Advertisement

Cellular and Molecular Neurobiology

, Volume 39, Issue 1, pp 87–98 | Cite as

MiR-20a Plays a Key Regulatory Role in the Repair of Spinal Cord Dorsal Column Lesion via PDZ-RhoGEF/RhoA/GAP43 Axis in Rat

  • Tianyi Wang
  • Bo Li
  • Xin Yuan
  • Libin Cui
  • Zhijie Wang
  • Yanjun Zhang
  • Mei Yu
  • Yucai Xiu
  • Zheng Zhang
  • Wenhua Li
  • Fengyan Wang
  • Xiaoling GuoEmail author
  • Xiangyang ZhaoEmail author
  • Xueming ChenEmail author
Original Research
  • 69 Downloads

Abstract

Spinal cord injury (SCI) causes sensory dysfunctions such as paresthesia, dysesthesia, and chronic neuropathic pain. MiR-20a facilitates the axonal outgrowth of the cortical neurons. However, the role of miR-20a in the axonal outgrowth of primary sensory neurons and spinal cord dorsal column lesion (SDCL) is yet unknown. Therefore, the role of miR-20a post-SDCL was investigated in rat. The NF-200 immunofluorescence staining was applied to observe whether axonal outgrowth of dorsal root ganglion (DRG) neurons could be altered by miR-20a or PDZ-RhoGEF modulation in vitro. The expression of miR-20a was quantized with RT-PCR. Western blotting analyzed the expression of PDZ-RhoGEF/RhoA/GAP43 axis after miR-20a or PDZ-RhoGEF was modulated. The spinal cord sensory conduction function was assessed by somatosensory-evoked potentials and tape removal test. The results demonstrated that the expression of miR-20a decreased in a time-dependent manner post-SDCL. The regulation of miR-20a modulated the axonal growth and the expression of PDZ-RhoGEF/RhoA/GAP43 axis in vitro. The in vivo regulation of miR-20a altered the expression of miR-20a-PDZ-RhoGEF/RhoA/GAP43 axis and promoted the recovery of ascending sensory function post-SDCL. The results indicated that miR-20a/PDZ-RhoGEF/RhoA/GAP43 axis is associated with the pathophysiological process of SDCL. Thus, targeting the miR-20a/PDZ-RhoGEF /RhoA/GAP43 axis served as a novel strategy in promoting the sensory function recovery post-SCI.

Keywords

MicroRNA-20a Dorsal column lesion PDZ-RhoGEF RhoA Neurite growth Dorsal root ganglion Primary sensory neuron 

Notes

Acknowledgements

This work was supported by the General Program of Natural Science Foundation of Hebei Province of China (Grant Number H2017101030), the Medical Science and Technology Youth Cultivation Project of the Chinese People’s Liberation Army (Grant Numbers 16QNP074 and 13QNP017), the Research and Development of Science and Technology Program Supported by Chengde Government (Grant Number 201606A062), the Beijing Municipal Education Commission General Project (KM201710025028), the Capital characteristic project (Z161100000116064), and the Public Experiment Training Foundation of Beijing Luhe Hospital, Capital Medical University, China (Grant Number lh201425Shi).

Author Contributions

XG, XZ and XC designed this study; ZW was in charge of the methodology; LC and MY were in charge of the software and validation; Formal analysis was done by YX; ZZ and WL were in charge of the investigation; FW was in charge of data curation; BL wrote the original draft; TW reviewed and edited the draft; XY was in charge of the visualization; YZ and XY supervised the study; TW administrated this study; Funding was acquired from TW; ZW, YZ and XC.

Compliance with Ethical Standards

Conflict of interest

The authors declare no conflict of interest.

Ethical Approval

ARRIVE guidelines and National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Publications no. 85-23, revised 1996) for the care and use of animals were followed. All protocols performed in this study were approved by the Ethics Committee of the 266th Hospital of the Chinese People’s Liberation Army (Approval No. 20170038).

