Abstract
In the developing central nervous system (CNS), neural stem cells undergo a complex series of morphogenetic and motile events. Errors in neural stem cell proliferation or migration cause serious brain developmental disorders. However, the relative importance of each step in neurogenesis and migration and the identity of genes affecting these processes has only begun to be explored. Using live imaging in brain slices, neural stem cells and their progeny labeled by in utero gene transfer can be monitored at high spatial and temporal resolution for as long as several days. Cell cycle progression, mitosis, morphogenesis, and migratory behavior can each be documented in detail. Furthermore, the behavior of subcellular structures, including nuclei, centrosomes, and microtubules, can also be observed using fluorescent marker proteins. This chapter describes the application of these approaches in combination with RNA interference to investigate normal developing brain and the role of genes involved in brain developmental disorders, such as lissencephaly.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Malatesta, P., Hartfuss, E., and Gotz, M. (2000) Isolation of radial glial cells by fluorescent-activated cell sorting reveals a neuronal lineage Development 127, 5253–63
Miyata, T., Kawaguchi, A., Okano, H., and Ogawa, M (2001) Asymmetric inheritance of radial glial fibers by cortical neurons. Neuron 31, 727–41
Noctor, S. C., Flint, A. C., Weissman, T. A., Dammerman, R. S., and Kriegstein, A. R. (2001) Neurons derived from radial glial cells establish radial units in neocortex Nature 409, 714–20
Sauer, F. C. (1935) Mitosis in the neural tube J Comp Neurol 62, 377–405
Noctor, S. C., Martinez-Cerdeno, V., Ivic, L., and Kriegstein, A. R. (2004) Cortical neurons arise in symmetric and asymmetric division zones and migrate through specific phases Nat Neurosci 7, 136–44
Rakic, P., Stensas, L. J., Sayre, E., and Sidman, R. L. (1974) Computer-aided three-dimensional reconstruction and quantitative analysis of cells from serial electron microscopic montages of foetal monkey brain Nature 250, 31–4
Tabata, H., and Nakajima, K. (2003) Multipolar migration: the third mode of radial neuronal migration in the developing cerebral cortex J Neurosci 23, 9996–10001
Saito, T., and Nakatsuji, N. (2001) Efficient gene transfer into the embryonic mouse brain using in vivo electroporation Dev Biol 240, 237–46
Tabata, H., and 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, 865–72
Tsai, J. W., Chen, Y., Kriegstein, A. R., and Vallee, R. B. (2005) LIS1 RNA interference blocks neural stem cell division, morphogenesis, and motility at multiple stages J Cell Biol 170, 935–45
Tsai, J. W., Bremner, K. H., and Vallee, R. B. (2007) Dual subcellular roles for LIS1 and dynein in radial neuronal migration in live brain tissue Nat Neurosci 10, 970–9
Schaar, B. T., and McConnell, S. K. (2005) Cytoskeletal coordination during neuronal migration Proc Natl Acad Sci USA 102, 13652–7
McManus, M. F., Nasrallah, I. M., Pancoast, M. M., Wynshaw-Boris, A., and Golden, J. A. (2004) Lis1 is necessary for normal non-radial migration of inhibitory interneurons Am J Pathol 165, 775–84
Saito, T. (2006) In vivo electroporation in the embryonic mouse central nervous system Nat Protoc 1, 1552–8
Tsai, J. W., Lian, W. N., Kemal, S., Kriegstein, A. R., and Vallee, R. B. (2010) Kinesin 3 and cytoplasmic dynein mediate interkinetic nuclear migration in neural stem cells Nat Neurosci 13, 1463–71
Acknowledgments
The authors thank Dr. Wei-Nan Lian and Shahrnaz Kemal for critical reading of this manuscript. This work was supported by NIH grants HD40182 and GM47434, the March of Dimes Birth Defects Foundation to RBV, and the New York State Spinal Cord Injury Research Board to JWT.
Another application of the methods described in this article has recently been published by our lab (15).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Tsai, JW., Vallee, R.B. (2011). Live Microscopy of Neural Stem Cell Migration in Brain Slices. In: Filippi, MD., Geiger, H. (eds) Stem Cell Migration. Methods in Molecular Biology, vol 750. Humana Press. https://doi.org/10.1007/978-1-61779-145-1_9
Download citation
DOI: https://doi.org/10.1007/978-1-61779-145-1_9
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-144-4
Online ISBN: 978-1-61779-145-1
eBook Packages: Springer Protocols