Histochemistry and Cell Biology

, Volume 147, Issue 3, pp 307–316 | Cite as

High mobility group box 1 (HMGB1): dual functions in the cochlear auditory neurons in response to stress?

  • Sabine Ladrech
  • Jing Wang
  • Marc Mathieu
  • Jean-Luc Puel
  • Marc Lenoir
Original Paper


High mobility group box 1 (HMGB1) is a DNA-binding protein that facilitates gene transcription and may act extracellularly as a late mediator of inflammation. The roles of HMGB1 in the pathogenesis of the spiral ganglion neurons (SGNs) of the cochlea are currently unknown. In the present study, we tested the hypothesis that early phenotypical changes in the SGNs of the amikacin-poisoned rat cochlea are mediated by HMGB1. Our results showed that a marked downregulation of HMGB1 had occurred by completion of amikacin treatment, coinciding with acute damage at the dendrite extremities of the SGNs. A few days later, during the recovery of the SGN dendrites, the protein was re-expressed and transiently accumulated within the nuclei of the SGNs. The phosphorylated form of the transcription factor c-Jun (p-c-Jun) was concomitantly detected in the nuclei of the SGNs where it often co-localized with HMGB1, while the anti-apoptotic protein BCL2 was over-expressed in the cytoplasm. In animals co-treated with amikacin and the histone deacetylase inhibitor trichostatin A, both HMGB1 and p-c-Jun were exclusively found within the cytoplasm. The initial disappearance of HMGB1 from the affected SGNs may be due to its release into the external medium, where it may have a cytokine-like function. Once re-expressed and translocated into the nucleus, HMGB1 may facilitate the transcriptional activity of p-c-Jun, which in turn may promote repair mechanisms. Our study therefore suggests that HMGB1 can positively influence the survival of SGNs following ototoxic exposure via both its extracellular and intranuclear functions.


Excitotoxicity Ototoxicity c-Jun Gamma-H2ax BCL2 TSA 



We acknowledge Julien Menardo and Romain Lalandes for their helpful advice during the course of the experiments. Thanks are due to Hassan Boukhaddaoui for assistance in fluorescent imagery (technological platform Montpellier Rio Imaging (MRI)), and to Chantal Cazevieille and Alicia Caballero for assistance in electron microscopy (technological platform Correlative Microscopy and Electron Tomography (COMET)) at the Institute of Neurosciences of Montpellier (INM). Cryosections were made in the Experimental Histology Network of Montpellier (RHEM) of the INM. Experimental animals were given attentive care by the animal facility team of INM. The French Ministère de la Recherche et des Nouvelles Technologies provided financial support for cell imagery. The manuscript has been revised for the English by an independent scientific language editing service (Angloscribe, 30420-Calvisson, France).

Supplementary material

418_2016_1506_MOESM1_ESM.tif (1.2 mb)
Supplementary material 1 (TIFF 1268 kb)


