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. 2012 May 30;32(22):7499–7518. doi: 10.1523/JNEUROSCI.0392-12.2012

Figure 11.

Figure 11.

Immunohistochemical analyses on cerebellar slices of 45-d-old mice. A–D, According to the reporter DNA expression from two alleles, in cerebella of Cx30+/−/Cx47−/− and Cx30−/−/Cx47−/− mice, signals for eGFP are stronger compared to those in Cx30+/−/Cx47+/− and Cx30−/−/Cx47+/− mice, resulting in a higher background-to-signal ratio. In double-KO mice, the signals are further increased, which indicates higher activity of the Cx47 promoter in oligodendrocytes of double-deficient mice. In contrast, loss of eGFP-positive cells is evident in these mice compared to control littermates, especially in the granule cell layer (insets). E–H, MBP stainings revealed strong signals in all cerebella observed. Cx30−/−/Cx47−/− mice showed even increased MBP signals, indicated by the higher background-to-signal ratio of the staining. However, the immunosignals appeared condensed in the white matter of cerebellar lobes, and sponge like in the center of the cerebellum. Compared to Cx30+/−/Cx47+/− mice, Cx30+/−/Cx47−/− animals showed a mild reduction in size and number of myelin fibers pervading the granule cell layer in the cerebellum that was not detected in Cx30−/−/Cx47+/− mice (insets). A clear reduction in size and density of these myelin fibers was observed in cerebella obtained from Cx30−/−/Cx47−/− mice. I–L, GFAP-positive astrocytes were found in cerebellar slices obtained from all four genotypes with the typical parallel processes of GFAP-positive Bergmann glia fibers in the molecular layer. L, A conspicuous increase in GFAP signal intensity indicating astrogliosis was observed in the cerebellar white matter of Cx30−/−/Cx47−/− mice. Insets show higher magnifications of cerebellar white matter. M–P, Immunostainings revealed increased Iba1 expression of microglia in cerebella of Cx30−/−/Cx47−/− mice. In the white matter of Cx30+/−/Cx47−/− mice, some microglia also showed increased Iba1 expression, which was not found in cerebella obtained from Cx30+/−/Cx47+/− and Cx30−/−/Cx47+/− mice. Insets show high magnification of white matter tracts. Note that increased Iba1 expression is accompanied by morphological changes of these cells. Scale bars: A, E, I, M, 500 μm; insets, A, M, 50 μm; E, I, 200 μm.