RRC ID 1589
Author Sato Y, Shiraishi Y, Furuichi T.
Title Cell specificity and efficiency of the Semliki forest virus vector- and adenovirus vector-mediated gene expression in mouse cerebellum.
Journal J. Neurosci. Methods
Abstract 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.
Volume 137(1)
Pages 111-21
Published 2004-8-15
DOI 10.1016/j.jneumeth.2004.02.014
PII S0165027004000834
PMID 15196833
MeSH Adenoviridae / genetics* Adenoviridae / pathogenicity Animals Cell Line Cerebellar Cortex / cytology Cerebellar Cortex / metabolism Cerebellar Cortex / virology* Cricetinae Fetus Gene Expression Regulation, Viral / genetics Gene Transfer Techniques* Genetic Vectors / genetics* Green Fluorescent Proteins Luminescent Proteins / genetics Luminescent Proteins / metabolism Mice Mice, Inbred ICR Neural Pathways / cytology Neural Pathways / metabolism Neural Pathways / virology Neurons / cytology Neurons / metabolism Neurons / virology* Organ Culture Techniques / methods Purkinje Cells / cytology Purkinje Cells / metabolism Purkinje Cells / virology Reproducibility of Results Semliki forest virus / genetics* Semliki forest virus / pathogenicity Species Specificity Time Factors
IF 2.785
Times Cited 20
WOS Category BIOCHEMICAL RESEARCH METHODS NEUROSCIENCES
Resource
DNA material Ad5-dlX (RDB1747)