Cell tracking in vitro reveals that the extracellular matrix glycoprotein Tenascin-C modulates cell cycle length and differentiation in neural stem/progenitor cells of the developing mouse spinal cord

Abstract

Generation of astrocytes during the development of the mammalian spinal cord is poorly understood. Previously, we have shown that the glycoprotein of the extracellular matrix (ECM) tenascin-C (Tnc) modulates the expression territories of the patterning genes Nkx6.1 and Nkx2.2 in the developing ventral spinal cord, tunes the responsiveness of neural stem/progenitor cells towards the cytokines FGF2 and EGF and thereby promotes astrocyte maturation. In order to obtain further mechanistic insight into these processes, we have compared embryonic day-15 spinal cord neural progenitor cells (NPCs) from wild-type and Tnc knockout mice using continuous single-cell live imaging and cell lineage analysis in vitro. Tnc knockout cells displayed a significantly reduced rate of cell division both in response to FGF2 and EGF. When individual clones of dividing cells were investigated with regard to their cell lineage trees using the tTt tracking software, it appeared that the cell cycle length in response to growth factors was reduced in the knockout. Furthermore, when Tnc knockout NPCs were induced to differentiate by the removal of FGF2 and EGF glial differentiation was enhanced. We conclude that the constituent of the stem cell niche Tnc contributes to preserve stemness of NPCs.

Sulfation of Glycosaminoglycans Modulates the Cell Cycle of Embryonic Mouse Spinal Cord Neural Stem Cells

“…At the same time, NSPCs isolated from the spinal cord of tenascin-C-deficient mice were analyzed in an analogous manner for an independent experimental approach. The dataset regarding the untreated control cells served as reference both for the chlorate-treated and the tenascin-C depleted NSPCs and has been published in the latter context ( May et al, 2018 ). Here, as the experiments have been conducted in parallel, this data set of the control cells was used as reference to evaluate the influence of inhibited sulfation on spinal cord-derived NSPCs.…”

“…Finally, full lineage trees originating from an individual stem cell could be constructed in that manner. A previous study of the laboratory illustrates this procedure, documenting the simultaneous construction of lineage trees in parallel with the time-lapse video ( May et al, 2018 ). Movies were created using ImageJ 1.45r (National Institutes of Health) software and are played at a speed of five frames per second (see Supplementary Material ).…”

“…In order to assess the potential functions of proteoglycans from the phosphacan/RPTP-β/ζ family we examined their expression and the presence of distinct CS-sulfotransferases during spinal cord development. To gain insight into the influence of sulfated GAGs on the cell cycle, we performed time-lapse video microscopy and single cell tracking of spinal cord progenitors treated with sodium chlorate, as a potent pharmacological sulfation inhibitor ( Eilken et al, 2009 ; Rieger et al, 2009 ; Sirko et al, 2010a ; Costa et al, 2011 ; Hoppe et al, 2016 ; May et al, 2018 ). Here, we show that the proliferation and lineage relationship of spinal cord-derived NSPCs cultivated in the presence of FGF2 are strongly altered by suppressing sulfation.…”

Advanced technological tools to study multidrug resistance in cancer

“…The presently available tools for single-cell tracking are missing methodology to resolve ambiguities such as joint multi-target tracking and classification (Hilsenbeck et al, 2016;Hormoz et al, 2016;Rapoport et al, 2011;Stadler et al, 2018). However, the development of software tracking tools is under constant improvement (Davis et al, 2007;Errington et al, 2005;Khan et al, 2007;Koh et al, 2017;May et al, 2018). The latest advancements appear to focus on automation based on localized image analysis and recognition.…”

Spidroin Silk Fibers with Bioactive Motifs of Extracellular Proteins for Neural Tissue Engineering

“… 19 , 20 For tenascin-C, it was shown that it modulates cell cycle length and differentiation in neural stem/progenitor cells and participates in the formation of the radial glia type B of the adult CNS stem cell niches. 21 , 22 However, it remains unclear how these proteins and their active peptides influence on drNPC differentiation.…”