内侧前额叶皮质DNA甲基化调控大鼠酒精相关行为

doi:10.16288/j.yczz.19-261

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Hereditas(Beijing) ›› 2020, Vol. 42 ›› Issue (1): 112-125.doi: 10.16288/j.yczz.19-261

• Research Article • Previous Articles Next Articles

DNA methylation in the medial prefrontal cortex regulates alcohol-related behavior in rats

Hengzhen Cui¹, Mizhu Sun², Runzhi Wang², Chenyu Li³, Yuxuan Huang³, Qiuju Huang³, Xiaomeng Qiao²()

^1. Basic Medicine, School of Medicine, Zhengzhou University, Zhengzhou 450001, China
^2. Department of Forensic Medicine, School of Medicine, Zhengzhou University, Zhengzhou 450001, China
^3. Department of Medical Laboratory,School of Medicine, Zhengzhou University, Zhengzhou 450001, China

Received:2019-09-02 Revised:2019-11-21 Online:2020-01-20 Published:2019-12-25
Contact: Qiao Xiaomeng E-mail:xiaomeng416520@126.com
Supported by:
Supported by the National Natural Science Foundation of China No(81373247);the Innovative Project for College Students of Zhengzhou University No(2019cxcy054)

Abstract

Abstract:

Alcohol abuse causes tissue and organ damage, and may participate neuropsychiatric diseases. Studies have shown that DNA methylation plays an important role in gene expression and behavioral changes induced by alcohol, however the causative neurobiological mechanisms have not been clarified. In this study, 32 healthy adult male SD rats were randomly divided into a drinking water control group (n=16) and a chronic alcohol exposure group (n=16). The alcohol preference and locomotor activity of rats were evaluated by two-bottle choice test (TBCT) and open-field test (OFT). DNA methylation in the medial prefrontal cortex (mPFC) tissue was detected by the reduced representative bisulfite sequencing (RRBS) technology. The methylation differential genes closely related to alcohol abuse were screened. qRT-PCR was used to verify the mRNA expression patterns of differential genes. qRT-PCR and Western blot were used to detect the expression of DNA methyltransferases (DNMTs) and methyl CpG binding protein 2 (MeCP2). Furthermore, the effect of short-term alcohol exposure (7 days) on DNMTs and MeCP2 in the mPFC of rats was tested (n=8/group). The results indicated that the methylation level of promoter region in the mPFC of rats exposed to chronic alcohol was significantly increased. In addition, the increased methylation levels in the promoter of Ntf3 and Ppm1G were accompanied by down-regulated mRNA levels in the chronic alcohol exposure group. The decreased methylation levels in the promoter of Hap1 and DUSP1 were accompanied by up-regulated mRNA levels. Furthermore, chronic alcohol exposure increased the mRNA and protein levels of DNMT3B and MeCP2. However, short term alcohol exposure did not affect their expression. This present study provides evidence that DNA methylation is associated with the development of alcohol abuse, which may be regulated by DNMT3B and MeCP2. The target genes Ntf3, Ppm1G, Hap1, and DUSP1 related to alcohol abuse were discovered as well, providing new insights into the neurobiological mechanism of alcohol abuse and the potential pharmacological targets for the treatment of alcohol abuse.

Key words: alcohol abuse, DNA methylation, DNA methyltransferases, medial prefrontal cortex

Hengzhen Cui, Mizhu Sun, Runzhi Wang, Chenyu Li, Yuxuan Huang, Qiuju Huang, Xiaomeng Qiao. DNA methylation in the medial prefrontal cortex regulates alcohol-related behavior in rats[J]. Hereditas(Beijing), 2020, 42(1): 112-125.

Figures/Tables 7

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基因名称	引物序列(5′→3′)
Ntf3	F: GGCAACAGAGACGCACAAT
Ntf3	R: TCCTCCGTGGTGATGTTCTA
Pdp1	F: CTCAATTACCTGGTGCTTCG
Pdp1	R: GCATTGTGGTCATTAGAGAGC
Ppm1G	F: TCATTGCTTGTGACGGCATC
Ppm1G	R: ATGACAATAACCGAAGCTCCC
FLT1	F: TAACGCAATCTTCTTAACGTC
FLT1	R: TTAATCCTCACAAACGTCCC
N1g2	F: CAGAAGAGACAGAGCAGGCATC
N1g2	R: GCAGAGACCACCACATTCATC
Hap1	F: CAAGGATGAATGTGGTGGTCTC
Hap1	R: CTGCTGCCGCTTAGAATGG
NFκB-p100	F: TCGACCTCCACCGGATCTTT
NFκB-p100	R: CTGTGTCTTTTGGCTTCGGC
RTKN2	F: CCATAGGATGCTAATCTCCGTTT
RTKN2	R: AAGATTACTGCGGTCAACCTT
DUSP1	F: CATTCCCGATGACATACACGTT
DUSP1	R: CATTCCCGATGACATACACGTT
DUSP8	F: TCACTGGTCTGGCTGGCATTTC
DUSP8	R: AGTTGAAGTTGGGCGAGATGG
DNMT1	F: GTTCCTTGTAGGCGAGTGTG
DNMT1	R: TTGCGTAGTCCTGGCTGTAC
DNMT3A	F: GCA AAGTGAGGACCATTACC
DNMT3A	R: GCCAAACACCCTTTCCAT
DNMT3B	F: GTGCGTCGTTCAGGCAGTAG
DNMT3B	R: GTTCTCGGCTCTCCTCATCT
MeCP2	F: GCCTTCAGCCCACCATTCTG
MeCP2	R: CGGTCACGAATGATGGAACG
GAPDH	F: ATGGGGAAGGTGAAGGTCG
GAPDH	R: GGGGTCATTGATGGCAACAA

DNA methylation in the medial prefrontal cortex regulates alcohol-related behavior in rats

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