Figure 1.

Longitudinal evaluation of post-stroke recovery in mice receiving anti-CD49d treatment.(A) The lesion area based on autofluorescence after stroke of individual animals was superimposed to depict the lesion throughout the observation period of 84 d in Thy1-GCaMP6s animals. The color code indicates the sum of overlapping lesion pixels of individual mice per acquisition time point. Data were generated in two independent experiments. BL, baseline; D, day. (B) Quantification of the number of autofluorescent pixels seen in calcium imaging of Thy1-GCaMP6s animals. Control (Ctrl): n = 10, anti-CD49d: n = 13, P = 0.716. Data are shown as mean ± SD, linear-mixed-models, group-by-time interaction. Data were generated in two independent experiments. (C) The lesion volume was quantified at 7 and 28 d after stroke and shows no difference between treatment groups. Control 7 d: n = 6, control 28 d: n = 11, anti-CD49d 7 d: n = 5, anti-CD49d 28 d: n = 8, all WT animals. Data are shown as mean ± SD. Ordinary one-way ANOVA + Tukey’s post-hoc test. Data were generated in at least three independent experiments per time point. (D) Evaluation of the multi-parameter neuroscore and cylinder test shows no difference between treatment groups. Control: n = 10, anti-CD49d: n = 13, P (neuroscore) = 0.560, P (cylinder) = 0.691. Data are shown as median ± SE, linear-mixed models, group-by-time interaction. Data were generated in two independent experiments in Thy1-GCaMP6s animals. (E) Illustrative pixelmaps show the group-wise averaged seed-based FC between eight seeds representing functional cortical areas over an observation period of 84 d after stroke by longitudinal in vivo calcium imaging: 1, left rostral forelimb; 2, right rostral forelimb; 3, left caudal forelimb; 4, right caudal forelimb; 5, left forelimb sensory area; 6, right forelimb sensory area; 7, left hindlimb sensory area; 8, right hindlimb sensory area.. Gray squares indicate excluded seed autocorrelation, and white squares indicate excluded seeds due to the stroke lesion, when excluded in more than three animals. Data are shown as Fisher z-transformed connectivity scores and were generated in two independent experiments in Thy1-GCaMP6s animals. (F) Schematic illustration for longitudinal in vivo widefield calcium imaging acquisition; left intrahemispheric and whole cortex homotopic FC of the eight previously defined functional areas. Time course of left intrahemispheric FC (intra FC; P = 0.127) and overall homotopic FC (P = 0.295). Data are shown as median ± SE, linear-mixed models, group-by-time interaction. Data were generated in two independent experiments in Thy1-GCaMP6s animals. (G) Representative images of a Golgi-Cox–stained pyramidal neuron layer II/III in a naive animal (scale bar = 30 µm) and 3D reconstruction of pyramidal neuron dendrites (red) with dendritic spines (blue, scale bar = 5 µm). Dendritic spines were quantified in layer II/III in naive (untreated) animals and 84 d after stroke in stroke ipsi- and contralateral hemispheres. Naive: n = 5, control: n = 10, anti-CD49d: n = 13. Data are shown as median ± interquartile range. Wilcoxon rank-sum test with continuity correction and Bonferroni correction for multiple comparisons were used. Data were generated in at least two independent experiments. All data in this figure were obtained in a mixed-sex cohort (anti-CD49d: four male, nine female; control: three male, seven female), except for C (only male mice).