Chen Jie, Guo Wei, Dang Xiaoqun, Huang Yukang, Liu Fangyan, Meng Xianzhi, An Yaoyao, Long Mengxian, Bao Jialing, Zhou Zeyang, Xiang Zhonghuai, Pan Guoqing
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, P. R. China.
Key Laboratory of Sericultural Biology and Genetic Breeding, Ministry of Agriculture, Chongqing, P. R. China.
PLoS One. 2017 Jun 22;12(6):e0179618. doi: 10.1371/journal.pone.0179618. eCollection 2017.
Microsporidia are eukaryotic, unicellular parasites that have been studied for more than 150 years. These organisms are extraordinary in their ability to invade a wide range of hosts including vertebrates and invertebrates, such as human and commercially important animals. A lack of appropriate labeling methods has limited the research of the cell cycle and protein locations in intracellular stages. In this report, an easy fluorescent labeling method has been developed to mark the proliferative and sporogonic phases of microsporidia Nosema bombycis in host cells. Based on the presence of chitin, Calcofluor White M2R was used to label the sporogonic phase, while β-tubulin antibody coupled with fluorescence secondary antibody were used to label the proliferative phase by immunofluorescence. This method is simple, efficient and can be used on both infected cells and tissue slices, providing a great potential application in microsporidia research.
微孢子虫是真核单细胞寄生虫，对其研究已超过150年。这些生物具有非凡的能力，能够侵入包括脊椎动物和无脊椎动物（如人类和具有重要商业价值的动物）在内的广泛宿主。缺乏合适的标记方法限制了对细胞周期和细胞内阶段蛋白质定位的研究。在本报告中，已开发出一种简便的荧光标记方法，用于标记宿主细胞中家蚕微孢子虫的增殖期和孢子生殖期。基于几丁质的存在，使用荧光增白剂M2R标记孢子生殖期，而β-微管蛋白抗体与荧光二抗结合，通过免疫荧光标记增殖期。该方法简单、高效，可用于感染细胞和组织切片，在微孢子虫研究中具有巨大的潜在应用价值。
