Microglial transcriptome of Maternal Immune Activation model at different developmental Stages and after microglia repopulation. Microglial transcriptome of Maternal Immune Activation model at different developmental Stages and after microglia repopulation

To determine the transcriptomic changes underlying the MIA microglia phenotype and their reversal by PLX, we performed RNA-sequencing of magnetic CD11B beads-isolated microglia from the whole brain in Saline and MIA male and female offspring at E17, P7, P20, and P60, as well as from MG-REP male and female offspring at P60. Microglial transcriptome reveals novel MIA and repopulation modules and overlap of MIA microglial genes with ASD gene network. 14,225 microglial genes from RNA-seq data revealed distinct transcriptional signatures of immature microglia (IM module, 4681 transcripts, enriched in E17 and P7 Saline), MIA immature microglia (MIA-IM module, 2816 transcripts, enriched in E17 and P7 MIA), Juvenile microglia (JM module, 2318 transcripts, enriched in P20 Saline and MIA), Adult microglia (AM module, 3276 transcripts, enriched in P60 Saline + CTRL, P60 MIA + CTRL and P60 MIA + MG-REP), and repopulated adult microglia (REP-AM module, 1,134 transcripts, enriched in P60 Saline + MG-REP) Overall design: CD11b+ Microglia mRNA transcriptome profiling using RNA-seq Illumina HiSeq2000 across different development periods consisting of E17, P7, P20, and P60 from saline and MIA treated mothers. At P60, mice from saline and MIA treated mothers were treated with control or CSF1R inhibitor PLX5622 (MG-REP group)为明确母体免疫激活（Maternal Immune Activation, MIA）小胶质细胞表型背后的转录组学变化及其被PLX（PLX5622）逆转的机制，我们对生理盐水组与MIA模型组雌雄仔鼠在E17、P7、P20及P60时，全脑经磁珠分离得到的CD11B阳性小胶质细胞开展了RNA测序（RNA-seq）；同时纳入P60时MG-REP组雌雄仔鼠的样本。小胶质细胞转录组分析揭示了全新的MIA相关及小胶质细胞重填充模块，且MIA小胶质细胞基因与自闭症谱系障碍（Autism Spectrum Disorder, ASD）基因网络存在重叠。我们从RNA-seq数据中鉴定出14225个小胶质细胞基因，可分为具有独特转录特征的多个模块：未成熟小胶质细胞模块（IM模块，含4681个转录本，富集于E17和P7的生理盐水组）、MIA相关未成熟小胶质细胞模块（MIA-IM模块，含2816个转录本，富集于E17和P7的MIA模型组）、幼年小胶质细胞模块（JM模块，含2318个转录本，富集于P20的生理盐水组与MIA模型组）、成年小胶质细胞模块（AM模块，含3276个转录本，富集于P60生理盐水+对照组、P60 MIA+对照组及P60 MIA+MG-REP组），以及重填充成年小胶质细胞模块（REP-AM模块，含1134个转录本，富集于P60生理盐水+MG-REP组）。实验整体设计：采用基于Illumina HiSeq2000平台的RNA-seq技术，对经生理盐水或MIA处理母鼠所产仔鼠在E17、P7、P20及P60不同发育阶段的CD11B阳性小胶质细胞的mRNA转录组进行分析；在仔鼠P60时，对经生理盐水或MIA处理母鼠所产的仔鼠分别施加对照或集落刺激因子1受体（Colony Stimulating Factor 1 Receptor, CSF1R）抑制剂PLX5622处理，对应MG-REP组。