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. 2011 Jul 29;16(4):515. doi: 10.2478/s11658-011-0021-0

Quantitative and kinetic profile of Wnt/β-catenin signaling components during human neural progenitor cell differentiation

Orianne Mazemondet 1,4, Rayk Hubner 1, Jana Frahm 1, Dirk Koczan 2, Benjamin M Bader 3, Dieter G Weiss 3, Adelinde M Uhrmacher 4, Moritz J Frech 1, Arndt Rolfs 1,✉, Jiankai Luo 1,✉
PMCID: PMC6275579  PMID: 21805133

Abstract

ReNcell VM is an immortalized human neural progenitor cell line with the ability to differentiate in vitro into astrocytes and neurons, in which the Wnt/β-catenin pathway is known to be involved. However, little is known about kinetic changes of this pathway in human neural progenitor cell differentiation. In the present study, we provide a quantitative profile of Wnt/β-catenin pathway dynamics showing its spatio-temporal regulation during ReNcell VM cell differentiation. We show first that T-cell factor dependent transcription can be activated by stabilized β-catenin. Furthermore, endogenous Wnt ligands, pathway receptors and signaling molecules are temporally controlled, demonstrating changes related to differentiation stages. During the first three hours of differentiation the signaling molecules LRP6, Dvl2 and β-catenin are spatio-temporally regulated between distinct cellular compartments. From 24 h onward, components of the Wnt/β-catenin pathway are strongly activated and regulated as shown by mRNA up-regulation of Wnt ligands (Wnt5a and Wnt7a), receptors including Frizzled-2, -3, -6, -7, and -9, and co-receptors, and target genes including Axin2. This detailed temporal profile of the Wnt/β-catenin pathway is a first step to understand, control and to orientate, in vitro, human neural progenitor cell differentiation.

Key words: Wnt/β-catenin pathway, Spatio-temporal dynamics, Quantitative kinetics

Full Text

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Abbreviations used

APC

adenomatous polyposis coli

bFGF

basic fibroblast growth factor

Cdk

cyclin-dependent kinase

CK1

casein kinase 1

DAPI

4′,6-diamidino-2-phenylindole

Dkk1

Dickkopf 1

DMEM

Dulbecco’s modified Eagle’s medium

Dvl

dishevelled

EGF

epidermal growth factor

Fz

Frizzled

GAPDH

glyceraldehyde 3-phosphate dehydrogenase

GFP

green fluorescent protein

GSK3beta

glycogen synthase kinase 3

HBSS

Hank’s buffered salt solution

hNPC

human neural progenitor cell

LRP6

low-density lipoprotein receptor-related protein 6

MAP2

microtubuleassociated protein 2

NPC

neural progenitor cell

Ror2

receptor tyrosine kinase-like orphan receptor 2

Ryk

receptor-like tyrosine kinase

TCF

T-cell factor

Footnotes

Both authors contributed equally to this work and should be considered co-first authors

Contributor Information

Arndt Rolfs, Phone: 0049-3814949540, FAX: 0049-3814949542, Email: arndt.rolfs@med.uni-rostock.de.

Jiankai Luo, Phone: 0049-3814949629, FAX: 0049-3814944899, Email: jiankai.luo@unirostock.de.

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