References

  1. Ahmed Z, Douglas MR, Read ML, Berry M, Logan A (2011) Citron kinase regulates axon growth through a pathway that converges on cofilin downstream of RhoA. Neurobiol Dis 41:421–429.  https://doi.org/10.1016/j.nbd.2010.10.012 CrossRefGoogle Scholar
  2. Al-Chalabi M, Alsalman I (2018) Neuroanatomy, posterior column (dorsal column). In: StatPearls. StatPearls Publishing LLC, Treasure Island (FL)Google Scholar
  3. Attwell CL, van Zwieten M, Verhaagen J, Mason MRJ (2018) The dorsal column lesion model of spinal cord injury and its use in deciphering the neuron-intrinsic injury response. Dev Neurobiol.  https://doi.org/10.1002/dneu.22601 Google Scholar
  4. Bhalala OG, Pan L, Sahni V, Mcguire TL, Gruner K, Tourtellotte WG, Kessler JA (2012) microRNA-21 regulates astrocytic response following spinal cord injury. J Neurosci 32:17935–17947CrossRefGoogle Scholar
  5. Caizhong X, Chunlei S, Beibei L, Zhiqing D, Qinneng D, Tong W (2014) The application of somatosensory evoked potentials in spinal cord injury rehabilitation. Neuro Rehabil 35:835–840  https://doi.org/10.3233/NRE-141158 Google Scholar
  6. Cirillo G, Colangelo AM, De Luca C, Savarese L, Barillari MR, Alberghina L, Papa M (2016) Modulation of matrix metalloproteinases activity in the ventral horn of the spinal cord re-stores neuroglial synaptic homeostasis and neurotrophic support following peripheral nerve injury. PLoS ONE 11:e0152750.  https://doi.org/10.1371/journal.pone.0152750 CrossRefGoogle Scholar
  7. Eva R, Koseki H, Kanamarlapudi V, Fawcett JW (2017) EFA6 regulates selective polarised transport and axon regeneration from the axon initial segment. J Cell Sci 130:3663–3675.  https://doi.org/10.1242/jcs.207423 CrossRefGoogle Scholar
  8. Fagoe ND, Attwell CL, Eggers R, Tuinenbreijer L, Kouwenhoven D, Verhaagen J, Mason MR (2016) Evaluation of five tests for sensitivity to functional deficits following cervical or thoracic dorsal column transection in the rat. PLoS ONE 11:e0150141.  https://doi.org/10.1371/journal.pone.0150141 CrossRefGoogle Scholar
  9. Gao L, Dai C, Feng Z, Zhang L, Zhang Z (2018) MiR-137 inhibited inflammatory response and apoptosis after spinal cord injury via targeting of MK2. J Cell Biochem 119:3280–3292CrossRefGoogle Scholar
  10. Goganau I, Sandner B, Weidner N, Fouad K, Blesch A (2017) Depolarization and electrical stimulation enhance in vitro and in vivo sensory axon growth after spinal cord injury. Exp Neurol 300:247CrossRefGoogle Scholar
  11. Han X et al (2011) Simvastatin treatment improves functional recovery after experimental spinal cord injury by upregulating the expression of BDNF and GDNF. Neurosci Lett 487:255–259.  https://doi.org/10.1016/j.neulet.2010.09.007 CrossRefGoogle Scholar
  12. Hebert SS, Horre K, Nicolai L, Bergmans B, Papadopoulou AS, Delacourte A, De Strooper B (2009) MicroRNA regulation of Alzheimer’s Amyloid precursor protein expression. Neurobiol Dis 33:422–428.  https://doi.org/10.1016/j.nbd.2008.11.009 CrossRefGoogle Scholar
  13. Hoschouer EL, Finseth T, Flinn S, Basso DM, Jakeman LB (2010) Sensory stimulation prior to spinal cord injury induces post-injury dysesthesia in mice. J Neurotrauma 27:777–787.  https://doi.org/10.1089/neu.2009.1182 CrossRefGoogle Scholar
  14. Hu J, Zhang G, Rodemer W, Jin LQ, Shifman M, Selzer ME (2016) The role of RhoA in retrograde neuronal death and axon regeneration after spinal cord injury. Neurobiol Dis 98:25–35CrossRefGoogle Scholar
  15. Jia L, Chopp M, Wang L, Lu X, Zhang Y, Szalad A, Zhang ZG (2018) MiR-34a regulates axonal growth of dorsal root ganglia neurons by targeting FOXP2 and VAT1 in postnatal and adult mouse. Mol Neurobiol  https://doi.org/10.1007/s12035-018-1047-3 Google Scholar
  16. Kaas JH, Qi HX, Burish MJ, Gharbawie OA, Onifer SM, Massey JM (2008) Cortical and subcortical plasticity in the brains of humans, primates, and rats after damage to sensory afferents in the dorsal columns of the spinal cord. Exp Neurol 209:407–416.  https://doi.org/10.1016/j.expneurol.2007.06.014 CrossRefGoogle Scholar
