Cell specificity and efficiency of the Semliki forest virus vector- and adenovirus vector-mediated gene expression in mouse cerebellum.
Establishing efficient gene transfer and expression in post-mitotic neurons is important in understanding the genetic basis of neural circuits with cellular complexity. This study evaluates the properties of exogenous green fluorescent protein (GFP) expression mediated by the Semliki forest virus (SFV) and adenovirus (Ad) vectors in dissociated and slice cultures of the mouse cerebellum. Infection with SFV-GFP resulted in early-onset and high-level GFP expression in about 90% of Purkinje cells and in about 40% of granule cells in dissociated cultures at 1 day after infection. Two days after infection, GFP-positive cells showed signs of SFV-derived cytotoxicity. Ad-GFP infected almost all astrocytes and granule cells in dissociated cultures, and showed a steady increase in GFP fluorescence with a plateau at around 2 days post-infection. Ad vector-mediated GFP expression lasted for several weeks with no significant cell damage. In the slice cultures, both viral vectors mainly infected astroglial cells, but also showed a similar cell preference as that in dissociated cultures. These data indicate that the use of different viral vectors and infection conditions offers a powerful means of expressing exogenous genes in cerebellar cultures with different cell-type specificity and timing and duration of expression.
Sato
Yumi
Y
Laboratory for Molecular Neurogenesis, RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
Shiraishi
Yoko
Y
Furuichi
Teiichi
T
D000256:Adenoviridae / Q000235:genetics* / Q000472:pathogenicity; D000818:Animals; D002460:Cell Line; D002525:Cerebellar Cortex / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D006224:Cricetinae; D005333:Fetus; D015967:Gene Expression Regulation, Viral / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D009434:Neural Pathways / Q000166:cytology / Q000378:metabolism / Q000821:virology; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D009924:Organ Culture Techniques / Q000379:methods; D011689:Purkinje Cells / Q000166:cytology / Q000378:metabolism / Q000821:virology; D015203:Reproducibility of Results; D012672:Semliki forest virus / Q000235:genetics* / Q000472:pathogenicity; D013045:Species Specificity; D013997:Time Factors
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins