Exploration of molecular pathways mediating electric field-directed schwann cell migration by RNA-seq

Abstract

In peripheral nervous systems, Schwann cells wrap around axons of motor and sensory neurons to form the myelin sheath. Following spinal cord injury, Schwann cells regenerate and migrate to the lesion and are involved in the spinal cord regeneration process. Transplantation of Schwann cells into injured neural tissue results in enhanced spinal axonal regeneration. Effective directional migration of Schwann cells is critical in the neural regeneration process. In this study, we report that Schwann cells migrate anodally in an applied electric field (EF). The directedness and displacement of anodal migration increased significantly when the strength of the EF increased from 50 mV/mm to 200 mV/mm. The EF did not significantly affect the cell migration speed. To explore the genes and signaling pathways that regulate cell migration in EFs, we performed a comparative analysis of differential gene expression between cells stimulated with an EF (100 mV/mm) and those without using next-generation RNA sequencing, verified by RT-qPCR. Based on the cut-off criteria (FC > 1.2, q < 0.05), we identified 1,045 up-regulated and 1,636 down-regulated genes in control cells versus EF-stimulated cells. A Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis found that compared to the control group, 21 pathways are down-regulated, while 10 pathways are up-regulated. Differentially expressed genes participate in multiple cellular signaling pathways involved in the regulation of cell migration, including pathways of regulation of actin cytoskeleton, focal adhesion, and PI3K-Akt.

SREBP and MDT-15 protect C. elegans from glucose-induced accelerated aging by preventing accumulation of saturated fat

“…GCF_000002985.6]) by Bowtie2 (Langmead and Salzberg 2012), and differentially expressed genes were analyzed using the edgeR method (Robinson et al 2010) by the CLRNASeq program (ChunLab, Inc.). Genes with significant changes in expression (P < 0.05, fold change >1.2) (Yao et al 2015) were further analyzed, and genes whose FPKM (fragments per kilobase of transcript per million fragments) was zero were excluded. Scatter plot (Riedel et al 2013) and heat map analyses (Ewald et al 2015) were performed as previously described.…”

Genetic analysis of the molecular regulation of electric fields-guided glia migration

“…The effect of EFs on cell migration has been reported in a number of investigations. In previous studies, we found that both SCs 18 and OPCs migrated toward the anodal pole of an applied EF 19 . The increase of directedness of anodal migration of these cells is in proportion to the strength of the applied EF (50 to 200 mV/mm).…”

“…Signal transduction is involved in the assembly and disassembly of the protein complex thereby controlling cell motility and migration direction. In previous publications, we have reported that the migration of SCs and OPCs can be directed by an applied electric field, and we performed RNA-seq to explore the pathways that regulated the cell migration for each type 18,19 . In this study, we jointly analyzed the RNA-seq data for OPCs and SCs subjected to EFs and then identified significantly enriched GO terms associated with DEGs.…”

“…The RNA-seq reads from OPCs and Schwann cells exposed to electrical fields were previously described in the work of Yao et al 18 and Li et al 19 , respectively. The OPCs and Schwann cells that were used in those studies were isolated from cerebral cortexes of postnatal rats (days 1-2) and the sciatic nerves of postnatal rats (days 1-3) respectively.…”

“…Exploring the gene expression in neural cells subjected to EF stimulation may also lead to the improvement of strategies to regulate neural cell migration. www.nature.com/scientificreports/ conclusion In summary, in this study, we performed comparative differential expression analysis for RNA-seq reads from OPCs and SCs that were subjected to EF stimulation in our previous reports in Yao et al 18 and Li et al 19 , respectively. GO term analysis of the DEGs revealed a number of significantly enriched GO terms related to cytoskeleton, cell adhesion, cell movement, and cell migration.…”