Rapid and stable gene expression in hippocampal slice cultures from a defective HSV-1 vector.
Stable transfer of genetic information into neurons is a powerful strategy to elucidate specific mechanisms of neurophysiology and to develop therapies for neurological disorders. To evaluate the optimal parameters for efficient gene delivery of defective herpes simplex virus type one (HSV-1) vectors into a specific brain region, an HSV-1 vector expressing E. coli beta-galactosidase was used to infect organotypic cultures of hippocampal slices. beta-Galactosidase was expressed as early as 2 h after infection in a dose-dependent manner as measured on immunoblots, and reached a maximum level after approximately 35 h. Expression of the RNA and the antigen was still evident after the longest time sampled (11-12 days), whereas no beta-galactosidase was ever detected in cultured slices infected with a control virus lacking the reporter gene. Hippocampal cells expressing the reporter gene outlined the contour of the neuronal cell body layers in fields CA3 and dentate gyrus; such correspondence was less evident in field CA1. Anatomical, morphological, and immunohistochemical criteria also confirmed that the majority of these infected cells were neurons. beta-Galactosidase was also detected in the somata and processes of infected interneurons. Tests for synaptic pathology associated with virus infection showed no changes in pre- and postsynaptic markers.
Bahr
B A
BA
Center for the Neurobiology of Learning and Memory, University of California, Irvine 92717.
Neve
R L
RL
Sharp
J
J
Geller
A I
AI
Lynch
G
G
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D003673:Defective Viruses; D004926:Escherichia coli; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human; D006624:Hippocampus / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009924:Organ Culture Techniques; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D013997:Time Factors; D001616:beta-Galactosidase / Q000096:biosynthesis*
D017931:DNA Primers; D012329:RNA, Bacterial; D001616:beta-Galactosidase