1.Department of Pathology， The Central Hospital of Wuhan， Tongji Medical College， Huazhong University of Science and Technology Wuhan 430014， China；2.Brain and Cognition Research Institute， Wuhan University of Science and Technology， Wuhan 430065， China；3.Center for Brain Science， Wuhan Institute of Physics and Mathematics， Chinese Academy of Sciences， Wuhan 430071， China
Abstract：Pseudorabies virus (PRV) is one of the commonly used neurotropical viruses for neural circuits tracing studies. However， the affects of preservation conditions of PRV on its infection titer and in vivo infection efficiency remains unclear. In this study， 2 μL aliquots of PRV with 1-4 times of freeze-thaw cycles at －80℃ or room temperature within 2 h were respectively tittered on BHK21 cells using plaque methods. The results showed that 1-3 times of freeze-thaw cycles had no significant effect on the PRV titers， while the fourth freeze-thaw cycles significantly reduced the PRV titers. Then， 2 μL PRV aliquots were subjected to one freezing-thawing cycle and stored at －80℃ and －20℃， respectively. Two weeks later， they were separately tittered on BHK21 cells and then stereotacticly injected into the dentate gyrus (DG) area of mice for in vivo neural circuit tracing analyzation. We found that the viral titer and in vivo tracing efficiency of PRV were significantly affected by the long-time storage temperatures. Our results may provide some helpful instructions for the preservation of PRV virus and its applications in neural circuit mapping.
2019, Vol. 53 
