Gene transfer into neurons from hippocampal slices: comparison of recombinant Semliki Forest Virus, adenovirus, adeno-associated virus, lentivirus, and measles virus.
Viral vectors are useful for transferring genes into neurons. Here, we characterized recombinant Semliki Forest virus (SFV), adenovirus type 5 (Ad5), adeno-associated virus type 2 (AAV), lentivirus, and measles virus (MV) by their expression of green fluorescent protein (GFP) in rat hippocampal slice cultures. SFV infected more neurons (>90% of all GFP-positive cells) than AAV, lentivirus, and MV (71, 69, and 62%, respectively), whereas no infected neurons were identified with Ad5. AAV-mediated GFP expression was neuron-specific when the platelet-derived growth factor beta-chain promoter rather than cytomegalovirus promoter was used. Transgene expression occurred rapidly but transiently for SFV, increased slowly but remained stable with AAV and lentivirus, and was fast with MV. Resting membrane potential and conductance, action potentials, firing accommodation, and H-current appeared normal in infected CA1 pyramidal cells. Thus, SFV is useful for short-term and AAV and lentivirus for long-term transduction of hippocampal slices, while MV constitutes a novel vector.
Ehrengruber
M U
MU
Brain Research Institute, Institute of Molecular Biology, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. ehrengru@hifo.unizh.ch
Hennou
S
S
Büeler
H
H
Naim
H Y
HY
Déglon
N
N
Lundstrom
K
K
D000256:Adenoviridae / Q000235:genetics / Q000472:pathogenicity; D000818:Animals; D002470:Cell Survival / Q000235:genetics; D000229:Dependovirus / Q000235:genetics / Q000472:pathogenicity; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000502:physiology*; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism / Q000821:virology; D007202:Indicators and Reagents / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics / Q000472:pathogenicity; D008164:Luminescent Proteins / Q000378:metabolism; D008459:Measles virus / Q000235:genetics / Q000472:pathogenicity; D008564:Membrane Potentials / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D009924:Organ Culture Techniques; D017966:Pyramidal Cells / Q000166:cytology / Q000502:physiology / Q000821:virology; D051381:Rats; D012672:Semliki forest virus / Q000235:genetics / Q000472:pathogenicity; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes / Q000502:physiology*; D014774:Virulence / Q000235:genetics; D014780:Viruses / Q000235:genetics* / Q000472:pathogenicity
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins