Abstract
Transfection allows the introduction of foreign nucleic acid into eukaryotic cells. It is an important tool in understanding the roles of NMDARs in neurons. Here, we describe using lipofection-mediated transfection to introduce cDNA encoding NMDAR subunits into postmitotic rodent primary cortical neurons maintained in culture.
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References
Karra D, Dahm R (2010) Transfection techniques for neuronal cells. J Neurosci 30:6171–6177. doi:10.1523/JNEUROSCI.0183-10.2010
Zeitelhofer M, Vessey JP, Xie Y, Tübing F, Thomas S, Kiebler M et al (2007) High-efficiency transfection of mammalian neurons via nucleofection. Nat Protoc 2:1692–1704. doi:10.1038/nprot.2007.226
Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M et al (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A 84:7413–7417. doi:10.1073/pnas.84.21.7413
Felgner PL, Ringold GM (1989) Cationic liposome-mediated transfection. Nature 337(6205):387–388. doi:10.1038/337387a0
Bading H, Ginty DD, Greenberg ME (1993) Regulation of gene expression in hippocampal neurons by distinct calcium signaling pathways. Science 260:181–186. doi:10.1126/science.8097060
Papadia S, Soriano FX, Léveillé F, Martel M-A, Dakin KA, Hansen HH et al (2008) Synaptic NMDA receptor activity boosts intrinsic antioxidant defenses. Nat Neurosci 11:476–487. doi:10.1038/nn2071
Butash KA, Natarajan P, Young A, Fox DK (2000) Reexamination of the effect of endotoxin on cell proliferation and transfection efficiency. BioTechniques 29:610–619
Acknowledgments
This work was supported by a Wellcome Trust Clinical PhD studentship grant to KFMM. Much of the work put into optimizing this protocol was done by others in GEH’s lab over the last decade: particularly Sofia Papadia and Sean McKay. Michael Greenberg’s lab developed the principles of transfection in neurons.
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Marwick, K.F.M., Hardingham, G.E. (2017). Transfection in Primary Cultured Neuronal Cells. In: Burnashev, N., Szepetowski, P. (eds) NMDA Receptors. Methods in Molecular Biology, vol 1677. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7321-7_6
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DOI: https://doi.org/10.1007/978-1-4939-7321-7_6
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7321-7
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