Fig. 6: Ccl2 deficiency reduces ocular and brain infiltration of monocytes/macrophages and BOB/BBB disruption in highly myopic mice.

a Flow-cytometric quantification showed decreased ratios of monocytes (CD11b⁺/CD45⁺), CD11b⁺/Ly6C⁺ monocytes, and macrophages (CD11b⁺/F4/80⁺) in the uvea of Ccl2-deficient highly myopic mice than in those of wild-type highly myopic mice. b Flow-cytometric quantification showed decreased ratios of CD11b⁺/Ly6C⁺ monocytes in the brain of Ccl2-deficient highly myopic mice than in the controls. c Immunofluorescent images showed decreased infiltration of Ly6C⁺ monocytes (arrows) in the BLA and ventral hippocampus of Ccl2-deficient highly myopic mice. Enlarged images in white squares are shown on the top right. Scale bars: 100 μm. d Representative images of enhanced MRI (coronal) showed slower leakage of contrast agent into the anterior chamber (yellow arrows) of Ccl2-deficient highly myopic mice than into those of wild-type highly myopic mice at both 12 min and 20 min after Gd-DTPA injection. e Immunofluorescent images of tight junction proteins occludin and ZO-1 in the ciliary body and retina of mice. Fluorescence intensity normalized to the averaged value of the control for quantification analyses. Scale bars: 25 μm. f Examination of occludin and ZO-1 expression in mouse retina with western blotting. The band density was normalized to that of the loading control for statistical analysis. The lane of ZO-1 for the wild-type highly myopic group was the same as that for the highly myopic group. g Leakage of EB into the mouse brain. h Relative fluorescence intensity of occludin in the BLA and ventral hippocampus area. Fluorescence intensity was normalized to the averaged value of the wild-type for quantification analyses. i Immunofluorescent images of occludin and CD31 in the BLA and ventral hippocampus of mice. Enlarged images of CD31-labeled vessels (square) showed upregulated occludin (arrowheads) in the Ccl2-deficient highly myopic mice. Scale bars: 40 μm in white and 20 μm in yellow. Data are mean ± SD. n = 6 in each group (a, b, g); n = 3 in each group (c–f, h). The level of significance was detected with one-way repeated measures ANOVA (d) or Student’s t-test (a, b, e–h). ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05.