Expression and localization of diacylglycerol kinase ζ in guinea pig cochlea and its functional implication under noise-exposure stress conditions

  • Chikako Shinkawa
  • Tsukasa ItoEmail author
  • Yasukazu Hozumi
  • Makoto Chiba
  • Hirooki Matsui
  • Kaoru Goto
  • Seiji Kakehata
Original Paper


Cochlear hair cells are essential for the mechanotransduction of hearing. Sensorineural hearing loss can be irreversible because hair cells have a minimal ability to repair or regenerate themselves once damaged. In order to develop therapeutic interventions to prevent hair cell loss, it is necessary to understand the signaling pathway operating in cochlear hair cells and its alteration upon damage. Diacylglycerol kinase (DGK) regulates intracellular signal transduction through phosphorylation of lipidic second messenger diacylglycerol. We have previously reported characteristic expression and localization patterns of DGKs in various organs under pathophysiological conditions. Nevertheless, little is known about morphological and functional aspects of this enzyme family in the cochlea. First RT-PCR analysis reveals predominant mRNA expression of DGKα, DGKε and DGKζ. Immunohistochemical analysis shows that DGKζ localizes to the nuclei of inner hair cells (IHCs), outer hair cells (OHCs), supporting cells and spiral ganglion neurons in guinea pig cochlea under normal conditions. It is well known that loud noise exposure induces cochlear damage, thereby resulting in hair cell loss. In particular, OHCs are highly vulnerable to noise exposure than IHCs. We found that after 1 week of noise exposure DGKζ translocates from the nucleus to the cytoplasm in damage-sensitive OHCs and gradually disappears thereafter. In sharp contrast, DGKζ remains to the nucleus in damage-resistant IHCs. These results suggest that DGKζ cytoplasmic translocation is well correlated with cellular damage under noise-exposure stress conditions and is involved in delayed cell death in cochlear outer hair cells.


Diacylglycerol kinase Noise exposure Sensorineural hearing loss Outer hair cell Cytoplasmic translocation 



Diacylglycerol kinase


Inner hair call


Outer hair cell


Supporting cell


Spiral ganglion neuron



This work was supported by JSPS KAKENHI Grant Numbers JP15K20178 to HM, JP15K20179 to CS and JP17K11313 to TI.


