Unlike the central nervous system, peripheral nerves can regenerate when damaged. MicroRNA (miRNA) is a novel class of small, non-coding RNA that regulates gene expression at the post-transcriptional level. Here, we report regular alterations of miRNA expression following rat sciatic nerve injury using deep sequencing. We harvested dorsal root ganglia tissues and the proximal stumps of the nerve, and identified 201 and 225 known miRNAs with significant expression variance at five time points in these tissues after sciatic nerve transaction, respectively. Subsequently, hierarchical clustering, miRNA expression pattern and co-expression network were performed. We screened out specific miRNAs and further obtained the intersection genes through target analysis software (Targetscan and miRanda). Moreover, GO and KEGG enrichment analyses of these intersection genes were performed. The bioinformatics analysis indicated that the potential targets for these miRNAs were involved in nerve regeneration, including neurogenesis, neuron differentiation, vesicle-mediated transport, homophilic cell adhesion and negative regulation of programmed cell death that were known to play important roles in regulating nerve repair. Finally, we combined differentially expressed mRNA with the predicted targets for selecting inverse miRNA-target pairs. Our results show that the abnormal expression of miRNA may contribute to illustrate the molecular mechanisms of nerve regeneration and that miRNAs are potential targets for therapeutic interventions and may enhance intrinsic regenerative ability.
References
[1]
Gruart A, Streppel M, Guntinas-Lichius O, Angelov DN, Neiss WF, et al. (2003) Motoneuron adaptability to new motor tasks following two types of facial-facial anastomosis in cats. Brain 126: 115–133.
[2]
Rishal I, Fainzilber M (2010) Retrograde signaling in axonal regeneration. Exp Neurol 223: 5–10.
[3]
Gu X, Ding F, Yang Y, Liu J (2011) Construction of tissue engineered nerve grafts and their application in peripheral nerve regeneration. Prog Neurobiol 93: 204–230.
[4]
Griffin JW, Pan B, Polley MA, Hoffman PN, Farah MH (2010) Measuring nerve regeneration in the mouse. Exp Neurol 223: 60–71.
[5]
Parrinello S, Napoli I, Ribeiro S, Digby PW, Fedorova M, et al. (2010) EphB signaling directs peripheral nerve regeneration through Sox2-dependent Schwann cell sorting. Cell 143: 145–155.
[6]
Fields RD, Stevens-Graham B (2002) New insights into neuron-glia communication. Science 298: 556–562.
[7]
Carthew RW, Sontheimer EJ (2009) Origins and Mechanisms of miRNAs and siRNAs. Cell 136: 642–655.
[8]
Friedman RC, Farh KK, Burge CB, Bartel DP (2009) Most mammalian mRNAs are conserved targets of microRNAs. Genome Res 19: 92–105.
Liu NK, Wang XF, Lu QB, Xu XM (2009) Altered microRNA expression following traumatic spinal cord injury. Exp Neurol 219: 424–429.
[12]
Nakanishi K, Nakasa T, Tanaka N, Ishikawa M, Yamada K, et al. (2010) Responses of microRNAs 124a and 223 following spinal cord injury in mice. Spinal Cord 48: 192–196.
[13]
Zhang HY, Zheng SJ, Zhao JH, Zhao W, Zheng LF, et al. (2011) MicroRNAs 144, 145, and 214 are down-regulated in primary neurons responding to sciatic nerve transection. Brain Res 1383: 62–70.
[14]
Zhou S, Yu B, Qian T, Yao D, Wang Y, et al. (2011) Early changes of microRNAs expression in the dorsal root ganglia following rat sciatic nerve transection. Neurosci Lett 494: 89–93.
[15]
Stark MS, Tyagi S, Nancarrow DJ, Boyle GM, Cook AL, et al. (2010) Characterization of the Melanoma miRNAome by Deep Sequencing. PLoS One 5: e9685.
