An assay to probe Plasmodium falciparum growth, transmission stage formation and early gametocyte development
An assay to probe Plasmodium falciparum growth, transmission stage formation and early gametocyte development
复制标题
DOI：
10.1038/nprot.2015.072
复制
发表时间：
2015-08-01
期刊：
影响因子：
14.8
通讯作者：
Marti, Matthias
中科院分区：
文献类型：
--
作者：
Brancucci, Nicolas M. B.;Goldowitz, Ilana;Marti, Matthias
从无性增殖到性分化的转变启动配子体的产生，这是人-蚊传播所需的疟原虫阶段。该方案描述了一种旨在探测药物或其他干扰对主要人类疟疾寄生虫恶性疟原虫的无性复制、性转换和早期配子体发育的影响的测定。通过添加条件培养基诱导同步无性复制的寄生虫产生配子体，然后在性承诺期间或在早期配子体发育的任何后续阶段将其暴露于感兴趣的处理。流式细胞术用于分别通过DNA染料染色和配子体特异性荧光蛋白表达来测量无性增殖和配子体产生。这种筛选方法可用于识别和评估潜在的传播阻断化合物，并进一步研究疟疾寄生虫的性转换机制。整个方案可在11 d内完成。
Conversion from asexual proliferation to sexual differentiation initiates the production of the gametocyte, which is the malaria parasite stage required for human-to-mosquito transmission. This protocol describes an assay designed to probe the effect of drugs or other perturbations on asexual replication, sexual conversion and early gametocyte development in the major human malaria parasite Plasmodium falciparum. Synchronized asexually replicating parasites are induced for gametocyte production by the addition of conditioned medium, and they are then exposed to the treatment of interest during sexual commitment or at any subsequent stage of early gametocyte development. Flow cytometry is used to measure asexual proliferation and gametocyte production via DNA dye staining and the gametocyte-specific expression of a fluorescent protein, respectively. This screening approach may be used to identify and evaluate potential transmission-blocking compounds and to further investigate the mechanism of sexual conversion in malaria parasites. The full protocol can be completed in 11 d.