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. 2022 May 6;8(18):eabm2545. doi: 10.1126/sciadv.abm2545

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Fig. 7. — (A) Strategy for the generation of the Rosa^Gpr56 mouse line. IRES, internal ribosomal entry site. (B) IHC staining showing reporter green fluorescent protein (GFP) is colocalized with Iba1 in the Rosa^Gpr56/+; Cx3cr1^Cre/+ mouse brain. (C) Reverse transcription qPCR (RT-qPCR) showing Gpr56 mRNA expression is largely increased in FACS-isolated microglia from Rosa^Gpr56/+; Cx3cr1^Cre/+ mice (n = 3). (D) Representative images of PV⁺ interneurons in the posterior SSC. (E) A bar graph showing PV⁺ interneuron density in the posterior SSC is decreased in global Gpr56 KO mice compared to control mice, and restoring microglial Gpr56 expression corrects the PV⁺ interneuron deficits. n = 8 (Cx3cr1^Cre/+), n = 6 (Gpr56 null; Cx3cr1^Cre/+), and n = 5 (Gpr56 null; Rosa^Gpr56/+; Cx3cr1^Cre/+). (F) Representative images of PV⁺ interneurons in the SSC of control mice (PBS, Cx3cr1^Cre/+), MIA offspring [poly(I:C), Cx3cr1^Cre/+], and MIA offspring with restored expression of microglial Gpr56 [poly(I:C), Rosa^Gpr56/+; Cx3cr1^Cre/+]. (G) A bar graph showing PV⁺ interneuron density is decreased in MIA offspring compared to controls. The density of PV⁺ interneurons is increased in MIA offspring with restored expression of microglial Gpr56 compared to control MIA offspring. n = 5 (PBS, Cx3cr1^Cre/+), n = 6 [poly(I:C), Cx3cr1^Cre/+], and n = 6 [poly(I:C), Rosa^Gpr56/+; Cx3cr1^Cre/+]. Unpaired t test for (C): **P < 0.01; two-way ANOVA and post hoc Bonferroni’s test for (E): ##P < 0.01 and ###P < 0.001 between Cx3cr1^Cre/+ and Gpr56 null; Cx3cr1^Cre/+; *P < 0.05, **P < 0.01, and ***P < 0.001 between Gpr56 null; Cx3cr1^Cre/+ and Gpr56 null; Rosa^Gpr56/+; Cx3cr1^Cre/+. Two-way ANOVA and post hoc Bonferroni’s test for (G): #P < 0.05 and ##P < 0.01 between PBS, Cx3cr1^Cre/+ and poly(I:C), and Cx3cr1^Cre/+; *P < 0.05 between poly(I:C), Cx3cr1^Cre/+ and poly(I:C), and Rosa^Gpr56/+; Cx3cr1^Cre/+. Data are presented as means ± SEM.