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 Citations (Scopus)

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 - 1993

Fingerprint

Human Herpesvirus 1
Galactosidases
Genes
Fibroblasts
Escherichia coli
Viruses
Neurons

Keywords

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

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Casaccia-Bonnefil, P., Benedikz, E., Shen, H., Stelzer, A., Edelstein, D., Geschwind, M., ... 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

Localized gene transfer into organotypic hippocampal slice cultures and acute hippocampal slices. / Casaccia-Bonnefil, Patrizia; Benedikz, Eirikur; Shen, Hong; Stelzer, Armin; Edelstein, Diane; Geschwind, Michael; Brownlee, Michael; Federoff, Howard J.; Bergold, Peter J.

In: Journal of Neuroscience Methods, Vol. 50, No. 3, 1993, p. 341-351.

Research output: Contribution to journalArticle

Casaccia-Bonnefil, P, Benedikz, E, Shen, H, Stelzer, A, Edelstein, D, Geschwind, M, Brownlee, M, Federoff, HJ & Bergold, PJ 1993, 'Localized gene transfer into organotypic hippocampal slice cultures and acute hippocampal slices', Journal of Neuroscience Methods, vol. 50, no. 3, pp. 341-351. https://doi.org/10.1016/0165-0270(93)90040-X
Casaccia-Bonnefil, Patrizia ; Benedikz, Eirikur ; Shen, Hong ; Stelzer, Armin ; Edelstein, Diane ; Geschwind, Michael ; Brownlee, Michael ; Federoff, Howard J. ; Bergold, Peter J. / Localized gene transfer into organotypic hippocampal slice cultures and acute hippocampal slices. In: Journal of Neuroscience Methods. 1993 ; Vol. 50, No. 3. pp. 341-351.
@article{bfd37798458c4be28e472d3024c9030f,
title = "Localized gene transfer into organotypic hippocampal slice cultures and acute hippocampal slices",
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.",
keywords = "beta-Galactosidase, Gene transduction, Genetic vector, Herpes simplex virus-1, Microinjection",
author = "Patrizia Casaccia-Bonnefil and Eirikur Benedikz and Hong Shen and Armin Stelzer and Diane Edelstein and Michael Geschwind and Michael Brownlee and Federoff, {Howard J.} and Bergold, {Peter J.}",
year = "1993",
doi = "10.1016/0165-0270(93)90040-X",
language = "English (US)",
volume = "50",
pages = "341--351",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
number = "3",

}

TY - JOUR

T1 - Localized gene transfer into organotypic hippocampal slice cultures and acute hippocampal slices

AU - Casaccia-Bonnefil, Patrizia

AU - Benedikz, Eirikur

AU - Shen, Hong

AU - Stelzer, Armin

AU - Edelstein, Diane

AU - Geschwind, Michael

AU - Brownlee, Michael

AU - Federoff, Howard J.

AU - Bergold, Peter J.

PY - 1993

Y1 - 1993

N2 - 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.

AB - 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.

KW - beta-Galactosidase

KW - Gene transduction

KW - Genetic vector

KW - Herpes simplex virus-1

KW - Microinjection

UR - http://www.scopus.com/inward/record.url?scp=0027722973&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027722973&partnerID=8YFLogxK

U2 - 10.1016/0165-0270(93)90040-X

DO - 10.1016/0165-0270(93)90040-X

M3 - Article

C2 - 8152244

AN - SCOPUS:0027722973

VL - 50

SP - 341

EP - 351

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

IS - 3

ER -