  1. Agresti A, Bianchi ME (2003) HMGB proteins and gene expression. Curr Opin Genet Dev 13:170–178CrossRefPubMedGoogle Scholar
  2. Bianchi ME, Manfredi AA (2014) How macrophages ring the inflammation alarm. Proc Natl Acad Sci USA 111:2866–2867CrossRefPubMedPubMedCentralGoogle Scholar
  3. Bichler E, Spoendlin H, Rauchegger H (1983) Degeneration of cochlear neurons after amikacin intoxication in the rat. Arch Otorhinolaryngol 237:201–208CrossRefPubMedGoogle Scholar
  4. Bonaldi T, Talamo F, Scaffidi P, Ferrera D, Porto A, Bachi A, Rubartelli A, Agresti A, Bianchi ME (2003) Monocytic cells hyperacetylate chromatin protein HMGB1 to redirect it towards secretion. EMBO J 22:5551–5560CrossRefPubMedPubMedCentralGoogle Scholar
  5. Budenz CL, Pfingst BE, Raphael Y (2012) The use of neurotrophin therapy in the inner ear to augment cochlear implantation outcomes. Anat Rec (Hoboken) 295:1896–1908CrossRefGoogle Scholar
  6. Campana L, Bosurgi L, Bianchi ME, Manfredi AA, Rovere-Querini P (2009) Requirement of HMGB1 for stromal cell-derived factor-1/CXCL12-dependent migration of macrophages and dendritic cells. J Leukocyte Biol 86:609–615CrossRefPubMedGoogle Scholar
  7. Chen C-J, Chang W-C, Ben-Kuen Chen B-K (2008) Attenuation of c-Jun and Sp1 expression and p300 recruitment to gene promoter confers the trichostatin A-induced inhibition of 12(S)-lipoxygenase expression in EGF-treated A431 cells. Eur J Pharmacol 591:36–42CrossRefPubMedGoogle Scholar
  8. Chen L, Dean C, Gandolfi M, Nahm E, Mattiace L, Kim AH (2014) Dexamethasone’s effect in the retrocochlear auditory centers of a noise-induced hearing loss mouse model. Otolaryngol Head Neck Surg 151:667–674CrossRefPubMedGoogle Scholar
  9. De Toma I, Rossetti G, Zambrano S, Bianchi ME, Agresti A (2014) Nucleosome loss facilitates the chemotactic response of macrophages. J Intern Med 276:454–469CrossRefPubMedGoogle Scholar
  10. Faraco G, Fossati S, Bianchi ME, Patrone M, Pedrazzi M, Sparatore B, Moroni F, Chiarugi A (2007) High mobility group box 1 protein is released by neural cells upon different stresses and worsens ischemic neurodegeneration in vitro and in vivo. J Neurochem 103:590–603CrossRefPubMedGoogle Scholar
  11. Felder E, Schrott-Fischer A (1995) Quantitative evaluation of myelinated nerve fibres and hair cells in cochleae of humans with age-related high-tone hearing loss. Hear Res 91:19–32CrossRefPubMedGoogle Scholar
  12. Finnin MS, Donigian JR, Cohen A, Richon VM, Rifkind RA, Marks PA, Breslow R, Pavletich NP (1999) Structures of a histone deacetylase homologue bound to the TSA and SAHA inhibitors. Nature 401:188–193CrossRefPubMedGoogle Scholar
  13. Gillespie LN, Shepherd RK (2005) Clinical application of neurotrophic factors: the potential for primary auditory neuron protection. Eur J Neurosci 22:2123–2133CrossRefPubMedPubMedCentralGoogle Scholar
  14. Hansen MR, Roehm PC, Xu N, Green SH (2007) Overexpression of Bcl-2 or Bcl-xL prevents spiral ganglion neuron death and inhibits neurite growth. Dev Neurobiol 67:316–325CrossRefPubMedGoogle Scholar
  15. Herdegen T, Skene P, Bähr M (1997) The c-Jun transcription factor-bipotential mediator of neuronal death, survival and regeneration. Trends Neurosci 20:227–231CrossRefPubMedGoogle Scholar