[16]
Jagadeeswaran G, Zheng Y, Sumathipala N, Jiang H, Arrese EL, et al. (2010) Deep sequencing of small RNA libraries reveals dynamic regulation of conserved and novel microRNAs and microRNA-stars during silkworm development. BMC Genomics 11: 52.
[17]
Schulte JH, Marschall T, Martin M, Rosenstiel P, Mestdagh P, et al. (2010) Deep sequencing reveals differential expression of microRNAs in favorable versus unfavorable neuroblastoma. Nucleic Acids Res 38: 5919–5928.
[18]
Huang JC, Babak T, Corson TW, Chua G, Khan S, et al. (2007) Using expression profiling data to identify human microRNA targets. Nat Methods 4: 1045–1049.
[19]
Wang K, Li P, Dong Y, Cai X, Hou D, et al. (2011) A microarray-based approach identifies ADP ribosylation factor-like protein 2 as a target of microRNA-16. J Biol Chem 286: 9468–9476.
[20]
Navarro X, Vivó M, Valero-Cabré A (2007) Neural plasticity after peripheral nerve injury and regeneration. Prog Neurobiol 82: 163–201.
[21]
Rossi F, Gianola S, Corvetti L (2007) Regulation of intrinsic neuronal properties for axon growth and regeneration. Prog Neurobiol 81: 1–28.
[22]
Costigan M, Befort K, Karchewski L, Griffin RS, D'Urso D, et al. (2002) Replicate high-density rat genome oligonucleotide microarrays reveal hundreds of regulated genes in the dorsal root ganglion after peripheral nerve injury. BMC Neurosci 3: 16.
[23]
Xiao HS, Huang QH, Zhang FX, Bao L, Lu YJ, et al. (2002) Identification of gene expression profile of dorsal root ganglion in the rat peripheral axotomy model of neuropathic pain. Proc Natl Acad Sci U S A 99: 8360–8365.
[24]
Nagarajan R, Le N, Mahoney H, Araki T, Milbrandt J (2002) Deciphering peripheral nerve myelination by using Schwann cell expression profiling. Proc Natl Acad Sci U S A 99: 8998–9003.
[25]
Lewis BP, Burge CB, Bartel DP (2005) Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120: 15–20.
[26]
Krichevsky AM, Gabriely G (2009) MiR-21: a small multi-faceted RNA. J Cell Mol Med 13: 39–53.
[27]
Hoffman PN (2010) A conditioning lesion induces changes in gene expression and axonal transport that enhance regeneration by increasing the intrinsic growth state of axons. Exp Neurol 223: 11–18.
Braun S, Croizat B, Lagrange MC, Warter JM, Poindron P (1996) Neurotrophins increase motoneurons' ability to innervate skeletal muscle fibers in rat spinal cord–human muscle cocultures. J Neurol Sci 136: 17–23.
[30]
Oudega M, Hagg T (1999) Neurotrophins promote regeneration of sensory axons in the adult rat spinal cord. Brain Res 818: 431–438.
[31]
Zhang JY, Luo XG, Xian CJ, Liu ZH, Zhou XF (2000) Endogenous BDNF is required for myelination and regeneration of injured sciatic nerve in rodents. Eur J Neurosci 12: 4171–4180.
[32]
Mellios N, Huang HS, Grigorenko A, Rogaev E, Akbarian S (2008) A set of differentially expressed miRNAs, including miR-30a-5p, act as post-transcriptional inhibitors of BDNF in prefrontal cortex. Hum Mol Genet 17: 3030–3042.
[33]
Che YH, Yamashita T, Tohyama M (2002) Changes in mRNA for VAMPs following facial nerve transection. J Chem Neuroanat 24: 147–152.
[34]
Li XB, Fan XP, Wang XL, Cai L, Yang WC (2005) The cotton ACTIN1 gene is functionally expressed in fibers and participates in fiber elongation. Plant Cell 17: 859–875.
