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Inhibition of microRNA-34a Suppresses Epileptiform Discharges Through Regulating Notch Signaling and Apoptosis in Cultured Hippocampal Neurons

  • Jinli Wang
  • Yuan Zheng
  • Xu Cheng
  • Fenfen Xu
  • Piaopiao Zhang
  • Xiao Zhou
  • Hongyang ZhaoEmail author
Original Paper
  • 16 Downloads

Abstract

Epilepsy is characterized by recurrent unprovoked seizures and some seizures can cause neuronal apoptosis, which is possible to make contributions to the epilepsy phenotype, impairments in cognitive function or even epileptogenesis. Moreover, many studies have indicated that microRNA-34a (miRNA-34a) is involved in apoptosis through regulating Notch signaling. However, whether miRNA-34a participates in neuronal apoptosis after seizures remain unclear. Therefore, we aimed to explore the expression of miRNA-34a and its effects on the epileptiform discharge in spontaneous recurrent epileptiform discharges (SREDs) rat hippocampal neuronal pattern. Mg2+-free medium was used to induce SREDs, quantitative reverse-transcription polymerase chain reaction was used to detect the expression of miRNA-34a, western blot was used to determine the expression of Notch pathway and apoptosis-related proteins, and whole cell current clamp recordings was used to observe the alteration of epileptiform discharge. We found obvious apoptosis, increased expression of miRNA-34a and decreased expression of Notch signaling in Mg2+-free-treated neurons. Treatment with miRNA-34a inhibitor decreased the frequency of action potentials, activated Notch signaling and prevented neuronal apoptosis in Mg2+-free-treated neurons. However, treatment with miRNA-34a mimics increased the frequency of action potentials, down-regulated Notch signaling and promoted neuronal apoptosis in Mg2+-free-treated neurons. Furthermore, γ-secretase inhibitor N-[N-(3,5-di-uorophenacetyl)-1-alanyl]-S-phenylglycine t-butylester (DAPT), an inhibitor of Notch signaling, could weaken anti-apoptosis effect of miRNA-34a inhibitor. These results suggest that inhibition of miRNA-34a could suppress epileptiform discharges through regulating Notch signaling and apoptosis in the rat hippocampal neuronal model of SREDs.

Keywords

Epilepsy MicroRNA-34a Notch signaling Apoptosis 

Abbreviations

SREDs

Spontaneous recurrent epileptiform discharges

qRT-PCR

Quantitative reverse-transcription polymerase chain reaction

MAP2

Protein-2 associated with anti-microtubule

pBRS

Physiological basal recording solution

AP

Action potentials

Bax

Bcl-2-associated X protein

Bcl-2

B-cell lymphoma-2

HRP

Horseradish peroxidase

NIC

Notch intracellular domain

Notes

Compliance with Ethical Standards

Conflict of interest

The authors declare that there are no conflicts of interest.

Ethical Approval

Every animal use process was supported by the Animal Care and Use Committee of Shandong University and was in severe accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Jinli Wang
    • 1
  • Yuan Zheng
    • 1
  • Xu Cheng
    • 2
  • Fenfen Xu
    • 1
  • Piaopiao Zhang
    • 2
  • Xiao Zhou
    • 1
  • Hongyang Zhao
    • 1
    Email author
  1. 1.Department of PediatricsJinan Central Hospital, Shandong UniversityJinanChina
  2. 2.Clinical Medical CollegeTaishan Medical UniversityTaianChina

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