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

Patrizia Casaccia-Bonnefil, Eirikur Benedikz, Hong Shen, Armin Stelzer, Diane Edelstein, Michael Geschwind, Michael Brownlee, Howard J. Federoff, Peter J. Bergold

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

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 β-galactosidase (β-gal), were microapplied into stratum pyramidale or stratum granulosum of slice cultures. Twenty-four hours later, a cluster of transduced cells expressing β-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 β-gal forming units successfully transduced β-gal after microapplication to slice cultures. β-Gal expression was detected as rapidly as 4 h after transduction into cultures of fibroblasts or hippocampal slices. The rapid expression of β-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.

Original languageEnglish (US)
Pages (from-to)341-351
Number of pages11
JournalJournal of Neuroscience Methods
Volume50
Issue number3
DOIs
StatePublished - Dec 1993

Keywords

  • Gene transduction
  • Genetic vector
  • Herpes simplex virus-1
  • Microinjection
  • beta-Galactosidase

ASJC Scopus subject areas

  • Neuroscience(all)

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    Casaccia-Bonnefil, P., Benedikz, E., Shen, H., Stelzer, A., Edelstein, D., Geschwind, M., Brownlee, M., Federoff, H. J., & Bergold, P. J. (1993). Localized gene transfer into organotypic hippocampal slice cultures and acute hippocampal slices. Journal of Neuroscience Methods, 50(3), 341-351. https://doi.org/10.1016/0165-0270(93)90040-X