Regulation of hippocampal H3 histone methylation by acute and chronic stress

Abstract

The hippocampal formation is a brain region noted for its plasticity in response to stressful events and adrenal steroid hormones. Recent work has shown that chromatin remodeling in various brain regions, including the hippocampus, is associated with the effects of stress in a variety of models. We chose to examine the effects of stress, stress duration, corticosterone administration, and fluoxetine treatment on the levels of hippocampal histone H3 methylation at lysines 4, 9, and 27, marks associated, respectively, with active transcription, heterochromatin formation, and transcriptional repression. We found that acute stress increased the levels of H3K9 tri-methylation (H3K9me3) in the dentate gyrus (DG) and CA1, while it reduced levels of H3K9 mono-methylation (H3K9me1) and H3K27 tri-methylation (H3K27me3) in the same regions, and had no effect on levels of H3K4 tri-methylation (H3K4me3). Seven days of restraint stress reduced levels of H3K4me3 in the CA1 and H3K27me3 in the DG and CA1, while increasing basal levels of H3K9me3. Chronic restraint stress (CRS) for 21 days mildly increased levels of H3K4me3 and reduced H3K9me3 levels in the DG. Treatment with fluoxetine during CRS reversed the decrease in DG H3K9me3, but had no effect on the other marks. These results show a complex, surprisingly rapid, and regionally specific pattern of chromatin remodeling within hippocampus produced by stress and anti-depressant treatment that may open an avenue of understanding the interplay of stress and hippocampal gene expression, and reveal the outlines of a potential chromatin stress response that may be diminished or degraded by chronic stress.

Fig. 1.

Levels of H3K4me3 immunoreactivity in hippocampus from rats either unstressed (basal), after acute stress, 24 h after 6 days of stress (7-day basal) or 7 days of restraint stress in the DG (A), CA1 (B), and CA3 (C).

Fig. 2.

Levels of H3K9me3 immunoreactivity in hippocampus from rats either unstressed (basal), after acute stress, 24 h after 6 days of stress (7-day basal) or 7 days of restraint stress in the DG (A), CA1 (B), and CA3 (C). (D) Representative sections through the hippocampus of unstressed (basal) and acutely restrained (ARS) rats. *, P < 0.05 vs. basal.

Fig. 3.

Levels of H3K9me1 immunoreactivity after acute stress in the DG (A) and CA1 (B). *, P < 0.05 vs. basal.

Fig. 4.

Levels of H3K27me3 immunoreactivity in hippocampus from rats either unstressed (basal), after acute stress, 24 h after 6 days of stress (7-day basal) or 7 days of restraint stress in the DG (A), CA1 (B), and CA3 (C). (D) Representative sections through the hippocampus of unstressed (basal) and acutely restrained (ARS) rats. *, P < 0.05 vs. basal.

Fig. 5.

Levels of H3K4me3 immunoreactivity in the DG after CRS or CRS with daily 10 mg/kg fluoxetine treatment. *, P < 0.05 vs. basal.

Fig. 6.

Levels of H3K9me3 immunoreactivity in the DG after CRS or CRS with daily 10 mg/kg fluoxetine treatment. *, P < 0.05 vs. basal.