin was more condensed in refractile bodies (RB), at anterior ends, and along subpellicles in sporozoites. The RB remained labelled during the development of first generation schizonts until the formation of merozoites, suggesting a possible important role of RB in the reorganization of microtubules in schizonts. The microtubules also formed a number of tiny aggregation centers scattered throughout the schizonts corresponding to the formation of first generation merozoites. The microtubular material in second generation schizonts was clustered around several weakly-stained areas. Both first and second generation merozoites had more highly concentrated β-tubulin at anterior ends and along subpellicles. In contrast, α-actinin, and actin-binding protein, was more concentrated in the anterior half area of sporozoites. It became evenly distribution in the whole cytosol of developing trophozoites except RB areas. The distribution of α-actinin in first or second generation schizonts corresponded to merozoite formation. The β-tubulin and α-actinin of chicken primary kidney cells were also labelled by the two monoclonal antibodies.