Abstract
Brain growth reflects the proliferation dynamics of neural progenitors, and understanding brain growth requires molecular, genetic, and functional studies of these specific cells. Cerebellar granule neuron progenitors (CGNPs) proliferate in the early postnatal period in both mice and humans, to generate the largest population of neurons in the central nervous system. CGNPs present a large, spatially segregated source of neural progenitors with a consistent, well-characterized temporal pattern of proliferation and differentiation that facilitates analysis. Dissociating of CGNPs with the methods below will generate a suspension of primary neural progenitors harvested from the postnatal brain that may be used for diverse experimental analyses including cell culture, protein extraction, flow cytometry, metabolomic analysis, and transcriptomic analysis with single-cell resolution (scRNA-seq).
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References
Ten Donkelaar HJ, Lammens M (2009) Development of the human cerebellum and its disorders. Clin Perinatol 36:513–530
Marzban H, Del Bigio MR, Alizadeh J et al (2015) Cellular commitment in the developing cerebellum. Front Cell Neurosci 8:1–26
Wechsler-Reya RJ, Scott MP (1999) Control of neuronal precursor proliferation in the cerebellum by Sonic Hedgehog. Neuron 22:103–114
Dahmane N, Ruiz i Altaba A (1999) Sonic hedgehog regulates the growth and patterning of the cerebellum. Development 126:3089–3100
Chizhikov V, Millen KJ (2003) Development and malformations of the cerebellum in mice. Mol Genet Metab 80:54–65
Garel C, Fallet-Bianco C, Guibaud L (2011) The fetal cerebellum: development and common malformations. J Child Neurol 26:1483–1492
Goldowitz D, Hamre K (1998) The cells and molecules that make a cerebellum. Trends Neurosci 21:375–382
Kenney AM (2003) Nmyc upregulation by sonic hedgehog signaling promotes proliferation in developing cerebellar granule neuron precursors. Development 130:15–28
Wallace VA (1999) Purkinje-cell-derived Sonic hedgehog regulates granule neuron precursor cell proliferation in the developing mouse cerebellum. Curr Biol 9:445–448
Solecki DJ, Liu X, Tomoda T et al (2001) Activated Notch2 signaling inhibits differentiation of cerebellar granule neuron precursors by maintaining proliferation. Neuron 31:557–568
Lang PY, Nanjangud GJ, Sokolsky-Papkov M et al (2016) ATR maintains chromosomal integrity during postnatal cerebellar neurogenesis and is required for medulloblastoma formation. Development 143:4038–4052
Krämer D, Minichiello L (2010) Mouse cell. Culture 633:233–239
Lee HY, Greene LA, Mason CA et al (2009) Isolation and culture of post-natal mouse cerebellar granule neuron progenitor cells and neurons. J Vis Exp 23:e990
Hatten ME (1985) Neuronal regulation of astroglial morphology and proliferation in vitro. J Cell Biol 100:384–396
Hatten ME, Gao W-Q, Morrison ME et al (1998) The cerebellum: purification and co-culture of identified cell populations. Cult Nerve Cells. G. Banker and K. Goslin, editors. MIT Press, Cambridge, MA, pp 419–459. https://cognet.mit.edu/book/culturing-nerve-cells
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Ocasio, J.K. (2023). Dissociation of Cerebellar Granule Neuron Progenitors for Culture, FACS, Transcriptomics, and Molecular Biology. In: Gershon, T. (eds) Microcephaly. Methods in Molecular Biology, vol 2583. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2752-5_1
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DOI: https://doi.org/10.1007/978-1-0716-2752-5_1
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