Advertisement

Cellular and Molecular Neurobiology

, Volume 34, Issue 6, pp 881–893 | Cite as

Regulatory Role of the JNK-STAT1/3 Signaling in Neuronal Differentiation of Cultured Mouse Embryonic Stem Cells

  • Zheng Zachory Wei
  • Shan Ping Yu
  • Jin Hwan Lee
  • Dongdong Chen
  • Tammi M. Taylor
  • Todd Carter Deveau
  • Albert Cheung Hoi Yu
  • Ling WeiEmail author
Original Research

Abstract

Stem cell transplantation therapy has provided promising hope for the treatment of a variety of neurodegenerative disorders. Among challenges in developing disease-specific stem cell therapies, identification of key regulatory signals for neuronal differentiation is an essential and critical issue that remains to be resolved. Several lines of evidence suggest that JNK, also known as SAPK, is involved in neuronal differentiation and neural plasticity. It may also play a role in neurite outgrowth during neuronal development. In cultured mouse embryonic stem (ES) cells, we test the hypothesis that the JNK pathway is required for neuronal differentiation. After neural induction, the cells were plated and underwent differentiation for up to 5 days. Western blot analysis showed a dramatic increase in phosphorylated JNKs at 1–5 days after plating. The phosphorylation of JNK subsequently induced activation of STAT1 and STAT3 that lead to expressions of GAP-43, neurofilament, βIII-tubulin, and synaptophysin. NeuN-colabelled with DCX, a marker for neuroblast, was enhanced by JNK signaling. Neuronal differentiation of ES cells was attenuated by treatment with SP600125, which inhibited the JNK activation and decreased the activation of STAT1 and STAT3, and consequently suppressed the expressions of GAP-43, neurofilament, βIII-tubulin, and the secretion of VEGF. Data from immunocytochemistry indicated that the nuclear translocation of STAT3 was reduced, and neurites of ES-derived neurons were shorter after treatment with SP600125 compared with control cells. These results suggest that the JNK-STAT3 pathway is a key regulator required for early neuronal differentiation of mouse ES cells. Further investigation on expression of JNK isoforms showed that JNK-3 was significantly upregulated during the differentiation stage, while JNK-1 and JNK-2 levels decreased. Our study provided interesting information on JNK functions during ES cell neuronal differentiation.

Keywords

Embryonic stem cell JNK STAT3 Neurite outgrowth Neuronal differentiation 

Notes

Acknowledgments

This work was supported by grants from the National Institutes of Health, USA (NS045810 to SPY, NS057255 and NS075338 to LW), the American Heart Association Established Investigator Award (0840110N to LW), a Grant-in-Aid award (12GRNT12060222 to SPY) and a VA national merit grant (SPY). This work was also supported by the NIH grant C06 RR015455 from the Extramural Research Facilities Program of the National Center for Research Resources.

Conflict of interest

All authors have no conflict of interest in this investigation.