  17. Ko HR et al (2016) Akt1-Inhibitor of DNA binding2 is essential for growth cone formation and axon growth and promotes central nervous system axon regeneration. Elife 5:e20799  https://doi.org/10.7554/eLife.20799 CrossRefGoogle Scholar
  18. Krishnan KR, Glass CA, Turner SM, Watt JW, Fraser MH (1992) Perceptual deprivation in the acute phase of spinal injury rehabilitation. J Am Parapleg Soc 15:60–65CrossRefGoogle Scholar
  19. Kuner R, Swiercz JM, Zywietz A, Tappe A, Offermanns S (2002) Characterization of the expression of PDZ-RhoGEF, LARG and G(alpha)12/G(alpha)13 proteins in the murine nervous system. Eur J Neurosci 16:2333–2341CrossRefGoogle Scholar
  20. Kwon MJ et al (2013) Contribution of macrophages to enhanced regenerative capacity of dorsal root ganglia sensory neurons by conditioning injury. J Neurosci 33:15095–15108.  https://doi.org/10.1523/JNEUROSCI.0278-13.2013 CrossRefGoogle Scholar
  21. Lin MY, Lin YM, Kao TC, Chuang HH, Chen RH (2011) PDZ-RhoGEF ubiquitination by Cullin3-KLHL20 controls neurotrophin-induced neurite outgrowth. J Cell Biol 193:985–994.  https://doi.org/10.1083/jcb.201103015 CrossRefGoogle Scholar
  22. Liu D, Huang Y, Jia C, Li Y, Liang F, Fu Q (2015) Administration of antagomir-223 inhibits apoptosis, promotes angiogenesis and functional recovery in rats with spinal cord injury. Cell Mol Neurobiol 35:483–491.  https://doi.org/10.1007/s10571-014-0142-x CrossRefGoogle Scholar
  23. Mason MR et al (2010) Comparison of AAV serotypes for gene delivery to dorsal root ganglion neurons. Mol Ther 18:715CrossRefGoogle Scholar
  24. McKinley W, Santos K, Meade M, Brooke K (2007) Incidence and outcomes of spinal cord injury clinical syndromes. J Spinal Cord Med 30:215–224CrossRefGoogle Scholar
  25. Moreno-Flores MT et al (2006) A clonal cell line from immortalized olfactory ensheathing glia promotes functional recovery in the injured spinal cord. Mol Ther 13:598–608.  https://doi.org/10.1016/j.ymthe.2005.11.014 CrossRefGoogle Scholar
  26. Neumann S, Woolf CJ (1999) Regeneration of dorsal column fibers into and beyond the lesion site following adult spinal cord injury. Neuron 23:83–91CrossRefGoogle Scholar
  27. Niederost B, Oertle T, Fritsche J, McKinney RA, Bandtlow CE (2002) Nogo-A and myelin-associated glycoprotein mediate neurite growth inhibition by antagonistic regulation of RhoA and Rac1. J Neurosci 22:10368–10376CrossRefGoogle Scholar
  28. Okada S, Hara M, Kobayakawa K, Matsumoto Y, Nakashima Y (2018) Astrocyte reactivity and astrogliosis after spinal cord injury. Neurosci Res 126:39–43.  https://doi.org/10.1016/j.neures.2017.10.004 CrossRefGoogle Scholar
  29. Papa M, De Luca C, Petta F, Alberghina L, Cirillo G (2014) Astrocyte-neuron interplay in maladaptive plasticity. Neurosci Biobehav Rev 42:35–54.  https://doi.org/10.1016/j.neubiorev.2014.01.010 CrossRefGoogle Scholar
  30. Qiu J, Cafferty WB, McMahon SB, Thompson SW (2005) Conditioning injury-induced spinal axon regeneration requires signal transducer and activator of transcription 3 activation. J Neurosci 25:1645–1653.  https://doi.org/10.1523/JNEUROSCI.3269-04.2005 CrossRefGoogle Scholar
  31. Reuther GW et al (2001) Leukemia-associated Rho guanine nucleotide exchange factor, a Dbl family protein found mutated in leukemia, causes transformation by activation of RhoA. J Biol Chem 276:27145–27151.  https://doi.org/10.1074/jbc.M103565200 CrossRefGoogle Scholar
  32. Richards JS, Meredith RL, Nepomuceno C, Fine PR, Bennett G (1980) Psycho-social aspects of chronic pain in spinal cord injury. Pain 8:355–366CrossRefGoogle Scholar
  33. Richards JS, Hirt M, Melamed L (1982) Spinal cord injury: a sensory restriction perspective. Arch Phys Med Rehabil 63:195–199Google Scholar
  34. Ropper AE, Ropper AH (2017) Acute Spinal Cord Compression N. Engl J Med 376:1358–1369CrossRefGoogle Scholar
