Transfection of rat or mouse neurons by biolistics or electroporation

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Abstract

Properties of ion channels are affected by the background of the cells in which they are expressed. Thus, it is important for investigators interested in neuronal function to study these proteins in post-mitotic neurons. However, post-mitotic neurons, and many cell lines, are difficult to transfect by standard methods. Here we provide detailed protocols for two different procedures, biolistic and electroporation, which have been used to transfect peripheral sensory neurons from mice or rats with expression constructs of voltage-gated sodium channels. Neurons can be prepared, transfected and currents recorded within 48 h. Using these methods, primary sensory neurons can be transfected with an efficiency of 5–20%, which has permitted studying biophysical properties of sodium channels and their naturally occurring mutants in a native neuronal cell background. Although we have used sodium channels for the examples that we show here, these methods can also be used to study other types of molecules.

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Figure 1: Expression of recombinant Nav1.3 and Nav1.6 channels in Nav1.8-null mouse DRG neurons.
Figure 2: Expression of Nav1.8 channels in Nav1.8-null mouse DRG neurons.

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Acknowledgements

We thank Dr. Mark Estacion, Shujun Liu, Rachel Blackman, Bart Toftness and other members of S.G.W. laboratory for valuable assistance in development and refinement of the techniques described in this paper. Work in S.G.W. laboratory is supported in part by grants from the National Multiple Sclerosis Society and from the Rehabilitation Research and Development Service and Medical Research Service, Department of Veterans Affairs. T.R.C. was supported by research Grant NS053422 from the National Institutes of Health. The Center for Neuroscience and Regeneration Research is a collaboration of the Paralyzed Veterans of America and the United Spinal Association with Yale University.

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Correspondence to Stephen G Waxman.

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Dib-Hajj, S., Choi, J., Macala, L. et al. Transfection of rat or mouse neurons by biolistics or electroporation. Nat Protoc 4, 1118–1127 (2009) doi:10.1038/nprot.2009.90

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