Telomeres Laboratory, Department of Chemical and Biological Sciences, Biosciences Institute, São Paulo State University (UNESP), Botucatu, SP, Brazil.
DNA Replication and Repair Laboratory (DRRL), Department of Chemical and Biological Sciences, Biosciences Institute, São Paulo State University (UNESP), Botucatu, SP, Brazil.
Methods Mol Biol. 2022;2579:127-135. doi: 10.1007/978-1-0716-2736-5_10.
Leishmania spp. comprises a group of protozoan parasites that affect millions of people around the world. Understanding the main cell cycle-dependent events could provide an important route for developing specific therapies since some factors involved in cell cycle control may have low similarity relative to their homologs in mammals. Furthermore, accurate cell cycle-dependent analyses often require many cells, which can be achieved through cell cycle synchronization. Here, we described a useful method to synchronize procyclic promastigote forms of Leishmania amazonensis using hydroxyurea (HU) and the analysis of its DNA content profile. This approach can be extended to other trypanosomatids, such as Trypanosoma cruzi or Trypanosoma brucei, and provides an effective method for arresting more than 80% of cells at the G1/S phase transition.
利什曼原虫属包含一组原生动物寄生虫，影响着全球数百万人。了解主要的细胞周期依赖性事件可能为开发特定疗法提供重要途径，因为细胞周期控制中涉及的一些因素与哺乳动物中的同源物相比可能具有较低的相似性。此外，准确的细胞周期依赖性分析通常需要许多细胞，这可以通过细胞周期同步来实现。在这里，我们描述了一种使用羟基脲 (HU) 同步美洲利什曼原虫前鞭毛体形式的有用方法，并分析了其 DNA 含量图谱。这种方法可以扩展到其他原生动物，如克氏锥虫或布氏锥虫，并为在 G1/S 期转换时阻止超过 80%的细胞提供了一种有效方法。
