Localized gene transfer into organotypic hippocampal slice cultures and acute hippocampal slices

P Casaccia-Bonnefil, E Benedikz, H Shen, A Stelzer, D Edelstein, M Geschwind, M Brownlee, H J Federoff, P J Bergold

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Viral vectors derived from herpes simplex virus, type-1 (HSV), can transfer and express genes into fully differentiated, post-mitotic neurons. These vectors also transduce cells effectively in organotypic hippocampal slice cultures. Nanoliter quantities of a virus stock of HSVlac, an HSV vector that directs expression of E. coli beta-galactosidase (beta-gal), were microapplied into stratum pyramidale or stratum granulosum of slice cultures. Twenty-four hours later, a cluster of transduced cells expressing beta-gal was observed at the microapplication site. Gene transfer by microapplication was both effective and rapid. The titer of the HSVlac stocks was determined on NIH3T3 cells. Eighty-three percent of the beta-gal forming units successfully transduced beta-gal after microapplication to slice cultures. beta-Gal expression was detected as rapidly as 4 h after transduction into cultures of fibroblasts or hippocampal slices. The rapid expression of beta-gal by HSVlac allowed efficient transduction of acute hippocampal slices. Many genes have been transduced and expressed using HSV vectors; therefore, this microapplication method can be applied to many neurobiological questions.

TidsskriftJournal of Neuroscience Methods
Udgave nummer3
Sider (fra-til)341-51
Antal sider11
StatusUdgivet - dec. 1993
Udgivet eksterntJa