Dynamic Change of Shanks Gene mRNA Expression and DNA Methylation in Epileptic Rat Model and Human Patients

Abstract

Synaptic protein shanks (SH3 and multiple ankyrin repeat domains protein, Shank) have emerged as an important mediator of synaptic remodeling. Synaptic remodeling is a common pathogenic process in various neurological disorders including epilepsy. However, the expression and function of shanks gene in epileptogenesis has not been investigated to date. Herein, we investigated the expression of shanks (shank1/2/3) mRNA expression in both epileptic rats and epilepsy patients. Furthermore, methyl target sequencing was applied to explore the relationship between shank mRNA expression and DNA methylation in both rats and human patients. In general rat model, shank1/2/3 mRNA was downregulated at acute stage, upregulated at latent stage, and returned to the basal level at chronic stage. Ten CpG sites of shank1 was found differentially methylated, out of which 6 were hypermethylated. Seventeen CpG sites of shank3 were differentially methylated, out of which 8 were hypermethylated. In human epilepsy patients, decreased shank2 mRNA was detected from the brain tissue, with DNA hypermethylation dominant from both brain (18 out of 30) and blood tissue (58 out of 80), indicating the regulation role of DNA methylation on shank2 expression. In conclusion, our finding suggests the participation of the shanks gene in the pathophysiology of seizure, out of which 2 shank3 CpG sites (Chr7: 130473419, and Chr7: 130473405) may play an important role in shank3 expression at both the acute and latent stages in the SE rat model.

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Funding

This work was supported through funding from the National Natural Science Foundation of China (grant numbers 81671299 to B.X., and 81701182 to Y.F.) and the Natural Science Foundation of Hunan Province (grant number: 2018JJ2648) to Z.L.

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Authors

Contributions

Du Liu and Zhaohui Luo conducted the majority of the experiments. Yujiao Fu analyzed and interpreted the data and drafted the manuscript. Jialing Guo and Wei Xiao prepared the epilepsy animal experiment. Du Liu and Zhaohui Luo prepared the CpG detective experiments. Hongyu Long and Wenbiao Xiao analyzed and interpreted the data. Li Feng prepared for language editing and revising. Bo Xiao and Zhaohui Luo contributed to the study design. Bo Xiao provided financial support. All authors consent to the publication of the manuscript.

Corresponding authors

Correspondence to Zhaohui Luo or Bo Xiao.

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The authors declare that they have no conflict of interest.

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All procedures were followed in accordance with the ethical standards of the responsible committee on animal and human experimentation (the Ethics Review Committee of Xiangya Hospital, numbers 201603296 and 201603297) and with the Helsinki Declaration of 1964 and later versions.

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Electronic Supplementary Material

ESM 1

(DOC 79 kb)

ESM 2

The CpG distribution of shank1 in the human brain tissue and the rat brain. (PNG 470 kb)

ESM 3

The CpG distribution of shank2 in the human brain tissue and the rat brain. (PNG 483 kb)

ESM 4

The CpG distribution of shank3 in the human brain tissue and the rat brain. (PNG 523 kb)

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Fu, Y., Liu, D., Guo, J. et al. Dynamic Change of Shanks Gene mRNA Expression and DNA Methylation in Epileptic Rat Model and Human Patients. Mol Neurobiol 57, 3712–3726 (2020). https://doi.org/10.1007/s12035-020-01968-5

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Keywords

  • Shanks gene
  • Epilepsy
  • Methylation
  • CpG
  • Synapse
  • Human patients