Fig. 2: In vivo application of ESAL-expressing NS/PC transplantation after cervical spinal cord injury.
From: A non-invasive system to monitor in vivo neural graft activity after spinal cord injury
a Schematic illustration of the positive control mice. Transplantation of NS/PCs double-infected with the two viruses (CAG-hM3Dq-mCherry, which contains an hM3Dq and mCherry fusion protein, and ESAL were transduced into NS/PCs via lentivirus) was performed 9 days after C5 dorsal column transection. Six weeks after transplantation, luminescence measurements were performed before sacrifice. b Comparison of the ratios of BLI signal intensity (7 h post-CNO/pre-CNO) between ESAL-expressing NS/PC-transplanted (ESAL-NS/PC-transplanted) mice (hM3Dq [−] TP, n = 4) and double-infected NS/PC-transplanted mice (hM3Dq [+] TP, n = 3) are shown. The values are the mean ± SEM: *p < 0.05. A two-sided unpaired Student’s t test was performed. T value and degrees of freedom: t(5) = 3.977, p = 0.011. c Representative IVIS images of a positive control mouse (pre- and post-CNO). The circle shows the region of interest (ROI) in the cervical spine. The colour of the bars indicates the total bioluminescence radiance (photons/sec/cm2/str). d, e Representative images from a positive control mouse 6 weeks after transplantation; labelled with Venus (green), mCherry (red), and HNA (human cells) (blue) (d) or labelled with Venus (green), Fos (red), and HNA (blue) (e). Scale bars, 20 μm. f Schematic illustration of in vivo experiments. Transplantation of ESAL-NS/PCs was performed 9 days after C5 dorsal column transection. Three, 6, and 9 weeks after transplantation, luminescence measurements were performed. All mice were sacrificed 10 weeks after transplantation. g Representative images of grafted cells from an ESAL-NS/PC-transplanted mouse labelled with Venus (green), pan-ELAVL (red; arrowheads), and HNA (blue). Scale bars, 20 μm. h Time-dependent change in graft luminescence intensity of ESAL-NS/PC-transplanted mice at 3, 6, and 9 weeks after transplantation (n = 12 mice). The values are the mean ± SEM: *p < 0.05. N.S.: not significant. A repeated-measures ANOVA was performed. Individual p values: weeks 3 and 6; 0.1735, weeks 6 and 9; 0.2007, weeks 3 and 9; 0.0035. i Representative images of grafted cells from ESAL-NS/PC-transplanted mouse tissues labelled with Venus (green), APC (oligodendrocytes), GFAP (astrocytes), Ki-67/Nestin (red; arrowheads), and HNA (blue). Scale bars, 20 μm.
