TY - JOUR
T1 - SYTO11 staining vs FISH staining
T2 - A comparison of two methods to stain Wolbachia pipientis in cell cultures
AU - Venard, C. M.P.
AU - Crain, P. R.
AU - Dobson, S. L.
PY - 2011/2
Y1 - 2011/2
N2 - Aims: The Aedes albopictus C7-10 cell line was infected with Wolbachia strains wRi and wAlbB to create C7-10R and C7-10B cell lines, respectively. We compared two different methods, fluorescence in situ hybridization staining and SYTO11 staining, to describe these new Wolbachia infections in C7-10.Methods and Results: Both staining methods were as efficient to stain Wolbachia. A formula was developed to quantify Wolbachia infection. The infection levels in C7-10B and C7-10R differed. The live stain SYTO11 was found to be useful to visualize Wolbachia in replicating host cells. Its potential cytotoxic effect at high concentration was investigated.Conclusions: C7-10 supported two Wolbachia infections, constituting new tools to study Wolbachia-host interactions. The different infection levels suggest that wRi and wAlbB have different requirements for their survival in C7-10 host cell line. Observation of SYTO11-stained live cells gave new insights on Wolbachia segregation 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 of Wolbachia's intracellular lifestyle. The Wolbachia infections in C7-10 described here could be used as in vitro models to investigate Wolbachia biology.
AB - Aims: The Aedes albopictus C7-10 cell line was infected with Wolbachia strains wRi and wAlbB to create C7-10R and C7-10B cell lines, respectively. We compared two different methods, fluorescence in situ hybridization staining and SYTO11 staining, to describe these new Wolbachia infections in C7-10.Methods and Results: Both staining methods were as efficient to stain Wolbachia. A formula was developed to quantify Wolbachia infection. The infection levels in C7-10B and C7-10R differed. The live stain SYTO11 was found to be useful to visualize Wolbachia in replicating host cells. Its potential cytotoxic effect at high concentration was investigated.Conclusions: C7-10 supported two Wolbachia infections, constituting new tools to study Wolbachia-host interactions. The different infection levels suggest that wRi and wAlbB have different requirements for their survival in C7-10 host cell line. Observation of SYTO11-stained live cells gave new insights on Wolbachia segregation 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 of Wolbachia's intracellular lifestyle. The Wolbachia infections in C7-10 described here could be used as in vitro models to investigate Wolbachia biology.
KW - FISH staining
KW - Insect cell culture
KW - SYTO11 staining
KW - Wolbachia pipentis
UR - http://www.scopus.com/inward/record.url?scp=78650973227&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=78650973227&partnerID=8YFLogxK
U2 - 10.1111/j.1472-765X.2010.02986.x
DO - 10.1111/j.1472-765X.2010.02986.x
M3 - Article
C2 - 21214605
AN - SCOPUS:78650973227
SN - 0266-8254
VL - 52
SP - 168
EP - 176
JO - Letters in Applied Microbiology
JF - Letters in Applied Microbiology
IS - 2
ER -