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Genetic manipulation of adult mouse neurogenic niches by in vivo electroporation

Nature Methods volume 5, pages 189–196 (2008)Cite this article

Abstract

Targeted ectopic expression of genes in the adult brain is an invaluable approach for studying many biological processes. This can be accomplished by generating transgenic mice or by virally mediated gene transfer, but these methods are costly and labor intensive. We devised a rapid strategy that allows localized in vivo transfection of plasmid DNA within the adult neurogenic niches without detectable brain damage. Injection of plasmid DNA into the ventricular system or directly into the hippocampus of adult mice, followed by application of electrical current via external electrodes, resulted in transfection of neural stem or progenitor cells and mature neurons. We showed that this strategy can be used for both fate mapping and gain- or loss-of-function experiments. Using this approach, we identified an essential role for cadherins in maintaining the integrity of the lateral ventricle wall. Thus, in vivo electroporation provides a new approach to study the adult brain.

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Figure 1: Gene transfer to the adult brain without detectable damage by electroporation.
Figure 2: Cell types targeted by electroporation in the lateral ventricle wall.
Figure 3: Electroporation targets cells with neural stem cell properties.
Figure 4: Targeting the hippocampal neurogenic niche by electroporation.
Figure 5: Adult brain electroporation allows gain- and loss-of-function experiments.
Figure 6: Cadherins are essential for the integrity of the adult lateral ventricle wall.

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Acknowledgements

We thank R. Kageyama (Kyoto University) for the gift of the nestin promoter vector30, C. Ibáñez (Karolinska Institutet) for the gift of a BDNF expression plasmid, M. Wheelock (University of Nebraska Medical Center) for providing the dominant-negative N-cadherin cDNA and K. Fernandes (University of Montreal) for critically reading the manuscript. This study was supported by grants from the Swedish Research Council, the Swedish Cancer Society, the Foundation for Strategic Research, the Karolinska Institutet, Tobias Stiftelsen and the European Commission Framework VI Programme, EuroStemCell. F.B.-H. is supported by a postdoctoral fellowship from Canadian Institutes of Health Research.

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Author notes

  1. Konstantinos Meletis

    Present address: Present address: The Picower Institute for Learning and Memory, Massachusetts Institute of Technology, 43 Vassar Street, Cambridge, Massachusetts 02139, USA.,

Authors and Affiliations

  1. Department of Cell and Molecular Biology, Medical Nobel Institute, Stockholm, SE-171 77, Sweden

    Fanie Barnabé-Heider, Konstantinos Meletis, Malin Eriksson, Olaf Bergmann, Hanna Sabelström, Harald Mikkers & Jonas Frisén

  2. Department of Neuroscience, Box 285, Karolinska Institutet, Stockholm, SE-171 77, Sweden

    Michael A Harvey

  3. Department of Molecular Cell Biology and Regenerative Medicine Program, Leiden University Medical Center, P.O. Box 9600, RC Leiden, 2300, The Netherlands

    Harald Mikkers

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  1. Fanie Barnabé-Heider
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Contributions

F.B.-H. designed, performed and analyzed the study and wrote the manuscript; K.M. designed and performed most parts of the study; M.E. performed and analyzed part of the study (including BAC analysis); O.B. performed part of the study; H.S. performed and analyzed part of the study; M.A.H. performed EEG analysis; H.M. designed and performed part of the adenoviral study; and J.F. designed the study and wrote the manuscript.

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Correspondence to Jonas Frisén.

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Barnabé-Heider, F., Meletis, K., Eriksson, M. et al. Genetic manipulation of adult mouse neurogenic niches by in vivo electroporation. Nat Methods 5, 189–196 (2008). https://doi.org/10.1038/nmeth.1174

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