Preservation of positional identity in fetus-derived neural stem (NS) cells from different mouse central nervous system compartments

Abstract

Neural stem (NS) cells are a self-renewing population of symmetrically dividing multipotent radial glia-like stem cells, characterized by homogeneous expansion in monolayer. Here we report that fetal NS cells isolated from different regions of the developing mouse nervous system behave in a similar manner with respect to self-renewal and neuropotency, but exhibit distinct positional identities. For example, NS cells from the neocortex maintain the expression of anterior transcription factors, including Otx2 and Foxg1, while Hoxb4 and Hoxb9 are uniquely found in spinal cord-derived NS cells. This molecular signature was stable for over 20 passages and was strictly linked to the developmental stage of the donor, because only NS cells derived from E14.5 cortex, and not those derived from E12.5 cortex, carried a consistent transcription factor profile. We also showed that traits of this positional code are maintained during neuronal differentiation, leading to the generation of electrophysiologically active neurons, even if they do not acquire a complete neurochemical identity.

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Abbreviations

BDNF:

Brain-derived neurotrophic factor

CNS:

Central nervous system

D-V:

Dorsoventral

EGF:

Epidermal growth factor

ESC:

Embryonic stem cell

FGF-2:

Fibroblast growth factor 2

GABA:

γ-Aminobutyric acid

GAP-43:

Growth-associated protein 43

GFAP:

Glial fibrillary acid protein

LGE:

Lateral ganglionic eminence

MAP2:

Microtubule-associated protein 2

MGE:

Medial ganglionic eminence

MN:

Motorneuron

NS cells:

Radial glia-like neural stem cells

R-C:

Rostrocaudal

Shh:

Sonic hedgehog

SVZ:

Subventricular zone

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Acknowledgments

This research was supported by initial funding from Fondazione Cariplo (Italy) in the context of the awarded N.O.B.E.L. (Operational Network for Biomedicine par Excellence in Lombardy) project entitled “A genetic toolkit for the analyses of neural stem cells—acronym: Mouse NS-toolkit”. This work is also supported by EuroSystem (FP7, European Union Health-F4-2008-200720), Neuroscreen (FP6, European Union LSHB-CT-2007-037766), and NeuroStemcell (FP7, European Union HEALTH-2008-B-222943). The laboratory also acknowledges the contribution of Unicredit Banca S.p.A. (Italy) and of Tavola Valdese (Italy).

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Correspondence to Elena Cattaneo.

Additional information

An erratum to this article is available at http://dx.doi.org/10.1007/s00018-016-2307-x.

Electronic supplementary material

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Figure S1. RT–PCR analysis of selected neural progenitor/stem cell markers (JPEG 750 kb)

Figure S2. Immunofluorescence characterization of E14.5 neocortex-derived NSCs. Self-renewing NS14CX cells homogeneously showed double immunoreactivity for Nestin/Vimentin and SOX2/PAX6 (JPEG 1804 kb)

Figure S3. Immunofluorescence characterization of 3 different clones of NS cells derived from E12.5 neocortex (NS12CX) and 3 clones of NS cells derived from E12.5 spinal cord (NS12SC). Self-renewing NS cell clones homogeneously express the typical NS cell markers Nestin, BLBP, RC2, Vimentin, and OLIG2 (JPEG 2075 kb)

Figure S4. Neuronal differentiation of NS12SC. This image shows a GAP-43-positive neuron derived from NS12SC cells differentiated for 23 d (JPEG 646 kb)

Figure S5. RT–PCR analysis of a subset of neurotransmitter receptors, described in striatal and spinal cord neurons in vivo. Neither proliferating (P) nor differentiating (D) NS cells expressed functional glutamatergic, glycinergic, or cholinergic receptors, even if the muscarinic receptor (Chrm4) was detectable (JPEG 579 kb)

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Onorati, M., Binetti, M., Conti, L. et al. Preservation of positional identity in fetus-derived neural stem (NS) cells from different mouse central nervous system compartments. Cell. Mol. Life Sci. 68, 1769–1783 (2011). https://doi.org/10.1007/s00018-010-0548-7

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Keywords

  • Neural stem cells
  • Neural development
  • Positional identity
  • Neuronal differentiation
  • Transcription factors