Effects of Minocycline and EE in CUMS as a MDD model: transcriptomic profiles and synaptic plasticity of microglia

Two independent differential expression analyses were performed in mice subjected to chronic unpredictable mild stress (CUMS): one consisted of microglia (Cd11b+ cells) from the prefrontal cortex (PFC) and the other from the hippocampus (HC). After the CUMS protocol, minocycline, a wide spectrum antibiotic with anti-inflammatory properties, was delivered (CUMS_M group) in the drinking water/saccharine solution during 21 days. Transcriptomic profiling for a vehicle group (VEH), who spent 3 weeks in the cage with the same drinking solutions (Water/ saccharine 0.1%) and for a environmental enrichment (EE) group, that also spent 3 more weeks in the cage where enrichment materials were provided, were performed as well. Overall design: All bulk mRNA expression profiles were acquired from isolated PFC and HC areas. CD11b+ cells were sorted using the purity precision mode on FACS Aria. RNA was isolated following the protocol described in RNeasy kit manual and RNA concentration was immediately determined using a spectrophotometer (NanoDrop2000 ThermoScientific). Quality and integrity of total RNA was assessed with Agilent 2100 Bioanalyzer and strand-specific polyA enriched RNA libraries were prepared using the KAPA Stranded mRNA Sample Preparation Kit. Then, libraries were run in the rapid run flow cell -paired-end sequenced (2x76bp) - on HiSeq 1500. RNA sequencing analysis revealed several differentially expressed genes between groups in both PFC and HC.