Skip to main content

Manipulating Potassium Channel Expression and Function in Hippocampal Neurons by In Utero Electroporation

Part of the Methods in Molecular Biology book series (MIMB,volume 1684)

Abstract

In utero electroporation (IUE) of plasmid DNA into specific brain regions such as hippocampus and cortex has made it possible to reduce protein expression levels or even replace the endogenous protein with site-directed mutant proteins to reveal important physiological consequences. For example, shRNA can mediate targeted knockdown, and can be complemented by simultaneously expressing the shRNA immune wild-type protein to validate on-target effects, or by expression of an shRNA-immune protein harboring site-specific mutations. More recently, IUE has been adapted to express CRISPR/Cas9 components for targeted gene editing that abolishes protein expression. Utilizing these approaches via IUE results in transfected neurons that are interspersed with non-transfected control neurons in the same brain slices, allowing for direct comparisons. Because IUE is performed late in embryonic development and is confined to a relatively small percentage of neurons, developmental compensatory mechanisms that may compromise gene targeting via germ line transmission are minimized. Thus, IUE presents a powerful opportunity to rapidly and cost-effectively dissect molecular and cellular physiology in the context of the intact brain.

Key words

  • Potassium channel
  • In utero electroporation
  • Hippocampus
  • Neurons
  • shRNA
  • Animal model

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Saito T, Nakatsuji N (2001) Efficient gene transfer into the embryonic mouse brain using in vivo electroporation. Dev Biol 240:237–246

    CAS  CrossRef  PubMed  Google Scholar 

  2. Tabata H, Nakajima K (2001) Efficient in utero gene transfer system to the developing mouse brain using electroporation: visualization of neuronal migration in the developing cortex. Neuroscience 103:865–872

    CAS  CrossRef  PubMed  Google Scholar 

  3. Wang K, Lin MT, Adelman JP, Maylie J (2014) Distinct Ca2+ sources in dendritic spines of hippocampal CA1 neuron couple to SK and Kv4 channels. Neuron 81(2):379–387

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kang Wang .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

© 2018 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Wang, K. (2018). Manipulating Potassium Channel Expression and Function in Hippocampal Neurons by In Utero Electroporation. In: Shyng, SL., Valiyaveetil, F., Whorton, M. (eds) Potassium Channels. Methods in Molecular Biology, vol 1684. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7362-0_1

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7362-0_1

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7361-3

  • Online ISBN: 978-1-4939-7362-0

  • eBook Packages: Springer Protocols