  1. Ali H, Nakano T, Saino-Saito S, Hozumi Y, Katagiri Y, Kamii H, Sato S, Kayama T, Kondo H, Goto K (2004) Selective translocation of diacylglycerol kinase zeta in hippocampal neurons under transient forebrain ischemia. Neurosci Lett 372:190–195. CrossRefGoogle Scholar
  2. Bunting M, Tang W, Zimmerman GA, McIntyre TM, Prescott SM (1996) Molecular cloning and characterization of a novel human diacylglycerol kinase zeta. J Biol Chem 271:10230–10236CrossRefGoogle Scholar
  3. Ding L, Traer E, McIntyre TM, Zimmerman GA, Prescott SM (1998) The cloning and characterization of a novel human diacylglycerol kinase, DGKiota. J Biol Chem 273:32746–32752CrossRefGoogle Scholar
  4. Evangelisti C, Astolfi A, Gaboardi GC, Tazzari P, Pession A, Goto K, Martelli AM (2009) TIS21/BTG2/PC3 and cyclin D1 are key determinants of nuclear diacylglycerol kinase-zeta-dependent cell cycle arrest. Cell Signal 21:801–809CrossRefGoogle Scholar
  5. Evangelisti C, Gaboardi GC, Billi AM, Ognibene A, Goto K, Tazzari PL, McCubrey JA, Martelli AM (2010) Identification of a functional nuclear export sequence in diacylglycerol kinase-zeta. Cell Cycle 9:384–388. CrossRefGoogle Scholar
  6. Forge A, Li L, Corwin JT, Nevill G (1993) Ultrastructural evidence for hair cell regeneration in the mammalian inner ear. Science 259:1616–1619CrossRefGoogle Scholar
  7. Fujioka M, Kanzaki S, Okano HJ, Masuda M, Ogawa K, Okano H (2006) Proinflammatory cytokines expression in noise-induced damaged cochlea. J Neurosci Res 83:575–583. CrossRefGoogle Scholar
  8. Goto K, Kondo H (1993) Molecular cloning and expression of a 90-kDa diacylglycerol kinase that predominantly localizes in neurons. Proc Natl Acad Sci USA 90:7598–7602CrossRefGoogle Scholar
  9. Goto K, Kondo H (1996) A 104-kDa diacylglycerol kinase containing ankyrin-like repeats localizes in the cell nucleus. Proc Natl Acad Sci USA 93:11196–11201CrossRefGoogle Scholar
  10. Goto K, Funayama M, Kondo H (1994) Cloning and expression of a cytoskeleton-associated diacylglycerol kinase that is dominantly expressed in cerebellum. Proc Natl Acad Sci USA 91:13042–13046CrossRefGoogle Scholar
  11. Goto K, Hozumi Y, Nakano T, Saino SS, Kondo H (2007) Cell biology and pathophysiology of the diacylglycerol kinase family: morphological aspects in tissues and organs. Int Rev Cytol 264:25–63. CrossRefGoogle Scholar
  12. Goto K, Tanaka T, Nakano T, Okada M, Hozumi Y, Topham MK, Martelli AM (2014) DGKzeta under stress conditions: “To be nuclear or cytoplasmic, that is the question”. Adv Biol Regul 54:242–253. CrossRefGoogle Scholar
  13. Herrup K, Yang Y (2007) Cell cycle regulation in the postmitotic neuron: oxymoron or new biology? Nat Rev Neurosci 8:368–378. CrossRefGoogle Scholar
  14. Houssa B, Schaap D, van der Wal J, Goto K, Kondo H, Yamakawa A, Shibata M, Takenawa T, van Blitterswijk WJ (1997) Cloning of a novel human diacylglycerol kinase (DGKtheta) containing three cysteine-rich domains, a proline-rich region, and a pleckstrin homology domain with an overlapping Ras-associating domain. J Biol Chem 272:10422–10428CrossRefGoogle Scholar
  15. Hozumi Y, Ito T, Nakano T, Nakagawa T, Aoyagi M, Kondo H, Goto K (2003) Nuclear localization of diacylglycerol kinase zeta in neurons. Eur J Neurosci 18:1448–1457CrossRefGoogle Scholar
  16. Hozumi Y, Fukaya M, Adachi N, Saito N, Otani K, Kondo H, Watanabe M, Goto K (2008) Diacylglycerol kinase beta accumulates on the perisynaptic site of medium spiny neurons in the striatum. Eur J Neurosci 28:2409–2422. CrossRefGoogle Scholar
  17. Hozumi Y, Watanabe M, Goto K (2010) Signaling cascade of diacylglycerol kinase beta in the pituitary intermediate lobe: dopamine D2 receptor/phospholipase Cbeta4/diacylglycerol kinase beta/protein kinase Calpha. J Histochem Cytochem 58:119–129. CrossRefGoogle Scholar
  18. Hozumi Y, Matsui H, Sakane F, Watanabe M, Goto K (2013) Distinct expression and localization of diacylglycerol kinase isozymes in rat retina. J Histochem Cytochem 61:462–476. CrossRefGoogle Scholar
  19. Hozumi Y, Akimoto R, Suzuki A, Otani K, Watanabe M, Goto K (2015) Expression and localization of the diacylglycerol kinase family and of phosphoinositide signaling molecules in adrenal gland. Cell Tissue Res 362:295–305. CrossRefGoogle Scholar
