Ccn1 deletion in neural stem progenitor cells (NSPCs) leads to a reduction of microglia in the brain at E17.5 and P2 (IMAGE)
Caption
(A) Immunohistochemistry analysis of CCN1 in wild type (WT) and conditional knockout (CKO) brains at E13.5, E16.5, and P0. Scale bar: 50 μm. (B) Expression of Ccn1 in NSPCs of the mouse brain at E16.5, P0, and P2 from published data. (C) CCN1 is highly expressed in NSCs at E17.5 and P2, as confirmed by co-immunostaining of CCN1 (green) and SOX2 (silver). Scale bar: 50 μm. (D) Schematic diagram of the generation of Ccn1-CKO mice using Nestin-Cre. (E, F) Confirmed the expression specificity and CKO efficiency of Ccn1 in WT and CKO mouse brains at E17.5 (E) and P2 (F), as validated by scRNA-seq.(G) CCN1 expression is nearly absent in E17.5 and P2 CKO mouse brains, as shown by co-immunostaining of DAPI (blue) and CCN1 (red). Scale bar: 50 μm. (H) The percentage of microglia was significantly reduced at E17.5 and P2 according to scRNA-seq analysis. (I) The proportion of microglia in the brain was significantly decreased upon Ccn1 depletion in NSPCs at E17.5 and P2, as determined by FACS analysis using *tmem119-tdTomato; Nestin-cre; Ccn1flox/flox* mice (n = 6 brains per group; E17.5, P = 0.0071; P2, P < 0.0001). (J, K) Microglia were markedly reduced in the ventricular zone (VZ), as shown by immunofluorescence of the microglial marker IBA1 (J) and statistical analysis of IBA1+ nuclei in the VZ (K) (n = 6 mice per group; E17.5, P < 0.001; P2, P < 0.0001). Scale bar: 50 μm. Data information: In (I) and (K), data are presented as mean ± SEM. Unpaired Student’s t-test (two-tailed) was used: ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001.
Credit
Zhile Bai, Zhanhe Chang, Shuguang Wang, Jiangli Zheng, Huan Wu, Hanyu Liu, Yalin Zhang, Jiayang Xie, Qingran Bai, Jiqing Yin, You Wu, Qin Shen, Shaorong Gao, Yawei Gao
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