  16. Joshi SR, Sarpong YC, Peterson RC, Scovell WM (2012) Nucleosome dynamics: HMGB1 relaxes canonical nucleosome structure to facilitate estrogen receptor binding. Nucleic Acids Res 40:10161–10171CrossRefPubMedPubMedCentralGoogle Scholar
  17. Kaur T, Zamani D, Tong L, Rubel EW, Ohlemiller KK, Hirose K, Warchol ME (2015) Fractalkine signaling regulates macrophage recruitment into the cochlea and promotes the survival of spiral ganglion neurons after selective hair cell lesion. J Neurosci 35:15050–15061CrossRefPubMedPubMedCentralGoogle Scholar
  18. Kim JB, Lim CM, Yu YM, Lee JK (2008) Induction and subcellular localization of high-mobility group box-1 (HMGB1) in the postischemic rat brain. J Neurosci Res 86:1125–1131CrossRefPubMedGoogle Scholar
  19. Kujawa SG, Liberman MC (2006) Acceleration of age-related hearing loss by early noise exposure: evidence of a misspent youth. J Neurosci 26:2115–2123CrossRefPubMedPubMedCentralGoogle Scholar
  20. Kujawa SG, Liberman MC (2009) Adding insult to injury: cochlear nerve degeneration after “temporary” noise-induced hearing loss. J Neurosci 29:14077–14085CrossRefPubMedPubMedCentralGoogle Scholar
  21. Kujawa SG, Liberman MC (2015) Synaptopathy in the noise-exposed and aging cochlea: primary neural degeneration in acquired sensorineural hearing loss. Hear Res 330:191–199CrossRefPubMedPubMedCentralGoogle Scholar
  22. Kuo LJ, Yang LX (2008) Gamma-H2AX—a novel biomarker for DNA double-strand breaks. In Vivo 22:305–309PubMedGoogle Scholar
  23. Ladrech S, Guitton M, Saido T, Lenoir M (2004) Calpain activity in the amikacin-damaged rat cochlea. J Comp Neurol 477:149–160CrossRefPubMedGoogle Scholar
  24. Ladrech S, Wang J, Simonneau L, Puel J-L, Lenoir M (2007) Macrophage contribution to the response of the rat organ of Corti to amikacin. J Neurosci Res 85:1970–1979CrossRefPubMedGoogle Scholar
  25. Ladrech S, Mathieu M, Puel JL, Lenoir M (2013) Supporting cells regulate the remodelling of aminoglycoside-injured organ of Corti, through the release of high mobility group box 1. Eur J Neurosci 38:2962–2972PubMedGoogle Scholar
  26. Lallemend F, Lefebvre PP, Hans G, Rigo JM, Van de Water TR, Moonen G, Malgrange B (2003) Substance P protects spiral ganglion neurons from apoptosis via PKC-Ca2 + -MAPK/ERK pathways. J Neurochem 87:508–521CrossRefPubMedGoogle Scholar
  27. Lallemend F, Hadjab S, Hans G, Moonen G, Lefebvre PP, Malgrange B (2005) Activation of protein kinase CbetaI constitutes a new neurotrophic pathway for deafferented spiral ganglion neurons. J Cell Sci 118:4511–4525CrossRefPubMedGoogle Scholar
  28. Lang H, Schulte BA, Zhou D, Smythe N, Spicer SS, Schmiedt RA (2006) Nuclear factor κB deficiency is associated with auditory nerve degeneration and increased noise-induced hearing loss. J Neurosci 26:3541–3550CrossRefPubMedPubMedCentralGoogle Scholar
  29. Lange SS, Mitchell DL, Vasquez KM (2008) High mobility group protein B1 enhances DNA repair and chromatin modification after DNA damage. Proc Natl Acad Sci USA 105:10320–10325CrossRefPubMedPubMedCentralGoogle Scholar
  30. Lanuszewska J, Widlak P (2000) High mobility group 1 and 2 proteins bind preferentially to DNA that contains bulky adducts induced by benzopyrene diol epoxide and N-acetoxy-acetylaminofluorene. Cancer Lett 158:17–25CrossRefPubMedGoogle Scholar
