Dynamic methylation driven by neuronal activity in hippocampal neurons impacts complex behavior

doi:10.1007/s11515-015-1369-8

Front. Biol.

2015, Vol. 10

Issue (5) : 439-447 https://doi.org/10.1007/s11515-015-1369-8

RESEARCH ARTICLE

Dynamic methylation driven by neuronal activity in hippocampal neurons impacts complex behavior

Anita E. Autry,Megumi Adachi,Lisa M. Monteggia(

)

Department of Neuroscience, The University of Texas Southwestern Medical Center, Dallas, TX 75390-9070, USA

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Abstract

Epigenetic processes are well-known to play critical roles in learning and memory. Among epigenetic processes, accumulating data suggests that DNA methylation in particular is a critical determinant of learning and memory. In vitro data have suggested that DNA methyltransferase inhibitors can trigger DNA demethylation and subsequent gene expression of the brain-derived neurotrophic factor gene in an activity dependent manner. To examine if these processes occur in vivo, we chronically infused DNMT inhibitors into the hippocampus and examined the impact on behavior. We find that chronic DNMT inhibition in the hippocampus results in increased anxiety-related behavior and deficits in context-dependent fear conditioning accompanied by an increase in BDNF expression. Gene expression changes were blocked by pretreatment with the NMDA receptor antagonist AP5, suggesting that DNMT inhibition enhances gene expression in an activity-dependent manner and that, conversely, the behavior deficits and abnormal gene expression are facilitated by NMDA receptor activity.

Keywords DNA methylation NMDA receptors learning and memory behavior hippocampus

Corresponding Author(s): Lisa M. Monteggia

Just Accepted Date: 17 August 2015 Online First Date: 17 September 2015 Issue Date: 30 October 2015

Cite this article:

Anita E. Autry,Megumi Adachi,Lisa M. Monteggia. Dynamic methylation driven by neuronal activity in hippocampal neurons impacts complex behavior[J]. Front. Biol., 2015, 10(5): 439-447.

URL:

https://academic.hep.com.cn/fib/EN/10.1007/s11515-015-1369-8
https://academic.hep.com.cn/fib/EN/Y2015/V10/I5/439

Fig.1 Overview of the infusion paradigm. (A) Mice were implanted with bilateral cannulae directed toward the dentate gyrus attached to a minipump and allowed to recover from surgery for six days. Starting on day 7, mice were tested for behavioral impact of chronic infusion of either vehicle or DNMT inhibitors 5azaC or zebularine (Zeb) including elevated plus maze, social interaction, locomotion, and fear conditioning. (B) Cannula placement was confirmed by examining the location of the cannula track in cressyl violet stained tissue.

Fig.2 Chronic DNMT inhibitor infusion enhances anxiety-like behavior. (A, C) Mice treated with DNMT inhibitors 5azaC (A) or zebularine (C) spend less time exploring the open arm of the elevated plus maze compared with vehicle-infused animals (*p<0.05). (B, D) Similarly, mice treated with 5azaC (B) or zebularine (D) spend more time in the closed arm of the EPM compared to vehicle-treated mice (*p<0.05).

Fig.3 Chronic DNMT inhibitor treatment does not impact locomotor activity. Treatment by 5azaC (A) or zebularine (B) does not alter locomotor behavior .

Fig.4 Chronic inhibition of DNMT activity in the dentate gyrus impairs context dependent fear memory. (A,B). Mice receiving (A) 5azaC or (B) zebularine spent significantly less time freezing in the context in which fear training was administered the previous day compared to vehicle-treated mice (*p<0.05).

Fig.5 Blockade of excitatory signaling blocks effects of DNMT inhibition. (A) Chronic DNMT inhibition results in increased anxiety-related behavior (F_3-37 = 2.852, p = 0.05, main effect by one-way ANOVA) and this effect is blocked by co-treatment with AP5 (significant post-hoc analysis after Neuman-Keul’s correction, 5azaC vs. AP5+ 5azaC groups). (B) As previously shown, DNMT inhibition impairs context-dependent fear memory (F₃₋₃₇ = 2.759, p = 0.05, main effect by one-way ANOVA, significant post-hoc analysis Neuman-Keul’s correction, vehicle vs. 5azaC) while co-treatment of 5azaC+ AP5 is indistinguishable from vehicle treatment, though AP5 alone produces no effect on contextual fear memory. (C) Quantitative reverse transcription PCR reveals that DNMT inhibition does not appear to globally enhance transcription, but specifically regulates Bdnf levels in the dentate gyrus (F₃₋₂₃ = 3.263, p = 0.04) (post-hoc analysis by Dunnett’s test between vehicle and 5azaC, *p<0.05). This effect is blocked by antagonism of NMDA receptors by AP5, while AP5 treatment alone does not affect bdnf or other transcript levels (F₃₋₂₃ = 0.3418, p = 0.79 for reelin; F₃₋₂₃ = 0.6404, p = 0.59 for PP1, one-way ANOVA).

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