Department of Biological, Geological and Environmental Sciences, Cleveland State University, 2121 Euclid Avenue, SI 219 Cleveland, Ohio 44115.
Int Microbiol. 2017 Dec;20(4):178-183. doi: 10.2436/20.1501.01.301.
In this study we performed light, immunofluorescent and transmission electron microscopy of Colpodella trophozoites to characterize trophozoite morphology and protein distribution. The use of Giemsa staining and antibodies to distinguish Colpodella life cycle stages has not been performed previously. Rhoptry and β-tubulin antibodies were used in immunofluorescent assays (IFA) to identify protein localization and distribution in the trophozoite stage of Colpodella (ATCC 50594). We report novel data identifying "doughnut-shaped" vesicles in the cytoplasm and apical end of Colpodella trophozoites reactive with antibodies specific to Plasmodium merozoite rhoptry proteins. Giemsa staining and immunofluorescent microscopy identified different developmental stages of Colpodella 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 between Plasmodium and Colpodella and provide further evidence that Colpodella 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 of Colpodella sp. Developmental stages can be distinguished by Giemsa staining and antibody reactivity.
在这项研究中，我们对 Colpodella 滋养体进行了光镜、免疫荧光和透射电镜检查，以描述滋养体的形态和蛋白分布。以前没有使用吉姆萨染色和抗体来区分 Colpodella 的生命周期阶段。在免疫荧光分析（IFA）中使用了 泡状结构蛋白和β-微管蛋白抗体来识别 Colpodella 滋养体阶段的蛋白定位和分布（ATCC 50594）。我们报告了新的数据，这些数据表明在 Colpodella 滋养体的细胞质和顶端反应与疟原虫裂殖子泡状结构蛋白特异性抗体的“甜甜圈状”囊泡。吉姆萨染色和免疫荧光显微镜鉴定了 Colpodella 滋养体的不同发育阶段，囊泡的存在或缺失与滋养体的成熟程度相对应。这些数据首次证明了疟原虫和 Colpodella 之间泡状结构蛋白的保守性，并进一步证明 Colpodella 滋养体可用作研究泡状结构生物发生和功能的异源模型。染色和抗体反应将有助于 Colpodella sp 的系统发育、生化和分子研究。发育阶段可以通过吉姆萨染色和抗体反应来区分。