References

  1. Alter J, Rozentzweig D, Bengal E (2008) Inhibition of myoblast differentiation by tumor necrosis factor alpha is mediated by c-Jun N-terminal kinase 1 and leukemia inhibitory factor. J Biol Chem 283(34):23224–23234. doi: 10.1074/801379200 PubMedCrossRefGoogle Scholar
  2. Anjomshoa M, Karbalaie K, Mardani M, Razavi S, Tanhaei S, Nasr-Esfahani MH, Baharvand H (2009) Generation of motor neurons by coculture of retinoic acid-pretreated embryonic stem cells with chicken notochords. Stem Cells Dev 18(2):259–267. doi: 10.1089/2008.0049 PubMedCrossRefGoogle Scholar
  3. Atkinson PJ, Cho CH, Hansen MR, Green SH (2011) Activity of all JNK isoforms contributes to neurite growth in spiral ganglion neurons. Hear Res 278(1–2):77–85. doi: 10.1016/2011.04.011 PubMedCentralPubMedCrossRefGoogle Scholar
  4. Barnat M, Enslen H, Propst F, Davis RJ, Soares S, Nothias F (2010) Distinct roles of c-Jun N-terminal kinase isoforms in neurite initiation and elongation during axonal regeneration. J Neurosci 30(23):7804–7816. doi: 10.1523/0372-10.2010 PubMedCrossRefGoogle Scholar
  5. Bennett BL, Sasaki DT, Murray BW, O’Leary EC, Sakata ST, Xu W, Leisten JC, Motiwala A, Pierce S, Satoh Y, Bhagwat SS, Manning AM, Anderson DW (2001) SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc Natl Acad Sci USA 98(24):13681–13686. doi: 10.1073/251194298 PubMedCentralPubMedCrossRefGoogle Scholar
  6. Byun K, Kim TK, Oh J, Bayarsaikhan E, Kim D, Lee MY, Pack CG, Hwang D, Lee B (2013) Heat shock instructs hESCs to exit from the self-renewal program through negative regulation of OCT4 by SAPK/JNK and HSF1 pathway. Stem Cell Res 11(3):1323–1334. doi: 10.1016/2013.08.014 PubMedCrossRefGoogle Scholar
  7. Cai B, Li X, Wang Y, Liu Y, Yang F, Chen H, Yin K, Tan X, Zhu J, Pan Z, Wang B, Lu Y (2013) Apoptosis of bone marrow mesenchymal stem cells caused by homocysteine via activating JNK signal. PLoS ONE 8(5):e63561. doi: 10.1371/0063561 PubMedCentralPubMedCrossRefGoogle Scholar
  8. Chang L, Kamata H, Solinas G, Luo JL, Maeda S, Venuprasad K, Liu YC, Karin M (2006) The E3 ubiquitin ligase itch couples JNK activation to TNFalpha-induced cell death by inducing c-FLIP(L) turnover. Cell 124(3):601–613. doi: 10.1016/2006.01.021 PubMedCrossRefGoogle Scholar
  9. Cui J, Wang Q, Wang J, Lv M, Zhu N, Li Y, Feng J, Shen B, Zhang J (2009) Basal c-Jun NH2-terminal protein kinase activity is essential for survival and proliferation of T-cell acute lymphoblastic leukemia cells. Mol Cancer Ther 8(12):3214–3222. doi: 10.1158/1535-7163 PubMedCrossRefGoogle Scholar
  10. Engberg N, Kahn M, Petersen DR, Hansson M, Serup P (2010) Retinoic acid synthesis promotes development of neural progenitors from mouse embryonic stem cells by suppressing endogenous, Wnt-dependent nodal signaling. Stem Cells 28(9):1498–1509. doi: 10.1002/479 PubMedCrossRefGoogle Scholar
  11. Eom DS, Choi WS, Oh YJ (2004) Bcl-2 enhances neurite extension via activation of c-Jun N-terminal kinase. Biochem Biophys Res Commun 314(2):377–381PubMedCrossRefGoogle Scholar
  12. Eom DS, Choi WS, Ji S, Cho JW, Oh YJ (2005) Activation of c-Jun N-terminal kinase is required for neurite outgrowth of dopaminergic neuronal cells. NeuroReport 16(8):823–828PubMedCrossRefGoogle Scholar
  13. Fanger GR, Gerwins P, Widmann C, Jarpe MB, Johnson GL (1997) MEKKs, GCKs, MLKs, PAKs, TAKs, and tpls: upstream regulators of the c-Jun amino-terminal kinases? Curr Opin Genet Dev 7(1):67–74PubMedCrossRefGoogle Scholar
  14. Fraser L, Taylor AH, Forrester LM (2013) SCF/KIT inhibition has a cumulative but reversible effect on the self-renewal of embryonic stem cells and on the survival of differentiating cells. Cell Reprogr 15(4):259–268. doi: 10.1089/2013.0015 Google Scholar
  15. Gdalyahu A, Ghosh I, Levy T, Sapir T, Sapoznik S, Fishler Y, Azoulai D, Reiner O (2004) DCX, a new mediator of the JNK pathway. EMBO J 23(4):823–832PubMedCentralPubMedCrossRefGoogle Scholar
  16. Guerrini R, Marini C (2006) Genetic malformations of cortical development. Exp Brain Res 173(2):322–333. doi: 10.1007/s00221-006-0501 PubMedCrossRefGoogle Scholar
