Injury-induced perivascular niche supports alternative differentiation of adult rodent CNS progenitor cells
- Justyna Ulanska-Poutanen
- Jakub Mieczkowski
- Chao Zhao
- Katarzyna Konarzewska
- Beata Kaza
- Lukasz Bugajski
- Bozena Kaminska
- Robin JM Franklin
- Malgorzata Zawadzka
- Nencki Institute of Experimental Biology, Polish Academy of Sciences, Poland
- University of Cambridge, United Kingdom
- Institute of Molecular Health Sciences, Switzerland
Figures
Figure 1 with 2 supplements
The cellular composition of VN differs from non-VN during remyelination.
(A) Schematic description of OPCs response to focal demyelination in rat CCP. Note the well-defined phases: recruitment of OPCs, their proliferation and differentiation (6, 10 and 14 dpl). Images …
Figure 1—figure supplement 1
Characteristics of cells occupied vascular niche area in remyelination time course.
(A) Density of OPCs positive for active caspase three in VN is similar to their density in non-VN during early remyelination (n = 3, mean ± SEM). (B) Identification of genetically labelled PDGFRA+ OP…
Figure 1—figure supplement 2
SCs-mediated remyelination is restricted to the vascular niche.
(A) Immunodetection for vWF+ blood vessels within the lesioned area (scale bar 100 μm). (B) SCs preferentially occupy discrete areas where astrocytes were absent even around the same vessel (high …
Figure 2 with 1 supplement
mRNA profiles differ in isolated microenvironmental niches.
(A) Images illustrate the microdissection procedure: areas around blood vessels marked to be cut out by UV-laser (VN) and areas of lesion without blood vessels marked to be cut afterwards (non-VN) …
-
Figure 2—source data 1
Genes significantly regulated in the non-vascular areas.
Dots represent level of expression for individual animal sample.
- https://doi.org/10.7554/eLife.30325.007
-
Figure 2—source data 2
Genes significantly regulated in the vascular niche.
Dots represent level of expression for individual animal sample.
- https://doi.org/10.7554/eLife.30325.008
-
Figure 2—source data 3
Genes significantly regulated only in the vascular niche.
Dots represent level of expression for individual animal sample.
- https://doi.org/10.7554/eLife.30325.009
Figure 2—figure supplement 1
Microarray analysis restricted to defined and putative endothelial and astrocyte enriched transcripts shows the different profiles of both niches in 10 dpl group.
For each individual gene average a ratio between average expression in vascular and non-vascular niche were computed. Expression ratios were plotted on a log2 scale. Gene signatures for …
Figure 3 with 1 supplement
Injury-induced vascular niche is enriched with BMP and Wnt morphogen transcripts.
(A) Graphical representation of BMP/WNT/Hedgehog signalling pathway interactions based on 10 dpl microarray results, higher expression of particular gene in non-VN in red, a higher expression in a …
Figure 3—figure supplement 1
BMP4, but not BMP2, is upregulated within the demyelinated lesion, regardless of the specific area examined.
Absolute expression of BMP4 and BMP2 in VN (green) and non-VN (red) at 6 and 10 dpl, dots represent expression level for each sample.
Figure 4 with 2 supplements
Spatiotemporal distribution and cellular sources of selected proteins.
(A) BMP4 transcript is produced in VN at 10 dpl (upper panel, in situ hybridization, scale bar 200 μm) mainly by OPCs (lower panel, Olig2 in red, scale bar 10 μm). (B) Sostdc1 transcript detection …
Figure 4—figure supplement 1
Olig2-positive cells producing Sostdc1, likely pre-myelinating OLs, are located close to the lesion edge.
Representative confocal images of 14 dpl lesion, the core of the lesion depicted by dotted line; NAWM, normal appeared white matter, scale bar 50 μm.
Figure 4—figure supplement 2
Gene expression analysis in the purified brain-derived cell populations at basal and activated conditions in vitro confirms the cellular origin of BMP4, Wnt2b, and Sostdc1.
(A) For in vitro expression analysis we used extra-pure populations of OPCs and astrocytes obtained by three times repeated shaking step since standard procedure of isolation results with astrocyte …
Figure 5 with 1 supplement
OPC differentiation fate at 4 weeks after transplantation into injured sciatic nerve.
(A) Gene expression pattern after sciatic nerve crush (mean ± sd; n = 3, Mann-Whithney U-test, *p<0.01 compared with 30 min after crush, #p<0.05 compared with 3 dpc). (B) Quantification of GFP+ cell …
Figure 5—figure supplement 1
OPCs differentiate abnormally when grafted into sciatic nerve at 3 or 6 dpc.
(A) GFP+ cells neither express periaxin (boxed area I reimaged using confocal microscope, right panel) nor GFAP and Olig2 (boxed area II, lower panel) when grafted 3 dpc, indicating their aberrant …
Figure 6
shRNA-mediated knockdown of LRP6 expression inhibits Schwann cell differentiation within injured sciatic nerve.
(A) Images show morphology of GFP+ OPCs transduced in vitro with lentiviral-mediated control and LRP6-specific shRNA, scale bar 20 μm (B) FACS sorting strategy, numbers show percent of cells of a …
Figure 7
Expression and accumulation of CTGF is restricted to the astrocytic response to demyelination.
(A) Expression of ECM proteins is niche-specific as revealed by microarray analysis (Fisher’s exact test, adj.pv <0.05). (B) CTGF expression decreases during remyelination; however it is …
Author response image 1
Graphs show absolute expression of BMP4 and BMP2 in VN (green) and non-VN (red) at 6 and 10 dpl, dots represent expression level for each sample.
The Y axis represents gene expression, log2; X axis – dpl.
Additional files
-
Transparent reporting form
- https://doi.org/10.7554/eLife.30325.019
