Identification Of Recombinant Adenovirus Carrying Mouse Estrogen Receptor α In Infected Neurons And Central Nervous System

Objective:Recombinant adenovirus carrying C57BL/6 mouse estrogen receptor a (Era) was used to infect mouse neurons. The purpose of this study is to lay the foundation for further study of the role of Era in the development of central nervous system and the relationship between Era and some nervous system diseases.Methods:The nerve cells of C57BL/6 mouse were cultured using a serum-free culture medium. The purity of neurons was confirmed by immunohistochemistry. Recombinant adenovirus carrying C57BL/6 mouse Era was used to infect mouse nerve cells, and the infection efficiency and apoptosis rate were examined by flow cytometry(FCM)to obtain optimal multiplicity of infection (MOI). The expression of ErαmRNA and protein in infected nerve cells were detected by quantitative real-time PCR and western blot. Application of three-dimensional localization in vivo infection of the central nervous system, observe the fluorescence expression.Results:Nerve cells was successfully primary cultured and the purity was over 90%. MOI=100 can be used as optimal MOI in nerve cells infected by Erarecombinant adenovirus, and the infection efficiency amounted to 84.42±4.47% but cell apoptosis rate was only 3.62±0.72%. The recombinant adenovirus carring Era infected nerve cells successfully and the expression of Era mRNA and protein were higher in infected nerve cells.Conclusions:The recombinant adenovirus carring Era infected nerve cells and central nervous system successfully leading to Era mRNA and protein significant upregulation.