  17. Haeusgen W, Boehm R, Zhao Y, Herdegen T, Waetzig V (2009) Specific activities of individual c-Jun N-terminal kinases in the brain. Neuroscience 161(4):951–959. doi: 10.1016/2009.04.014 PubMedCrossRefGoogle Scholar
  18. Haeusgen W, Herdegen T, Waetzig V (2011) MKK7gamma1 reverses nerve growth factor signals: proliferation and cell death instead of neuritogenesis and protection. Cell Signal 23(8):1281–1290. doi: 10.1016/2011.03.009 PubMedCrossRefGoogle Scholar
  19. He JC, Gomes I, Nguyen T, Jayaram G, Ram PT, Devi LA, Iyengar R (2005) The G alpha(o/i)-coupled cannabinoid receptor-mediated neurite outgrowth involves Rap regulation of Src and Stat3. J Biol Chem 280(39):33426–33434. doi: 10.1074/502812200 PubMedCrossRefGoogle Scholar
  20. Himes SR, Sester DP, Ravasi T, Cronau SL, Sasmono T, Hume DA (2006) The JNK are important for development and survival of macrophages. J Immunol 176(4):2219–2228PubMedCrossRefGoogle Scholar
  21. Holm KH, Cicchetti F, Bjorklund L, Boonman Z, Tandon P, Costantini LC, Deacon TW, Huang X, Chen DF, Isacson O (2001) Enhanced axonal growth from fetal human bcl-2 transgenic mouse dopamine neurons transplanted to the adult rat striatum. Neuroscience 104(2):397–405PubMedCrossRefGoogle Scholar
  22. Hui L, Zatloukal K, Scheuch H, Stepniak E, Wagner EF (2008) Proliferation of human HCC cells and chemically induced mouse liver cancers requires JNK1-dependent p21 downregulation. J Clin Investig 118(12):3943–3953. doi: 10.1172/37156 PubMedCentralPubMedCrossRefGoogle Scholar
  23. Kook SH, Jeon YM, Lim SS, Jang MJ, Cho ES, Lee SY, Choi KC, Kim JG, Lee JC (2013) Fibroblast growth factor-4 enhances proliferation of mouse embryonic stem cells via activation of c-Jun signaling. PLoS ONE 8(8):e71641. doi: 10.1371/0071641 PubMedCentralPubMedCrossRefGoogle Scholar
  24. Li Z, Theus MH, Wei L (2006) Role of ERK 1/2 signaling in neuronal differentiation of cultured embryonic stem cells. Dev Growth Differ 48(8):513–523. doi: 10.1111/1440-169X.2006.00889 PubMedCrossRefGoogle Scholar
  25. Liu X, Ye R, Yan T, Yu SP, Wei L, Xu G, Fan X, Jiang Y, Stetler RA, Liu G, Chen J (2013) Cell based therapies for ischemic stroke: from basic science to bedside. Prog Neurobiol. doi: 10.1016/2013.11.007 PubMedCentralGoogle Scholar
  26. Ming GL, Wong ST, Henley J, Yuan XB, Song HJ, Spitzer NC, Poo MM (2002) Adaptation in the chemotactic guidance of nerve growth cones. Nature 417(6887):411–418. doi: 10.1038/745 PubMedCrossRefGoogle Scholar
  27. Mohamad O, Yu SP, Chen D, Ogle M, Song M, Wei L (2014) Efficient neuronal differentiation of mouse ES and iPS cells using a rotary cell culture protocol. Differentiation. doi: 10.1016/2013.12.002 Google Scholar
  28. Muth-Kohne E, Pachernegg S, Karus M, Faissner A, Hollmann M (2011) Expression of NMDA receptors and Ca2+-impermeable AMPA receptors requires neuronal differentiation and allows discrimination between two different types of neural stem cells. Cell Physiol Biochem 26(6):935–946. doi: 10.1159/000324002 CrossRefGoogle Scholar
  29. O’Donnell M, Chance RK, Bashaw GJ (2009) Axon growth and guidance: receptor regulation and signal transduction. Annu Rev Neurosci 32:383–412. doi: 10.1146/051508.135614 PubMedCrossRefGoogle Scholar
  30. Ouyang M, Shen X (2006) Critical role of ASK1 in the 6-hydroxydopamine-induced apoptosis in human neuroblastoma SH-SY5Y cells. J Neurochem 97(1):234–244. doi: 10.1111/1471-4159.2006.03730 PubMedCrossRefGoogle Scholar
  31. Pan J, Xiao Q, Sheng CY, Hong Z, Yang HQ, Wang G, Ding JQ, Chen SD (2009) Blockade of the translocation and activation of c-Jun N-terminal kinase 3 (JNK3) attenuates dopaminergic neuronal damage in mouse model of Parkinson’s disease. Neurochem Int 54(7):418–425. doi: 10.1016/2009.01.013 PubMedCrossRefGoogle Scholar
  32. Park G, Yoon BS, Moon JH, Kim B, Jun EK, Oh S, Kim H, Song HJ, Noh JY, Oh C, You S (2008) Green tea polyphenol epigallocatechin-3-gallate suppresses collagen production and proliferation in keloid fibroblasts via inhibition of the STAT3-signaling pathway. J Invest Dermatol 128(10):2429–2441. doi: 10.1038/2008.103 PubMedCrossRefGoogle Scholar