  31. Le Prell CG, Yagi M, Kawamoto K, Beyer LA, Atkin G, Raphael Y, Dolan DF, Bledsoe SC Jr, Moody DB (2004) Chronic excitotoxicity in the guinea pig cochlea induces temporary functional deficits without disrupting otoacoustic emissions. J Acoust Soc Am 116:1044–1056CrossRefPubMedGoogle Scholar
  32. Lenoir M, Daudet N, Humbert G, Renard N, Gallego M, Pujol R, Eybalin M, Vago P (1999) Morphological and molecular changes in the inner hair cell region of the cochlea after amikacin treatment. J Neurocytol 28:925–937CrossRefPubMedGoogle Scholar
  33. Li G, Liu W, Frenz D (2006) Cisplatin ototoxicity to the rat inner ear: a role for HMG1 and iNOS. Neurotoxicology 27:22–30CrossRefPubMedGoogle Scholar
  34. Lin HW, Furman AC, Kujawa SG, Liberman MC (2011) Primary neural degeneration in the Guinea pig cochlea after reversible noise-induced threshold shift. J Assoc Res Otolaryngol 12:605–616CrossRefPubMedPubMedCentralGoogle Scholar
  35. Lindwall C, Kanje M (2005) The role of p-c-Jun in survival and outgrowth of developing sensory neurons. NeuroReport 16:1655–1659CrossRefPubMedGoogle Scholar
  36. Liu Y, Prasad R, Wilson SH (2010) HMGB1: roles in base excision repair and related function. Biochim Biophys Acta 1799:119–130CrossRefPubMedPubMedCentralGoogle Scholar
  37. Pasheva EA, Pashev IG, Favre A (1998) Preferential binding of high mobility group 1 protein to UV-damaged DNA. Role of the COOH-terminal domain. J Biol Chem 273:24730–24736CrossRefPubMedGoogle Scholar
  38. Puel JL, d’Aldin C, Ruel J, Ladrech S, Pujol R (1997) Synaptic repair mechanisms responsible for functional recovery in various cochlear pathologies. Acta Otolaryngol 117:214–218CrossRefPubMedGoogle Scholar
  39. Pujol R, Puel JL (1999) Excitotoxicity, synaptic repair, and functional recovery in the mammalian cochlea: a review of recent findings. Ann N Y Acad Sci 884:249–254CrossRefPubMedGoogle Scholar
  40. Raivich G (2008) c-Jun expression, activation and function in neural cell death, inflammation and repair. J Neurochem 107:898–906PubMedGoogle Scholar
  41. Ramekers D, Versnel H, Grolman W, Klis S (2012) Neurotrophins and their role in the cochlea. Hear Res 288:19–33CrossRefPubMedGoogle Scholar
  42. Reeves R, Adair JE (2005) Role of high mobility group (HMG) chromatin proteins in DNA repair. DNA Repair 4:926–938CrossRefPubMedGoogle Scholar
  43. Ridder DA, Schwaninger M (2009) NF-κB signaling in cerebral ischemia. Neuroscience 158:995–1006CrossRefPubMedGoogle Scholar
  44. Robertson D (1983) Functional significance of dendritic swelling after loud sounds in the guinea pig cochlea. Hear Res 9:263–278CrossRefPubMedGoogle Scholar
  45. Rowell JP, Simpson KL, Stott K, Watson M, Thomas JO (2012) HMGB1-facilitated p53 DNA binding occurs via HMG-Box/p53 transactivation domain interaction, regulated by the acidic tail. Structure 20:2014–2024CrossRefPubMedGoogle Scholar
  46. Ruel J, Wang J, Rebillard G, Eybalin M, Lloyd R, Pujol R, Puel JL (2007) Physiology, pharmacology and plasticity at the inner hair cell synaptic complex. Hear Res 227:19–27CrossRefPubMedGoogle Scholar
  47. Scaffidi P, Misteli T, Bianchi ME (2002) Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature 418:191–195CrossRefPubMedGoogle Scholar
  48. Siddiqui WA, Ahad A, Ahsan H (2015) The mystery of BCL2 family: Bcl-2 proteins and apoptosis: an update. Arch Toxicol 89:289–317CrossRefPubMedGoogle Scholar
