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RNA sequencing microglia depletion model

ABSTRACT: We developed an experimental model system in mice, in which microglia can be depleted selectively and temporarily from the prefrontal cortex (PFC) during a defined window of adolescence (6 weeks of age). This was achieved using a single, bilateral stereotaxic intra-PFC injection of clodronate disodium salt (CDS), which induces selective apoptosis of microglia. To ascertain the cellular selectivity of intra-PFC administration of CDS, we conducted next-generation RNA sequencing (RNAseq) of bulk medial PFC tissue from CDS- or phosphate buffered saline (PBS)-injected mice. PFC samples were extracted either during the peak of microglia depletion, that is, 5 days post injection (5 dpi) or in adulthood (12 weeks of age) after full microglia repopulation.

ORGANISM(S): Mus musculus

PROVIDER: GSE168268 | GEO | 2022/03/01

REPOSITORIES: GEO

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Project description:Background: Epidemiological research indicates that iron deficiency (ID) in infancy correlates with long-term cognitive impairment and behavioral disturbances, despite therapy. However, the mechanisms underlying these effects are unknown. Objective: We investigated how ID affected postweaning behavior and monoamine concentration in rat brains to determine whether ID during the juvenile period affected gene expression and synapse formation in the prefrontal cortex (PFC) and nucleus accumbens (NAcc). Methods: Fischer344/Jcl male rats aged 21–39 days were fed low-iron diets (0.35 mg/kg iron; ID group) or standard AIN-93 G diets (3.5 mg/kg iron; control group). The locomotor activity of male offspring was evaluated by the open field and elevated plus maze tests at ages 8 and 12 weeks. Monoamine concentrations in the PFC, NAcc, caudate-putamen, ventral midbrain, dorsal midbrain, and pons were analyzed. Comprehensive gene expression analysis was performed in the PFC and NAcc at age 13 weeks. Finally, we investigated synaptic density in the PFC and NAcc by synaptophysin immunostaining. Results: Behavioral tests revealed significant interactions between age and iron consumption for the total distance traveled and the distance traveled in the peripheral area (p < 0.05), indicating that ID during the juvenile period affected hyperactivity and that this persisted to adulthood. At age 13 weeks, the ID group had increased levels of both dopamine and the metabolites of dopamine and serotonin in the NAcc. Comprehensive gene expression analysis and immunostaining showed decreased Reelin gene expression (adjusted p < 0.01) and significantly increased spine density in the NAcc in the ID group compared with the control group (p < 0.01). Conclusions: ID during the postweaning juvenile period led to long-term hyperactivity, monoamine disturbance in the brain, and downregulation of Reelin expression in the NAcc despite complete iron repletion. Epigenetic modification of Reelin genes may be involved in synaptic plasticity in the NAcc.

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RNA sequencing microglia depletion model

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