Gene expression analysis of mouse BNST and blood cells after chronic social defeat stress (CSDS)
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Chronic psychosocial stress is a risk factor for psychiatric disorders, and genetic factors interact in conferring susceptibility. The chronic social defeat stress (CSDS) mouse model allows identifying factors underlying resilience and susceptibility to chronic psychosocial stress. Following 10 days of CSDS, we collected the brain and blood tissues. We performed (1) AGO2 RNA immunoprecipitation-sequencing (AGO2 RIP-seq) of active miRNAs and their mRNA targets in the BNST. Additionally, we carried out RNA-seq and miRNA-seq from blood cells. We performed ribo-zero globin depletion of RNA from blood cells. Immunoprecipitation of AGO2-bound miRNAs and their target mRNAs was done by pooling of 3 BNST collected from all the phenotypic groups in both strains. We sequenced with Illumina Next-seq 500 from (1) BNST AGO2 mRNA (C57BL/6Crl: control n = 5, resilient n = 5, susceptible n = 4), (2) BNST AGO2 miRNA (C57BL/6Crl: control n = 5 , resilient n =5 , susceptible n = 3), (3) blood cells mRNA (C57BL/6Crl: control n = 5, resilient n = 5, susceptible n = 5; DBA/2Crl: control n = 5, susceptible n = 5), (4) blood cells miRNA (C57BL/6Crl: control n = 5, resilient n = 5, susceptible n = 5; DBA/2Crl: control n = 5, susceptible n = 5) 6-8 days after the last CSDS.
长期社会心理应激是精神障碍的危险因素，且遗传因素可与之交互，赋予机体疾病易感性。慢性社会挫败应激（chronic social defeat stress, CSDS）小鼠模型可用于解析慢性社会心理应激的抵抗性与易感性相关的潜在调控因子。在为期10天的CSDS造模结束后，我们采集了脑组织与血液组织。 我们开展了以下实验：(1) 对终纹床核（bed nucleus of the stria terminalis, BNST）中活性微小RNA（microRNA, miRNA）及其mRNA靶标进行AGO2 RNA免疫共沉淀测序（AGO2 RNA immunoprecipitation-sequencing, AGO2 RIP-seq）；此外，我们还对血细胞进行了RNA测序（RNA-seq）与miRNA测序，并利用ribo-zero对血细胞RNA进行珠蛋白降解处理。 为完成AGO2结合miRNA及其靶mRNA的免疫共沉淀实验，我们将两个品系中所有表型组来源的3个终纹床核样本进行混合。我们于末次CSDS造模后6~8天，使用Illumina Next-seq 500平台对以下样本进行测序： (1) 终纹床核AGO2结合mRNA样本（C57BL/6Crl品系：对照组n=5、抵抗组n=5、易感组n=4）； (2) 终纹床核AGO2结合miRNA样本（C57BL/6Crl品系：对照组n=5、抵抗组n=5、易感组n=3）； (3) 血细胞mRNA样本（C57BL/6Crl品系：对照组n=5、抵抗组n=5、易感组n=5；DBA/2Crl品系：对照组n=5、易感组n=5）； (4) 血细胞miRNA样本（C57BL/6Crl品系：对照组n=5、抵抗组n=5、易感组n=5；DBA/2Crl品系：对照组n=5、易感组n=5）。
