Fig. 2: Inducible Col1a1-CreERT2 transgenic mice target perivascular fibroblasts in the uninjured mouse spinal cord. | Nature Neuroscience

Fig. 2: Inducible Col1a1-CreERT2 transgenic mice target perivascular fibroblasts in the uninjured mouse spinal cord.

From: Distinct origin and region-dependent contribution of stromal fibroblasts to fibrosis following traumatic injury in mice

Fig. 2

a, Col1a1+ perivascular cells can be labeled by tamoxifen-mediated genetic recombination in adult Col1a1-CreERT2 transgenic mice carrying R26R-tdTomato reporter alleles, in combination with the Pdgfrb-eGFP reporter line. b, Detection of Col1a1 mRNA in tdTomato+ cells in the uninjured spinal cord of Col1a1-CreERT2;R26R-tdTomato mice. c, Quantification of the average number of tdTomato+ cells per transverse spinal cord section (20-μm-thick) in uninjured Col1a1-CreERT2;R26R-tdTomato (mean ± s.d., 6.9 ± 1.6) and GLAST-CreERT2;R26R-tdTomato (mean ± s.d., 28.5 ± 6.6) mice. d,e, In the uninjured spinal cord, Col1a1-CreERT2 tdTomato+ cells express Pdgfrb-EGFP (white arrow) (d) and surround the blood vessel wall (marked with podocalyxin, white arrowhead) together with Pdgfrb-EGFP+ mural cells (yellow arrow) (e). f, Col1a1-CreERT2 tdTomato+ cells coexpress Col1a1 and the fibroblast marker Pdgfra (white arrows). The Pdgfra expression level in tdTomato+Col1a1+ fibroblasts is lower than in neighboring tdTomato−Col1a1−;Pdgfra+ cells (white arrowhead), which are presumably oligodendrocyte precursor cells. g, Slc1a3 (GLAST) is expressed by tdTomato+EGFP+ perivascular fibroblasts (white arrows) in the spinal cord of Col1a1-CreERT2;R26R-tdTomato;Pdgfrb-eGFP mice. In addition, Slc1a3 expression is detected in tdTomato−EGFP− cells (white arrowheads), which are mostly astrocytes, as well as in tdTomato−EGFP+ perivascular cells (yellow arrows). The insets show the close-up view of a tdTomato+EGFP+Slc1a3+ perivascular fibroblast. Pdx, podocalyxin; WM, white matter; GM, gray matter; tdTom, tdTomato. Scale bars: 100 μm (d), 20 μm (f, g), 10 μm (e, b) and 5 μm (insets in g). Data are shown as mean ± s.d. n = 5 (GLAST-CreERT2) and n = 7 (Col1a1-CreERT2) animals in c. ****P < 0.0001 by two-sided, unpaired Student’s t test in c. Dashed lines in d outline the spinal cord gray matter. Cell nuclei are labeled with 4′,6′-diamidino-2-phenylindole dihydrochloride (DAPI). Images are representative of two independent experiments. All images show transverse sections. Source data are provided as a source data file.

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