Identification of the transcription factor, AFF4, as a new target of miR-203 in CNS

Linpeng Li,Yi Zou,Shufang Li,Jia Gan,Yaohui Liang,Lin Cao,Xiaosheng Liang

doi:10.1016/j.ijbiomac.2021.04.089

Linpeng Li, Yi Zou + Show 5 more

Open Access

https://doi.org/10.1016/j.ijbiomac.2021.04.089

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Abstract

MiR-203 was identified as a hub of a potential regulatory miRNA network in central nervous system. Overexpressing of miR-203 in the frontal cortex of C57BL/6J wild type mouse induced neurodegeneration by increasing the apoptotic pathway and neuron death. AFF4, a transcription factor, was identified as a new bona fida protein target of miR-203 in CNS. The miRNA:mRNA interaction of miR-203 and AFF4 was verified using Dural-luciferase assay. Down-regulated expression of AFF4 was induced by overexpressing miR-203 both in vitro and in vivo. Open field test, Y maze and Morris water maze test were conducted for the behavioral assessment of the mice with stereotactic injection of lentiviral vector overexpressing miR-203 in the hippocampus. No anxiety-like behavior or impaired cognition was noticed in these mice. Consistent with the results of the behavioral assessment, the electron micrograph and Nissl staining revealed no significant change in the synaptic density and no neuron injuries in the hippocampus of mice overexpressing miR-203, respectively. Our results indicated that instead of promoting neurodegenerative phenotype, a more profound function should be ascribed to miR-203 in regulating neuron behavioral activities and cognition. Neuron-type specific functions of miR-203 are likely to be executed via its various downstream protein interactors.

Highlights

Volatile anesthetics displayed profound influence on perioperative neurocognition, with controversial evidence suggesting both neurotoxicity and neuroprotection [1]
Given that miR-203 was found to target the proteins with opposite regulatory functions, our results indicated that instead of promoting neurodegenerative phenotype, a more profound function should be ascribed to miR-203 in regulating neuron behavioral activities and cognition
Dulbecco's modified Eagles medium (DMEM) and fetal bovine serum (FBS) used in cell culture were purchased from Invitrogen. 3,3diaminobenzidine tetrachloride (DAB), 1,4-dithiothreitol (DTT), Dimethyl Sulfoxide (DMSO), Triton X-100, Paraformaldehyde, Bovine Serum Albumin, Ethanol, and KCl hematoxylin protease inhibitor cocktail, Penicillin and streptomycin were from Sigma

Summary

Introduction

Volatile anesthetics displayed profound influence on perioperative neurocognition, with controversial evidence suggesting both neurotoxicity and neuroprotection [1]. We observed that overexpressing miRNA-203 in vitro significantly increased the tolerance of cultured B35 cells to OGD, possibly via promoting Akt phosphorylation and in turn, improving cell surviving, indicating a potential role of neuroprotection of miRNA-203. The expression of endogenous miR-203 was found abundant in keratinocytes and could be further elevated under chronic inflammatory skin conditions Both pro-inflammatory cytokines, TNFa and IL24, and the suppressor of cytokine signaling (SOCS3 and SOCS6) were identified as direct target of miR-203 [10]. These findings suggested a role of miR-203 in fine-tuning the immune response in these cells. Instead of promoting neurodegenerative phenotype, our results suggested a more profound function of miR-203 in regulating neuron behavioral activities and cognition

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