  20. Hozumi Y, Nakano T, Tanaka T, Goto K (2016) Localization of diacylglycerol kinase zeta in rat pancreatic islet cells under normal and streptozotocin-induced stress conditions. Arch Histol Cytol 76:23–33. CrossRefGoogle Scholar
  21. Hozumi Y, Fujiwara H, Kaneko K, Fujii S, Topham MK, Watanabe M, Goto K (2017) Diacylglycerol kinase epsilon localizes to subsurface cisterns of cerebellar Purkinje cells. Cell Tissue Res 368:441–458. CrossRefGoogle Scholar
  22. Imai S, Kai M, Yasuda S, Kanoh H, Sakane F (2005) Identification and characterization of a novel human type II diacylglycerol kinase, DGK kappa. J Biol Chem 280:39870–39881. CrossRefGoogle Scholar
  23. Ito T, Hozumi Y, Sakane F, Saino-Saito S, Kanoh H, Aoyagi M, Kondo H, Goto K (2004) Cloning and characterization of diacylglycerol kinase iota splice variants in rat brain. J Biol Chem 279:23317–23326. CrossRefGoogle Scholar
  24. Jamesdaniel S, Hu B, Kermany MH, Jiang H, Ding D, Coling D, Salvi R (2011) Noise induced changes in the expression of p38/MAPK signaling proteins in the sensory epithelium of the inner ear. J Proteomics 75:410–424. CrossRefGoogle Scholar
  25. Kai M, Sakane F, Imai S, Wada I, Kanoh H (1994) Molecular cloning of a diacylglycerol kinase isozyme predominantly expressed in human retina with a truncated and inactive enzyme expression in most other human cells. J Biol Chem 269:18492–18498Google Scholar
  26. Kanoh H, Yamada K, Sakane F (1990) Diacylglycerol kinase: a key modulator of signal transduction? Trends Biochem Sci 15:47–50CrossRefGoogle Scholar
  27. Kirino T (1982) Delayed neuronal death in the gerbil hippocampus following ischemia. Brain Res 239:57–69CrossRefGoogle Scholar
  28. Lau A, Tymianski M (2010) Glutamate receptors, neurotoxicity and neurodegeneration. Pflugers Arch 460:525–542. CrossRefGoogle Scholar
  29. Los AP, Vinke FP, de Widt J, Topham MK, van Blitterswijk WJ, Divecha N (2006) The retinoblastoma family proteins bind to and activate diacylglycerol kinase zeta. J Biol Chem 281:858–866. CrossRefGoogle Scholar
  30. Maeda Y, Fukushima K, Omichi R, Kariya S, Nishizaki K (2013) Time courses of changes in phospho- and total- MAP kinases in the cochlea after intense noise exposure. PLoS One 8:e58775. CrossRefGoogle Scholar
  31. Martelli AM, Fala F, Faenza I, Billi AM, Cappellini A, Manzoli L, Cocco L (2004) Metabolism and signaling activities of nuclear lipids. Cell Mol Life Sci 61:1143–1156. CrossRefGoogle Scholar
  32. Merida I, Avila-Flores A, Merino E (2008) Diacylglycerol kinases: at the hub of cell signalling. Biochem J 409:1–18. CrossRefGoogle Scholar
  33. Miller JM, Brown JN, Schacht J (2003) 8-Iso-prostaglandin F(2alpha), a product of noise exposure, reduces inner ear blood flow. Audiol Neurootol 8:207–221. CrossRefGoogle Scholar
  34. Mizutari K, Fujioka M, Hosoya M, Bramhall N, Okano HJ, Okano H, Edge AS (2013) Notch inhibition induces cochlear hair cell regeneration and recovery of hearing after acoustic trauma. Neuron 77:58–69. CrossRefGoogle Scholar
  35. Murakami T, Sakane F, Imai S, Houkin K, Kanoh H (2003) Identification and characterization of two splice variants of human diacylglycerol kinase eta. J Biol Chem 278:34364–34372. CrossRefGoogle Scholar
  36. Nishizuka Y (1992) Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 258:607–614CrossRefGoogle Scholar
  37. Okada M, Hozumi Y, Tanaka T, Suzuki Y, Yanagida M, Araki Y, Evangelisti C, Yagisawa H, Topham MK, Martelli AM, Goto K (2012) DGKzeta is degraded through the cytoplasmic ubiquitin-proteasome system under excitotoxic conditions, which causes neuronal apoptosis because of aberrant cell cycle reentry. Cell Signal 24:1573–1582. CrossRefGoogle Scholar
  38. Oshima K, Shin K, Diensthuber M, Peng AW, Ricci AJ, Heller S (2010) Mechanosensitive hair cell-like cells from embryonic and induced pluripotent stem cells. Cell 141:704–716. CrossRefGoogle Scholar
  39. Quirk WS, Seidman MD (1995) Cochlear vascular changes in response to loud noise. Am J Otol 16:322–325Google Scholar
