Wnt/β-Catenin Signaling Promotes Differentiation of Ischemia-Activated Adult Neural Stem/Progenitor Cells to Neuronal Precursors

Jan Kriska; Lucie Janeckova; Denisa Kirdajova; Pavel Honsa; Tomas Knotek; David Dzamba; Denisa Kolenicova; Olena Butenko; Martina Vojtechova; Martin Capek; Zbynek Kozmik; Makoto Mark Taketo; Vladimir Korinek; Miroslava Anderova

doi:10.3389/fnins.2021.628983

Wnt/β-Catenin Signaling Promotes Differentiation of Ischemia-Activated Adult Neural Stem/Progenitor Cells to Neuronal Precursors

Front Neurosci. 2021 Feb 25:15:628983. doi: 10.3389/fnins.2021.628983. eCollection 2021.

Authors

Jan Kriska¹, Lucie Janeckova², Denisa Kirdajova^{1

3}, Pavel Honsa¹, Tomas Knotek^{1

3}, David Dzamba¹, Denisa Kolenicova^{1

3}, Olena Butenko¹, Martina Vojtechova², Martin Capek⁴, Zbynek Kozmik⁵, Makoto Mark Taketo⁶, Vladimir Korinek², Miroslava Anderova^{1

3}

Affiliations

¹ Department of Cellular Neurophysiology, Institute of Experimental Medicine, Czech Academy of Sciences, Prague, Czechia.
² Laboratory of Cell and Developmental Biology, Institute of Molecular Genetics, Czech Academy of Sciences, Prague, Czechia.
³ Second Faculty of Medicine, Charles University, Prague, Czechia.
⁴ Service Laboratory of Light Microscopy, Institute of Molecular Genetics, Czech Academy of Sciences, Prague, Czechia.
⁵ Laboratory of Transcriptional Regulation, Institute of Molecular Genetics, Czech Academy of Sciences, Prague, Czechia.
⁶ Division of Experimental Therapeutics, Graduate School of Medicine, Kyoto University, Kyoto, Japan.

Abstract

Modulating endogenous regenerative processes may represent a suitable treatment for central nervous system (CNS) injuries, such as stroke or trauma. Neural stem/progenitor cells (NS/PCs), which naturally reside in the subventricular zone (SVZ) of the adult brain, proliferate and differentiate to other cell types, and therefore may compensate the negative consequences of ischemic injury. The fate of NS/PCs in the developing brain is largely influenced by Wingless/Integrated (Wnt) signaling; however, its role in the differentiation of adult NS/PCs under ischemic conditions is still enigmatic. In our previous study, we identified the Wnt/β-catenin signaling pathway as a factor promoting neurogenesis at the expense of gliogenesis in neonatal mice. In this study, we used adult transgenic mice in order to assess the impact of the canonical Wnt pathway modulation (inhibition or hyper-activation) on NS/PCs derived from the SVZ, and combined it with the middle cerebral artery occlusion (MCAO) to disclose the effect of focal cerebral ischemia (FCI). Based on the electrophysiological properties of cultured cells, we first identified three cell types that represented in vitro differentiated NS/PCs - astrocytes, neuron-like cells, and precursor cells. Following FCI, we detected fewer neuron-like cells after Wnt signaling inhibition. Furthermore, the immunohistochemical analysis revealed an overall higher expression of cell-type-specific proteins after FCI, indicating increased proliferation and differentiation rates of NS/PCs in the SVZ. Remarkably, Wnt signaling hyper-activation increased the abundance of proliferating and neuron-like cells, while Wnt pathway inhibition had the opposite effect. Finally, the expression profiling at the single cell level revealed an increased proportion of neural stem cells and neuroblasts after FCI. These observations indicate that Wnt signaling enhances NS/PCs-based regeneration in the adult mouse brain following FCI, and supports neuronal differentiation in the SVZ.

Keywords: Wnt signaling; adult neurogenesis; focal cerebral ischemia; gliogenesis; neural stem/progenitor cell; patch-clamp technique; single-cell RNA sequencing; transgenic mouse.