Developmental stages identified in the trophozoite of the free-living Alveolate flagellate < i > Colpodella < /i > sp. (Apicomplexa)

In this study we performed light, immunofluorescent and transmission electron microscopy ofColpodella trophozoites to characterize trophozoite morphology and protein distribution. The use of Giemsa staining and antibodies to distinguishColpodella life cycle stages has not been performed previously. Rhoptry andβ-tubulin antibodies were used in immunofluorescent assays (IFA) to identify protein localization and distribution in the trophozoite stage ofColpodella (ATCC 50594). We report novel data identifying“doughnut-shaped” vesicles in the cytoplasm and apical end ofColpodella trophozoites reactive with antibodies specific toPlasmodium merozoite rhoptry proteins. Giemsa staining and immunofluorescent microscopy identified different developmental stages ofColpodella trophozoites, with the presence or absence of vesicles corresponding to maturity of the trophozoite. These data demonstrate for the first time evidence of rhoptry protein conservation betweenPlasmodium andColpodella and provide further evidence thatColpodella trophozoites can be used as a heterologous model to investigate rhoptry biogenesis and function. Staining and antibody reactivity will facilitate phylogenetic, biochemical and molecular investigations ofColpodella sp. Developmental stages can be distinguished by Giemsa staining and antibody reactivity.Keywords: Colpodella· Rhoptries· Trichocysts· Apical complex· Plasmodium RhopH3

Source: International Microbiology - March 26, 2018 Category: Microbiology Source Type: research