  40. Rhee SG, Suh PG, Ryu SH, Lee SY (1989) Studies of inositol phospholipid-specific phospholipase C. Science 244:546–550CrossRefGoogle Scholar
  41. Ron D, Kazanietz MG (1999) New insights into the regulation of protein kinase C and novel phorbol ester receptors. FASEB journal: FASEB J 13:1658–1676CrossRefGoogle Scholar
  42. Saino-Saito S, Hozumi Y, Goto K (2011) Excitotoxicity by kainate-induced seizure causes diacylglycerol kinase zeta to shuttle from the nucleus to the cytoplasm in hippocampal neurons. Neurosci Lett 494:185–189. CrossRefGoogle Scholar
  43. Sakane F, Kanoh H, Yokoyama C, Tanabe T (1990) Porcine diaglycerol kinase sequence has zinc finger and E-F hand motifs. Nature 344:345–348CrossRefGoogle Scholar
  44. Sakane F, Imai S, Yamada K, Murakami T, Tsushima S, Kanoh H (2002) Alternative splicing of the human diacylglycerol kinase delta gene generates two isoforms differing in their expression patterns and in regulatory functions. J Biol Chem 277:43519–43526. CrossRefGoogle Scholar
  45. Sakane F, Imai S, Kai M, Yasuda S, Kanoh H (2007) Diacylglycerol kinases: why so many of them? Biochim Biophys Acta 1771:793–806. CrossRefGoogle Scholar
  46. Sakane F, Mizuno S, Komenoi S (2016) Diacylglycerol kinases as emerging potential drug targets for a variety of diseases: an update. Front Cell Dev Biol 4:82. CrossRefGoogle Scholar
  47. Schaap D, de Widt J, van der Wal J, Vandekerckhove J, van Damme J, Gussow D, Ploegh HL, van Blitterswijk WJ, van der Bend RL (1990) Purification, cDNA-cloning and expression of human diacylglycerol kinase. FEBS Lett 275:151–158. CrossRefGoogle Scholar
  48. Schnedl WJ, Ferber S, Johnson JH, Newgard CB (1994) STZ transport and cytotoxicity. Specific enhancement in GLUT2-expressing cells. Diabetes 43:1326–1333CrossRefGoogle Scholar
  49. Slepecky NB, Ulfendahl M (1993) Evidence for calcium-binding proteins and calcium-dependent regulatory proteins in sensory cells of the organ of Corti. Hear Res 70:73–84CrossRefGoogle Scholar
  50. Sulg M, Kirjavainen A, Pajusola K, Bueler H, Ylikoski J, Laiho M, Pirvola U (2010) Differential sensitivity of the inner ear sensory cell populations to forced cell cycle re-entry and p53 induction. J Neurochem 112:1513–1526. CrossRefGoogle Scholar
  51. Suzuki Y, Yamazaki Y, Hozumi Y, Okada M, Tanaka T, Iseki K, Ohta N, Aoyagi M, Fujii S, Goto K (2012) NMDA receptor-mediated Ca(2+) influx triggers nucleocytoplasmic translocation of diacylglycerol kinase zeta under oxygen-glucose deprivation conditions, an in vitro model of ischemia, in rat hippocampal slices. Histochem Cell Biol 137:499–511. CrossRefGoogle Scholar
  52. Tan WJ, Thorne PR, Vlajkovic SM (2016) Characterisation of cochlear inflammation in mice following acute and chronic noise exposure. Histochem Cell Biol 146:219–230. CrossRefGoogle Scholar
  53. Tanaka T, Okada M, Hozumi Y, Tachibana K, Kitanaka C, Hamamoto Y, Martelli AM, Topham MK, Iino M, Goto K (2013) Cytoplasmic localization of DGKzeta exerts a protective effect against p53-mediated cytotoxicity. J Cell Sci 126:2785–2797. CrossRefGoogle Scholar
  54. Tang W, Bunting M, Zimmerman GA, McIntyre TM, Prescott SM (1996) Molecular cloning of a novel human diacylglycerol kinase highly selective for arachidonate-containing substrates. J Biol Chem 271:10237–10241CrossRefGoogle Scholar
  55. Topham MK (2006) Signaling roles of diacylglycerol kinases. J Cell Biochem 97:474–484. CrossRefGoogle Scholar
  56. Warchol ME, Lambert PR, Goldstein BJ, Forge A, Corwin JT (1993) Regenerative proliferation in inner ear sensory epithelia from adult guinea pigs and humans. Science 259:1619–1622CrossRefGoogle Scholar
  57. Yamashita D, Jiang HY, Schacht J, Miller JM (2004) Delayed production of free radicals following noise exposure. Brain Res 1019:201–209. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Otolaryngology, Head and Neck SurgeryYamagata University Faculty of MedicineYamagata-shiJapan
  2. 2.Department of Cell Biology and MorphologyAkita University Graduate School of MedicineAkitaJapan
  3. 3.Department of Anatomy and Cell BiologyYamagata University Faculty of MedicineYamagataJapan

Personalised recommendations