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Direct Current Electric Field Coordinates the Migration of BV2 Microglia via ERK/GSK3β/Cofilin Signaling Pathway

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Abstract

Direct current electric field (DCEF) steers the migration of various neural cells. Microglia, as macrophage of the central nervous system (CNS), however, have not been reported to engage in electrotaxis. Here, we applied electric fields to an in vitro environment and found directional migration of BV2 microglia toward the cathode, in a DCEF strength-dependent manner. Transcriptome analysis then revealed significant changes in the mitogen-activated protein kinase cascades. In terms of mechanism, DCEF coordinated microglia movement by regulating the ERK/GSK3β/cofilin signaling pathway, and PMA (protein kinase C activator) reversed cell migration through intervention of the ERK/GSK3β/cofilin axis. Meanwhile, LiCl (GSK3β inhibitor) showed similar functions to PMA in the electrotaxis of microglia. Furthermore, pharmacological and genetic suppression of GSK3β or cofilin also modulated microglia directional migration under DCEF. Collectively, we discovered the electrotaxis of BV2 microglia and the essential role of the ERK/GSK3β/cofilin axis in regulating cell migration via modulation of F-actin redistribution. This research highlights new insight toward mediating BV2 directional migration and provides potential direction for novel therapeutic strategies of CNS diseases.

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Data Availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

TEPs:

Transepithelial electrical potentials

ADF:

Actin depolymerizing factor

DCEF:

Direct current electric field

CNS:

Central nervous system

NGS:

Next-generation RNA sequencing

hNSCs:

Human neural stem cells

MAPK:

Mitogen activated protein kinases

ERK:

Extracellular signal-regulated kinase

GSK3β:

Glycogen synthase kinase-3β

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

PMA:

Phorbol-12-myristate-13-acetate

DMSO:

Dimethyl sulfoxide

cDNA:

Complementary DNA

cRNA:

Complementary RNA

KEGG:

Kyoto encyclopedia of genes and genomes

GSEA:

Gene set enrichment analysis

DEPC:

Diethylpyrocarbonate

PBS:

Phosphate-buffered saline

RT:

Room temperature

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Acknowledgements

We would like to thank Professor Jiawei Zhou from the Shanghai Institutes for Biological Science, Chinese Academy of Sciences (Shanghai, China) for providing us with BV2 cells.

Funding

This work was sponsored by the National Natural Science Foundation of China (grant numbers 82071358, 81771317, 81772058 and 81801235), Shanghai Talent Development Fund (grant number 2018077), Program of Shanghai Academic/Technology Research Leader (grant number 21XD1422400) and Project of Shanghai Medical And Health Development Foundation (20224Z0012).

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Author notes

  1. Yuxiao Ma, Chun Yang, Qian Liang and Zhenghui He contributed equally to this work and should be considered co-first authors.

Authors and Affiliations

  1. Brain Injury Center, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, People’s Republic of China

    Yuxiao Ma, Chun Yang, Zhenghui He, Jiyao Jiang & Junfeng Feng

  2. Shanghai Institute of Head Trauma, Shanghai, 200127, People’s Republic of China

    Yuxiao Ma, Chun Yang, Zhenghui He, Jiyao Jiang & Junfeng Feng

  3. Department of Pathology, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA

    Qian Liang

  4. Department of Biochemistry and Molecular Cell Biology, Shanghai Key Laboratory for Tumor Microenvironment and Inflammation, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China

    Weiji Weng

  5. Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People’s Republic of China

    Jin Lei

  6. Department of Neurosurgery, Yuquan Hospital Affiliated to Tsinghua University School of Clinical Medicine, Beijing, 100040, People’s Republic of China

    Loren Skudder-Hill

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  1. Yuxiao Ma
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Contributions

Yuxiao Ma, Chun Yang, Qian Liang, and Zhenghui He performed the experiments and wrote the manuscript. Weiji Weng designed the experiments and created the pictures. Jin Lei, Loren Skudder-Hill, and Jiyao Jiang edited the manuscript. Junfeng Feng designed and managed the entire study.

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Correspondence to Junfeng Feng.

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Supplementary Information

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12035_2022_2815_MOESM1_ESM.jpg

Supplement Figure 1: Network between ERK, GSK3β, cofilin, MAPK signaling pathway and actin cytoskeleton reorganization. Blue circles represent the gene and orange circles represent the pathway. The lines between genes and pathways represent the reported articles that investigated associations between the two genes or pathways (JPG 2275 KB)

12035_2022_2815_MOESM2_ESM.xlsx

Supplement Table 1: The differently expressed genes data from the RNA sequencing (absolute fold > 2, p < 0.05) between no DCEF and DCEF groups. (XLSX 741 KB)

12035_2022_2815_MOESM3_ESM.xlsx

Supplement Table 2: The up-regulated KEGG pathway enrichment analysis data between no DCEF and DCEF groups. (XLSX 49 KB)

Supplement Video 1: BV2 microglia migration video without DCEF. (MP4 2887 KB)

Supplement Video 2: BV2 microglia migration video under 150 mV/mm DCEF stimulation. (MP4 2960 KB)

Supplement Video 3: BV2 microglia migration video under 300 mV/mm DCEF stimulation. (MP4 3033 KB)

Supplement Video 4: BV2 microglia migration video with reversed DCEF stimulation at the middle of the time period. (MP4 3153 KB)

Supplement Video 5: BV2 microglia migration video under 300 mV/mm DCEF stimulation with 100 nM PMA. (MP4 1943 KB)

Supplement Video 6: BV2 microglia migration video under 300 mV/mm DCEF stimulation with 20 mM LiCl. (MP4 2.21 MB)

Supplement Video 7: BV2 microglia migration video under 300 mV/mm DCEF stimulation with cofilin siRNA. (MP4 2043 KB)

Supplement Video 8: BV2 microglia transfected with cofilin siRNA and cofilin(S3A) plasmid migration video under 300 mV/mm DCEF stimulation. (MP4 3031 KB)

Supplementary file12 (XLSX 7248 KB)

Supplementary file13 (AVI 9029 KB)

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Ma, Y., Yang, C., Liang, Q. et al. Direct Current Electric Field Coordinates the Migration of BV2 Microglia via ERK/GSK3β/Cofilin Signaling Pathway. Mol Neurobiol 59, 3665–3677 (2022). https://doi.org/10.1007/s12035-022-02815-5

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