  35. Shang X et al (2013) Small-molecule inhibitors targeting G-protein-coupled Rho guanine nucleotide exchange factors. Proc Natl Acad Sci USA 110:3155–3160.  https://doi.org/10.1073/pnas.1212324110 CrossRefGoogle Scholar
  36. Sun X, Zhou Z, Fink DJ, Mata M (2013) HspB1 silences translation of PDZ-RhoGEF by enhancing miR-20a and miR-128 expression to promote neurite extension. Mol Cell Neurosci 57:111–119.  https://doi.org/10.1016/j.mcn.2013.10.006 CrossRefGoogle Scholar
  37. Wang E, Cambi F (2012) MicroRNA expression in mouse oligodendrocytes and regulation of proteolipid protein gene expression. J Neurosci Res 90:1701–1712.  https://doi.org/10.1002/jnr.23055 CrossRefGoogle Scholar
  38. Wang W et al (2014) SNAP25 ameliorates sensory deficit in rats with spinal cord transection. Mol Neurobiol 50:290–304CrossRefGoogle Scholar
  39. Wang T et al (2015a) miR-142-3p is a potential therapeutic target for sensory function recovery of spinal cord injury medical science monitor international medical. J Exp Clin Res 21:2553–2556Google Scholar
  40. Wang Y, Cui H, Pu J, Luk KDK, Hu Y (2015b) Time-frequency patterns of somatosensory evoked potentials in predicting the location of spinal cord injury. Neurosci Lett 603:37–41.  https://doi.org/10.1016/j.neulet.2015.07.002 CrossRefGoogle Scholar
  41. Wang Z et al (2018) PEITC promotes neurite growth in Primary sensory neurons via the miR-17-5p/STAT3/GAP-43 Axis J Drug Target.  https://doi.org/10.1080/1061186x.2018.1486405 Google Scholar
  42. Xie W et al (2018) Knockdown of MicroRNA-21 promotes neurological recovery after acute spinal cord injury. Neurochem Res  https://doi.org/10.1007/s11064-018-2580-1 Google Scholar
  43. Xu J, He J, He H, Peng R, Xi J (2016) Comparison of RNAi NgR and NEP1-40 in acting on axonal regeneration after spinal cord injury in rat. Models Mol Neurobiol 54:1–11Google Scholar
  44. Yang G, Tang W-Y (2017) Resistance of interleukin-6 to the extracellular inhibitory environment promotes axonal regeneration and functional recovery following spinal cord injury. Int J Mol Med 39(2):437–445CrossRefGoogle Scholar
  45. Zhang G, Lei F, Zhou Q, Feng D, Bai Y (2016) Combined application of Rho-ROCKII and GSK-3β inhibitors exerts an improved protective effect on axonal regeneration in rats with spinal cord injury Molecular Medicine Reports 14Google Scholar
  46. Zheng Y (2001) Dbl family guanine nucleotide exchange factors. Trends Biochem sci 26:724–732CrossRefGoogle Scholar
  47. Zhou S et al (2012) microRNA-222 targeting PTEN promotes neurite outgrowth from adult dorsal root ganglion neurons following sciatic nerve transection. PLoS ONE 7:e44768.  https://doi.org/10.1371/journal.pone.0044768 CrossRefGoogle Scholar
  48. Zhou Y et al (2014) Matrix metalloproteinase-1 (MMP-1) expression in rat spinal cord injury model. Cell Mol Neurobiol 34:1151–1163.  https://doi.org/10.1007/s10571-014-0090-5 CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of OrthopedicsThe 266th Hospital of the Chinese People’s Liberation ArmyChengdePeople’s Republic of China
  2. 2.Department of OrthopedicsTianjin Medical University General HospitalTianjinPeople’s Republic of China
  3. 3.Department of Spine Surgery, Beijing Luhe HospitalCapital Medical UniversityBeijingPeople’s Republic of China
  4. 4.Department of Pediatric Internal MedicineAffiliated Hospital of Chengde Medical UniversityChengdePeople’s Republic of China
  5. 5.Leukemia Center, Peking Union of Medical College, Institute of Hematology & Hospital of Blood DiseasesChinese Academy of Medical SciencesTianjinPeople’s Republic of China
  6. 6.Department of NeurologyThe 266th Hospital of the Chinese People’s Liberation ArmyChengdePeople’s Republic of China
  7. 7.Department of General SurgeryThe 266th Hospital of the Chinese People’s Liberation ArmyChengdePeople’s Republic of China

Personalised recommendations