Neural Regeneration Research ›› 2018, Vol. 13 ›› Issue (12): 2182-2190.doi: 10.4103/1673-5374.241469

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Analysis of transcriptome sequencing of sciatic nerves in Sprague-Dawley rats of different ages

Jiang-Hui Liu1, Qing Tang2, Xiang-Xia Liu2, Jian Qi3, Rui-Xi Zeng2, Zhao-Wei Zhu2, Bo He4, Yang-Bin Xu2   

  1. 1 Department of Emergency, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
    2 Department of Plastic Surgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
    3 Department of Orthopedics and Microsurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
    4 Department of Orthopedics, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
  • Received:2018-08-20 Online:2018-12-15 Published:2018-12-15
  • Contact: Zhao-Wei Zhu, MD, PhD, nmtmemoir@aliyun.com; Bo He, MD, PhD, hebodoc@gmail.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81201546 (to YXL); the Doctoral Start-up Program of Natural Science Foundation of Guangdong Province of China, No. 2017A030310302 (to ZWZ); the Medical Scientific Research Foundation of Guangdong Province of China, No. A2016018 (to BH); the Science and Technology Project of Guangdong Province of China, No.2016A010103012 (to JHL).

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Abstract:

An aging-induced decrease in Schwann cell viability can affect regeneration following peripheral nerve injury in mammals. It is therefore necessary to investigate possible age-related changes in gene expression that may affect the biological function of peripheral nerves. Ten 1-week-old and ten 12-month-old healthy male Sprague-Dawley rats were divided into young (1 week old) and adult (12 months old) groups according to their ages. mRNA expression in the sciatic nerve was compared between young and adult rats using next-generation sequencing (NGS) and bioinformatics (n = 4/group). The 18 groups of differentially expressed mRNA (DEmRNAs) were also tested by quantitative reverse transcription polymerase chain reaction (n = 6/group). Results revealed that (1) compared with young rats, adult rats had 3608 groups of DEmRNAs. Of these, 2684 were groups of upregulated genes, and 924 were groups of downregulated genes. Their functions mainly involved cell viability, proliferation, differentiation, regeneration, and myelination. (2) The gene with the most obvious increase of all DEmRNAs in adult rats was Thrsp (log2FC = 9.01, P < 0.05), and the gene with the most obvious reduction was Col2a1 (log2FC = –8.89, P < 0.05). (3) Gene Ontology analysis showed that DEmRNAs were mainly concentrated in oligosaccharide binding, nucleotide-binding oligomerization domain containing one signaling pathway, and peptide-transporting ATPase activity. (4) Analysis using the Kyoto Encyclopedia of Genes and Genomes showed that, with increased age, DEmRNAs were mainly enriched in steroid biosynthesis, Staphylococcus aureus infection, and graft-versus-host disease. (5) Spearman’s correlation coefficient method for evaluating NGS accuracy showed that the NGS results and quantitative reverse transcription polymerase chain reaction results were positively correlated (rs = 0.74, P < 0.05). These findings confirm a difference in sciatic nerve gene expression between adult and young rats, suggesting that, in peripheral nerves, cells and the microenvironment change with age, thus influencing the function and repair of peripheral nerves.

Key words: peripheral nerve injury, aging, Sprague-Dawley rat, transcriptome, sequencing, mRNA, rat age, Schwann cells, neural regeneration