Abstract
Cortical development requires a strict balance between neuronal proliferation, differentiation, and cellular migration within restricted developmental stages. The precise spatiotemporal gene manipulation used in developmental studies can be achieved by in vitro or ex vivo experiments or by the generation of transgenic animals. However, these approaches have significant limitations when trying to investigate the origin and molecular regulation of early cortical neurons. In utero electroporation (IUE) is an informative cell labeling technique that provides the ability to label neural progenitor cells and their progeny in vivo with promoter-specific reporter constructs as well as to induce or repress gene expression in a spatially and temporally specific manner. Technical improvements of this method have allowed the targeting of multiple neural cell types, as well as the precise transfection of subpopulations of neurons at increasingly earlier embryonic stages. Furthermore, neuronal projection studies and the use of multiple electroporations in the same embryo have made it possible to examine processes occurring at different developmental stages and/or cortical areas and link their anatomy to their function. In this chapter, we present the latest advances of the in utero electroporation technique for the study of early formation of the cerebral cortex, together with a description of the necessary tools.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Monuki ES, Porter FD, Walsh CA (2001) Patterning of the dorsal telencephalon and cerebral cortex by a roof plate-Lhx2 pathway. Neuron 32:591–604
O’Leary DDM, Chou S-J, Sahara S (2007) Area patterning of the mammalian cortex. Neuron 56:252–269
Borello U, Pierani A (2010) Patterning the cerebral cortex: traveling with morphogens. Curr Opin Genet Dev 20:408–415
Caronia-Brown G, Yoshida M, Gulden F et al (2014) The cortical hem regulates the size and patterning of neocortex. Development 141:2855–2865
Chédotal A, Richards LJ (2010) Wiring the brain: the biology of neuronal guidance. Cold Spring Harb Perspect Biol 2:a001917
Shimogori T, Grove EA (2005) Fibroblast growth factor 8 regulates neocortical guidance of area-specific thalamic innervation. J Neurosci 25:6550–6560
Assimacopoulos S, Kao T, Issa NP, Grove EA (2012) Fibroblast growth factor 8 organizes the neocortical area map and regulates sensory map topography. J Neurosci 32:7191–7201
Saito T, Nakatsuji N (2001) Efficient gene transfer into the embryonic mouse brain using in vivo electroporation. Dev Biol 240:237–246
Tabata H, Nakajima K (2001) Efficient in utero gene transfer to the developing mouse brain using electroporation: visualization of neuronal migration in the developing cortex. Neuroscience 103:865–872
Britanova O, De Juan RC, Cheung A et al (2008) Satb2 is a postmitotic determinant for upper-layer neuron specification in the neocortex. Neuron 57:378–392
Shibata M, Kurokawa D, Nakao H et al (2008) MicroRNA-9 modulates Cajal-Retzius cell differentiation by suppressing foxg1 expression in mouse medial pallium. J Neurosci 28:10415–10421
Rouaux C, Arlotta P (2013) Direct lineage reprogramming of post-mitotic callosal neurons into corticofugal neurons in vivo. Nat Cell Biol 15:214–221
Srivatsa S, Parthasarathy S, Britanova O et al (2014) Unc5C and DCC act downstream of Ctip2 and Satb2 and contribute to corpus callosum formation. Nat Commun 5:1–15
Faux C, Rakic S, Andrews W, Britto JM (2012) Neurons on the move: migration and lamination of cortical interneurons. Neurosignals 20:168–189
Borrell V, Yoshimura Y, Callaway EM (2005) Targeted gene delivery to telencephalic inhibitory neurons by directional in utero electroporation. J Neurosci Methods 143:151–158
Takiguchi-Hayashi K, Sekiguchi M, Ashigaki S et al (2004) Generation of reelin-positive marginal zone cells from the caudomedial wall of telencephalic vesicles. J Neurosci 24:2286–2295
Bielle F, Griveau A, Narboux-Nême N et al (2005) Multiple origins of Cajal-Retzius cells at the borders of the developing pallium. Nat Neurosci 8:1002–1012
Huilgol D, Udin S, Shimogori T et al (2013) Dual origins of the mammalian accessory olfactory bulb revealed by an evolutionarily conserved migratory stream. Nat Neurosci 16:157–165
Remedios R, Huilgol D, Saha B et al (2007) A stream of cells migrating from the caudal telencephalon reveals a link between the amygdala and neocortex. Nat Neurosci 10:1141–1150
Suárez R, Fenlon LR, Marek R et al (2014) Balanced interhemispheric cortical activity is required for correct targeting of the corpus callosum. Neuron 82:1289–1298
Zhou J, Wen Y, She L et al (2013) Axon position within the corpus callosum determines contralateral cortical projection. Proc Natl Acad Sci U S A 110:E2714–E2723
Sahara S, Kawakami Y, Izpisua Belmonte J, O’Leary DD (2007) Sp8 exhibits reciprocal induction with Fgf8 but has an opposing effect on anterior-posterior cortical area patterning. Neural Dev 2:1–22
Yaguchi M, Ohashi Y, Tsubota T et al (2013) Characterization of the properties of seven promoters in the motor cortex of rats and monkeys after lentiviral vector-mediated gene transfer. Hum Gene Ther Methods 24:333–344
Yoshida A, Yamaguchi Y, Nonomura K et al (2010) Simultaneous expression of different transgenes in neurons and glia by combining in utero electroporation with the Tol2 transposon-mediated gene transfer system. Genes Cells 15:501–512
Chen F, Maher BJ, LoTurco JJ (2014) PiggyBac transposon-mediated cellular transgenesis in mammalian forebrain by in utero electroporation. Cold Spring Harb Protoc 2014: 741–749
Loulier K, Barry R, Mahou P et al (2014) Multiplex cell and lineage tracking with combinatorial labels. Neuron 81:505–520
Acknowledgements
We thank R. Suárez and L. Fenlon for their input on the manuscript and providing histological images; Queensland Brain Institute (QBI) Microscopy Facility for microscopy assistance; the University of Queensland Biological Resources and QBI animal team for animal support. PK was supported by a National Health and Medical Research Council (NHMRC, Australia) Early Career (CJ Martin) Fellowship, GA was supported by a Becas Chile Fellowship, and LJR was supported by an NHMRC Principal Research Fellowship. IG was supported by NHMRC project grant GNT1048849.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kozulin, P., Almarza, G., Gobius, I., Richards, L.J. (2016). Investigating Early Formation of the Cerebral Cortex by In Utero Electroporation: Methods and Protocols. In: Walker, D. (eds) Prenatal and Postnatal Determinants of Development. Neuromethods, vol 109. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3014-2_1
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3014-2_1
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3013-5
Online ISBN: 978-1-4939-3014-2
eBook Packages: Springer Protocols