[35]
Redell JB, Zhao J, Dash PK (2011) Altered expression of miRNA-21 and its targets in the hippocampus after traumatic brain injury. J Neurosci Res 89: 212–221.
[36]
Martin DM, Skidmore JM, Philips ST, Vieira C, Gage PJ, et al. (2008) PITX2 is required for normal development of neurons in the mouse subthalamic nucleus and midbrain. Dev Biol 267: 93–108.
[37]
Avdonin PP, Grigorian EN, Markitantova IuV (2010) Transcriptional factor Pitx2: localization during triton retina regeneration. Izv Akad Nauk Ser Biol 3: 283–288.
[38]
Meng F, Henson R, Wehbe-Janek H, Ghoshal K, Jacob ST, et al. (2007) MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology 133: 647–658.
[39]
Christie KJ, Webber CA, Martinez JA, Singh B, Zochodne DW (2010) PTEN inhibition to facilitate intrinsic regenerative outgrowth of adult peripheral axons. J Neurosci 30: 9306–9315.
[40]
Liu K, Lu Y, Lee JK, Samara R, Willenberg R, et al. (2010) PTEN deletion enhances the regenerative ability of adult corticospinal neurons. Nat Neurosci 13: 1075–1081.
[41]
Park KK, Liu K, Hu Y, Smith PD, Wang C, et al. (2008) Promoting axon regeneration in the adult CNS by modulation of the PTEN/mTOR pathway. Science 322: 963–966.
[42]
Nave KA (2010) Myelination and support of axonal integrity by glia. Nature 468: 244–252.
[43]
Jessen KR, Mirsky R (2005) The origin and development of glial cells in peripheral nerves. Nat Rev Neurosci 6: 671–682.
[44]
Lee KH, Chen YL, Yeh SD, Hsiao M, Lin JT, et al. (2009) MicroRNA-330 acts as tumor suppressor and induces apoptosis of prostate cancer cells through E2F1-mediated suppression of Akt phosphorylation. Oncogene 28: 3360–3370.
[45]
Hirata H, Hibasami H, Yoshida T, Ogawa M, Matsumoto M, et al. (2001) Nerve growth factor signaling of p75 induces differentiation and ceramide-mediated apoptosis in Schwann cells cultured from degenerating nerves. Glia 36: 245–258.
[46]
Radstake TR, Gorlova O, Rueda B, Martin JE, Alizadeh BZ, et al. (2010) Genome-wide association study of systemic sclerosis identifies CD247 as a new susceptibility locus. Nat Genet 42: 426–429.
[47]
Wang H, Lu Y, Rudd CE (2010) SKAP1 is dispensable for chemokine-induced migration of primary T-cells. Immunol Lett 128: 148–153.
[48]
Volakakis N, Kadkhodaei B, Joodmardi E, Wallis K, Panman L, et al. (2010) NR4A orphan nuclear receptors as mediators of CREB-dependent neuroprotection. Proc Natl Acad Sci U S A 107: 12317–12322.
[49]
Blin M, Norton W, Bally-Cuif L, Vernier P (2008) NR4A2 controls the differentiation of selective dopaminergic nuclei in the zebrafish brain. Mol Cell Neurosci 39: 592–604.
[50]
Magnani P, Cherian PV, Gould GW, Greene DA, Sima AA, et al. (1996) Glucose transporters in rat peripheral nerve: paranodal expression of GLUT1 and GLUT3. Metabolism 45: 1466–1473.
[51]
Wayman GA, Davare M, Ando H, Fortin D, Varlamova O, et al. (2008) An activity-regulated microRNA controls dendritic plasticity by downregulating p250GAP. Proc Natl Acad Sci U S A 105: 9093–9098.
[52]
Edbauer D, Neilson JR, Foster KA, Wang CF, Seeburg DP, et al. (2010) Regulation of synaptic structure and function by FMRP-associated microRNAs miR-125b and miR-132. Neuron 65: 373–384.