Fig. 5

The behavioral scores of rats with spinal cord injury (SCI) after injection of Zfp-IRES-GFP (Zfp-GFP) and pSFFV-IRES-GFP (Mock) lentiviral particles at 6 weeks post-transduction (WPT). a Schematic of the experimental procedure used for in vivo reprogramming and analysis over 7 weeks. The stereotaxic surgery was performed 1 week before SCI. The contusion model was created at week 1, and lentiviruses that contained Zfp-IRES-GFP or pSFFV-IRES-GFP were injected at 0 weeks post-transduction (WPT). Electrophysiological examinations were performed 2 days before SCI, 2 days after SCI, and at 6 WPT by MEP. d, day; w, week; MEP, motor evoked potential; SCI, spinal cord injury. b Zfp-IRES-GFP vector structure. The vector included an SFFV promoter, a ZFP521 CDS, and GFP that were linked by an IRES sequence. c Fluorescent and bright-field image of transduced astrocytes by Zfp-GFP vector at 1 WPT. d The immunofluorescent staining of transduced cells at 4 WPT for Dcx. Note that flat astrocytes were converted into Dcx⁺ bipolar cells. e The behavioral scores and open-field locomotor assessment as measured by the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale. The animals in intact (or normal, n = 6), Mock (n = 6), Zfp (n = 13), AST-Mock (n = 5), and AST-Zfp (n = 8) groups were assessed weekly for 7 weeks. The BBB analysis showed a significant improvement in the AST-Zfp group in comparison with the Mock and AST-Mock group and an even greater improvement in the Zfp group. f, g BBB scores were further analyzed by calculating subscores. BBB subscores represent measures of paw position, toe clearance, trunk control, and tail position made independent of all other observable traits. Analysis of BBB subscores showed significant improvement in paw position, toe clearance, trunk control, and tail position for the Zfp group at 6 WPT. Data in e and g are given as mean ± SD. Data were analyzed by ANOVA and Mann-Whitney U test as post hoc. *p < 0.05, **p < 0.01, and ***p < 0.001