  49. Spoendlin H (1971) Primary structural changes in the organ of Corti after acoustic overstimulation. Acta Otolaryngol 71:166–176CrossRefPubMedGoogle Scholar
  50. Spoendlin H (1985) Anatomy of cochlear innervation. Am J Otolaryngol 6:453–467CrossRefPubMedGoogle Scholar
  51. Takahashi A, Ohnishi T (2005) Does gH2AX foci formation depend on the presence of DNA double strand breaks? Cancer Lett 229:171–179CrossRefPubMedGoogle Scholar
  52. Tang D, Kang R, Livesey KM, Zeh HJ, Lotze MT (2011) High mobility group box 1 (HMGB1) activates an autophagic response to oxidative stress. Antioxid Redox Signal 15:2185–2195CrossRefPubMedPubMedCentralGoogle Scholar
  53. Tournier C, Hess P, Yang DD, Xu J, Turner TK, Nimnual A, Bar-Sagi D, Jones SN, Flavell RA, Davis RJ (2000) Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. Science 288:870–887CrossRefPubMedGoogle Scholar
  54. Travers AA (2003) Priming the nucleosome: a role for HMGB proteins? EMBO Rep 4:131–136CrossRefPubMedPubMedCentralGoogle Scholar
  55. Vandenbosch R, Chocholova E, Robe PA, Wang Y, Lambert C, Moonen G, Lallemend F, Malgrange B, Hadjab S (2013) A role for the canonical nuclear factor-κB pathway in coupling neurotrophin-induced differential survival of developing spiral ganglion neurons. Front Cell Neurosci 7:242CrossRefPubMedPubMedCentralGoogle Scholar
  56. Vénéreau E, Ceriotti C, Bianchi ME (2015) DAMPs from cell death to new life. Front Immunol 6:422CrossRefPubMedPubMedCentralGoogle Scholar
  57. Wong AC, Ryan AF (2015) Mechanisms of sensorineural cell damage, death and survival in the cochlea. Front Aging Neurosci 7:58PubMedPubMedCentralGoogle Scholar
  58. Wu Q, Zhang W, Pwee KH, Kumar PP (2003) Rice HMGB1 protein recognizes DNA structures and bends DNA efficiently. Arch Biochem Biophys 411:105–111CrossRefPubMedGoogle Scholar
  59. Wyllie AH (2010) “Where, O death, is thy sting?” A brief review of apoptosis biology. Mol Neurobiol 42:4–9CrossRefPubMedPubMedCentralGoogle Scholar
  60. Yamaguchi K, Lantowski A, Dannenberg AJ, Subbaramaiah K (2005) Histone deacetylase inhibitors suppress the induction of c-Jun and its target genes including COX-2. J Biol Chem 280:32569–32577CrossRefPubMedGoogle Scholar
  61. Yarza R, Vela S, Solas M, Ramirez MJ (2016) c-Jun N-terminal Kinase (JNK) Signaling as a Therapeutic Target for Alzheimer’s Disease. Front Pharmacol 6:321CrossRefPubMedPubMedCentralGoogle Scholar
  62. Zuccotti A, Kuhn S, Johnson SL, Franz C, Singer W, Hecker D, Geisler HS, Köpschall I, Rohbock K, Gutsche K, Dlugaiczyk J, Schick B, Marcotti W, Rüttiger L, Schimmang T, Knipper M (2012) Lack of brain-derived neurotrophic factor hampers inner hair cell synapse physiology, but protects against noise-induced hearing loss. J Neurosci 32:8545–8553CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Sabine Ladrech
    • 1
    • 3
  • Jing Wang
    • 1
    • 3
  • Marc Mathieu
    • 2
    • 3
  • Jean-Luc Puel
    • 1
    • 3
  • Marc Lenoir
    • 1
    • 3
  1. 1.INSERM, U1051, Institute for Neurosciences (INM)Hôpital Saint EloiMontpellier Cedex 5France
  2. 2.INSERM, U1183, Institute of Regenerative Medicine and Biotherapy (IRMB)MontpellierFrance
  3. 3.University of MontpellierMontpellierFrance

Personalised recommendations