Fig. 6

CCL3 expression upregulation in microglia during development induced by prenatal VPA exposure. VPA (800 mg/kg) was orally administered on E11.5. Minocycline (100 mg/kg) was administered to dams by drinking water from P1 to P10. A Total RNA was extracted from the hippocampus, and CCL3 mRNA levels were determined by qPCR. The values are presented as the mean ± S.D. (n = 4 male pups from 2 dams in each group). The data were analyzed using two-way ANOVA [F(1, 12) = 11.82, p = 0.0049] followed by Tukey's t test. B CCL3 protein expression in the hippocampus was measured by ELISA. The values are presented as the mean ± S.D. (n = 8 male pups from 2 dams in each group). The data were analyzed using two-way ANOVA [F(1, 28) = 15.46, p = 0.0005] followed by Tukey's t test. C, D CD11b-positive cells were isolated from the hippocampi of male P10 mice, and then CCL3 levels were measured by western blotting. Representative images are shown in panel C, and the results of quantitative analysis of the bands are presented in panel D. Hippocampi collected from 5 male pups were used to isolate CD11b-positive cells. Three independent experiments were performed with 5 litters (15 male vehicle pups from 3 dams and 15 male VPA pups from 3 dams were used for this experiment). The values are presented as the mean ± S.D. The data were analyzed using Student's t test. E CCL3 mRNA expression in the cortex and amygdala in P10 mice was evaluated by qPCR. F CCR1 and CCR5 mRNA levels in the hippocampi of P10 mice were measured. G CCL3 mRNA expression was measured in the P1 mouse brain. The values are presented as the mean ± S.D. The data were analyzed using Student's t test