Abstract
The retina is an excellent model system for studies of neural development and disease due to its simple structure and accessibility. We have been using an electroporation technique to analyze gene structure and function rapidly and conveniently in the mouse and rat retinas in vivo and in vitro (in retinal explants). By electroporation, various types of DNA constructs are readily introduced into the retina without DNA size limitation. In addition, more than two different DNA constructs can be introduced into the same cells at once with very high co-transfection efficiency. With this technique, we have established protocols for inducible gene misexpression and knockdown, as well as conventional gene misexpression and knockdown, in the retina. These methods will be useful to reveal the molecular mechanisms of retinal development and disease.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Price J, Turner D, Cepko C (1987) Lineage analysis in the vertebrate nervous system by retrovirus-mediated gene transfer. Proc Natl Acad Sci U S A 84:156–160
Turner DL, Cepko CL (1987) A common progenitor for neurons and glia persists in rat retina late in development. Nature 328:131–136
Miyoshi H et al (1997) Stable and efficient gene transfer into the retina using an HIV-based lentiviral vector. Proc Natl Acad Sci U S A 94:10319–10323
Bennett J et al (1994) Adenovirus vector-mediated in vivo gene transfer into adult murine retina. Invest Ophthalmol Vis Sci 35:2535–2542
Li T et al (1994) In vivo transfer of a reporter gene to the retina mediated by an adenoviral vector. Invest Ophthalmol Vis Sci 35:2543–2549
Jomary C et al (1994) Adenovirus-mediated gene transfer to murine retinal cells in vitro and in vivo. FEBS Lett 347:117–122
Lamartina S et al (2007) Helper-dependent adenovirus for the gene therapy of proliferative retinopathies: stable gene transfer, regulated gene expression and therapeutic efficacy. J Gene Med 9:862–874
Ali RR et al (1996) Gene transfer into the mouse retina mediated by an adeno-associated viral vector. Hum Mol Genet 5:591–594
Chalberg TW et al (2005) PhiC31 integrase confers genomic integration and long-term transgene expression in rat retina. Invest Ophthalmol Vis Sci 46:2140–2146
Sato Y et al (2007) Stable integration and conditional expression of electroporated transgenes in chicken embryos. Dev Biol 305:616–624
Matsuda T, Cepko CL (2004) Electroporation and RNA interference in the rodent retina in vivo and in vitro. Proc Natl Acad Sci U S A 101:16–22
Matsuda T, Cepko CL (2007) Controlled expression of transgenes introduced by in vivo electroporation. Proc Natl Acad Sci U S A 104:1027–1032
Johnson CJ et al (2008) Technical brief: subretinal injection and electroporation into adult mouse eyes. Mol Vis 14:2211–2226
Kachi S et al (2005) Nonviral ocular gene transfer. Gene Ther 12:843–851
Dezawa M et al (2002) Gene transfer into retinal ganglion cells by in vivo electroporation: a new approach. Micron 33:1–6
Huberman AD et al (2005) Ephrin-As mediate targeting of eye-specific projections to the lateral geniculate nucleus. Nat Neurosci 8:1013–1021
Garcia-Frigola C et al (2007) Gene delivery into mouse retinal ganglion cells by in utero electroporation. BMC Dev Biol 7:103
Punzo C, Cepko CL (2008) Ultrasound-guided in utero injections allow studies of the development and function of the eye. Dev Dyn 237:1034–1042
Petros TJ, Shrestha BR, Mason C (2009) Specificity and sufficiency of EphB1 in driving the ipsilateral retinal projection. J Neurosci 29:3463–3474
Turner DL, Snyder EY, Cepko CL (1990) Lineage-independent determination of cell type in the embryonic mouse retina. Neuron 4:833–845
Anastassiadis K et al (2010) A practical summary of site-specific recombination, conditional mutagenesis, and tamoxifen induction of CreERT2. Methods Enzymol 477:109–123
Picard D (1994) Regulation of protein function through expression of chimaeric proteins. Curr Opin Biotechnol 5:511–515
Allocca M et al (2007) Novel adeno-associated virus serotypes efficiently transduce murine photoreceptors. J Virol 81:11372–11380
Niwa H, Yamamura K, Miyazaki J (1991) Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108:193–199
Schorpp M et al (1996) The human ubiquitin C promoter directs high ubiquitous expression of transgenes in mice. Nucleic Acids Res 24:1787–1788
Caffé AR et al (2001) Mouse retina explants after long-term culture in serum free medium. J Chem Neuroanat 22:263–273
Johnson TV, Martin KR (2008) Development and characterization of an adult retinal explant organotypic tissue culture system as an in vitro intraocular stem cell transplantation model. Invest Ophthalmol Vis Sci 49:3503–3512
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Matsuda, T. (2015). Electroporation in the Rodent Retina In Vivo and In Vitro. In: Saito, T. (eds) Electroporation Methods in Neuroscience. Neuromethods, vol 102. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2459-2_4
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2459-2_4
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2458-5
Online ISBN: 978-1-4939-2459-2
eBook Packages: Springer Protocols