Modified herpes simplex virus delivery of enhanced GFP into the central nervous system.
Controlled expression of proteins is a key experimental approach to a deeper understanding of the molecular basis of neuronal function. Here we evaluate the HSV-1 (herpes simplex virus) amplicon vector for gene delivery into the brains of living rats. We demonstrate that HSV-1 amplicon vectors expressing enhanced green fluorescent protein (EGFP) can reliably infect neurons after it is injected into cortex, striatum and thalamus in rats, producing sufficient numbers of infected neurons for electrophysiological experiments in acute brain slices. Expression of EGFP delivered by the HSV-1 amplicon was detected for up to 5 weeks post-infection. We detected no changes in the morphology or the electrophysiological properties of thalamic, striatal or cortical neurons within a period of at least 2 weeks after HSV-1 amplicon injection. We conclude that the HSV-1 amplicon is a valuable tool for gene delivery in the rat central nervous system.
Sandler
Vladislav M
VM
HHMI, Children's Hospital, Boston, MA, USA
Wang
Samuel
S
Angelo
Kamilla
K
Lo
Helen G
HG
Breakefield
Xandra O
XO
Clapham
David E
DE
D000818:Animals; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D005784:Gene Amplification; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D018259:Herpesvirus 1, Human / Q000235:genetics*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008564:Membrane Potentials / Q000502:physiology; D018408:Patch-Clamp Techniques / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D014161:Transduction, Genetic
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins