SYTO11 staining vs FISH staining: a comparison of two methods to stain Wolbachia pipientis in cell cultures.
SYTO11 staining vs FISH staining: a comparison of two methods to stain Wolbachia pipientis in cell cultures.
复制标题
SYTO11 染色与 FISH 染色：细胞培养物中两种 Wolbachia pipientis 染色方法的比较。
DOI：
10.1111/j.1472-765x.2010.02986.x
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发表时间：
2011
影响因子：
2.4
通讯作者：
Dobson,SL
中科院分区：
文献类型：
--
作者：
Venard,CM-P;Crain,PR;Dobson,SL
目的：用Wolbachirin inswRi和wAlbB分别感染白纹伊蚊C7 - 10细胞系，建立C7 - 10R和C7 - 10B细胞系。我们比较了两种不同的方法，荧光原位杂交染色和SYTO 11染色，以描述这些新的Wolbachia感染在C7 - 10。方法和结果：两种染色方法是有效的染色Wolbachia。建立了定量沃尔巴克氏体感染的公式。C7 - 10B和C7 - 10R的感染水平不同。发现活染色剂SYTO 11可用于观察沃尔巴克氏体复制宿主细胞。结论：C7 - 10支持两种Wolbachia感染，为研究Wolbachia-宿主相互作用提供了新的工具。不同的感染水平表明wRi和wAlbB对它们在C7 - 10宿主细胞系中的存活有不同的要求。SYTO 11染色活细胞的观察为宿主细胞有丝分裂过程中的沃尔巴克氏体分离模式提供了新的见解。研究的意义和影响：沃尔巴克氏体诱导的节肢动物和蠕虫宿主表型可能用于控制害虫种群。然而，由于沃尔巴克氏体的细胞内生活方式，这些表型的机制很难研究。本文描述的C7 - 10中的Wolbachiaininfection可用作研究Wolbachiabiology的体外模型。
Aims:TheAedes albopictusC7‐10 cell line was infected withWolbachiastrainswRi andwAlbB to create C7‐10R and C7‐10B cell lines, respectively. We compared two different methods, fluorescencein situhybridization staining and SYTO11 staining, to describe these newWolbachiainfections in C7‐10.Methods and Results:Both staining methods were as efficient to stainWolbachia. A formula was developed to quantifyWolbachiainfection. The infection levels in C7‐10B and C7‐10R differed. The live stain SYTO11 was found to be useful to visualizeWolbachiain replicating host cells. Its potential cytotoxic effect at high concentration was investigated.Conclusions:C7‐10 supported twoWolbachiainfections, constituting new tools to studyWolbachia–host interactions. The different infection levels suggest thatwRi andwAlbB have different requirements for their survival in C7‐10 host cell line. Observation of SYTO11‐stained live cells gave new insights onWolbachiasegregation pattern during host cell mitosis.Significance and Impact of the Study:Wolbachia‐induced phenotypes in their arthropod and worm hosts could potentially be used to control pest populations. However, the mechanisms underlying these phenotypes are difficult to study because ofWolbachia’s intracellular lifestyle. TheWolbachiainfections in C7‐10 described here could be used asin vitromodels to investigateWolbachiabiology.