  33. Pool M, Thiemann J, Bar-Or A, Fournier AE (2008) NeuriteTracer: a novel ImageJ plugin for automated quantification of neurite outgrowth. J Neurosci Methods 168(1):134–139. doi: 10.1016/2007.08.029 PubMedCrossRefGoogle Scholar
  34. Qu C, Li W, Shao Q, Dwyer T, Huang H, Yang T, Liu G (2013) c-Jun N-terminal kinase 1 (JNK1) is required for coordination of netrin signaling in axon guidance. J Biol Chem 288(3):1883–1895. doi: 10.1074/112.417881 PubMedCentralPubMedCrossRefGoogle Scholar
  35. Rallis A, Moore C, Ng J (2010) Signal strength and signal duration define two distinct aspects of JNK-regulated axon stability. Dev Biol 339(1):65–77. doi: 10.1016/2009.12.016 PubMedCentralPubMedCrossRefGoogle Scholar
  36. Repici M, Mare L, Colombo A, Ploia C, Sclip A, Bonny C, Nicod P, Salmona M, Borsello T (2009) c-Jun N-terminal kinase binding domain-dependent phosphorylation of mitogen-activated protein kinase kinase 4 and mitogen-activated protein kinase kinase 7 and balancing cross-talk between c-Jun N-terminal kinase and extracellular signal-regulated kinase pathways in cortical neurons. Neuroscience 159(1):94–103. doi: 10.1016/2008.11.049 PubMedCrossRefGoogle Scholar
  37. Ribas VT, Goncalves BS, Linden R, Chiarini LB (2012) Activation of c-Jun N-terminal kinase (JNK) during mitosis in retinal progenitor cells. PLoS ONE 7(4):e34483. doi: 10.1371/0034483 PubMedCentralPubMedCrossRefGoogle Scholar
  38. Song AH, Wang D, Chen G, Li Y, Luo J, Duan S, Poo MM (2009) A selective filter for cytoplasmic transport at the axon initial segment. Cell 136(6):1148–1160. doi: 10.1016/2009.01.016 PubMedCrossRefGoogle Scholar
  39. Theus MH, Wei L, Cui L, Francis K, Hu X, Keogh C, Yu SP (2008) In vitro hypoxic preconditioning of embryonic stem cells as a strategy of promoting cell survival and functional benefits after transplantation into the ischemic rat brain. Exp Neurol 210(2):656–670. doi: 10.1016/2007.12.020 PubMedCrossRefGoogle Scholar
  40. Tiwari VK, Stadler MB, Wirbelauer C, Paro R, Schubeler D, Beisel C (2012) A chromatin-modifying function of JNK during stem cell differentiation. Nat Genet 44(1):94–100. doi: 10.1038/1036 CrossRefGoogle Scholar
  41. Tonges L, Planchamp V, Koch JC, Herdegen T, Bahr M, Lingor P (2011) JNK isoforms differentially regulate neurite growth and regeneration in dopaminergic neurons in vitro. J Mol Neurosci 45(2):284–293. doi: 10.1007/s12031-011-9519-1 PubMedCentralPubMedCrossRefGoogle Scholar
  42. van Inzen WG, Peppelenbosch MP, van den Brand MW, Tertoolen LG, de Laat SW (1996) Neuronal differentiation of embryonic stem cells. Biochim Biophys Acta 1312(1):21–26PubMedCrossRefGoogle Scholar
  43. Waetzig V, Herdegen T (2003) The concerted signaling of ERK1/2 and JNKs is essential for PC12 cell neuritogenesis and converges at the level of target proteins. Mol Cell Neurosci 24(1):238–249PubMedCrossRefGoogle Scholar
  44. Wei L, Cui L, Snider BJ, Rivkin M, Yu SS, Lee CS, Adams LD, Gottlieb DI, Johnson EM Jr, Yu SP, Choi DW (2005) Transplantation of embryonic stem cells overexpressing Bcl-2 promotes functional recovery after transient cerebral ischemia. Neurobiol Dis 19(1–2):183–193. doi: 10.1016/2004.12.016 PubMedCrossRefGoogle Scholar
  45. Weston CR, Davis RJ (2007) The JNK signal transduction pathway. Curr Opin Cell Biol 19(2):142–149. doi: 10.1016/2007.02.001 PubMedCrossRefGoogle Scholar
  46. Wu YY, Bradshaw RA (1996) Induction of neurite outgrowth by interleukin-6 is accompanied by activation of Stat3 signaling pathway in a variant PC12 cell (E2) line. J Biol Chem 271(22):13023–13032PubMedCrossRefGoogle Scholar
  47. Xiao J, Liu Y (2003) Differential roles of ERK and JNK in early and late stages of neuritogenesis: a study in a novel PC12 model system. J Neurochem 86(6):1516–1523PubMedCrossRefGoogle Scholar
  48. Yao K, Ki MO, Chen H, Cho YY, Kim SH, Yu DH, Lee SY, Lee KY, Bae K, Peng C, Lim-do Y, Bode AM, Dong Z (2014) JNK1 and 2 play a negative role in reprogramming to pluripotent stem cells by suppressing Klf4 activity. Stem Cell Res 12(1):139–152. doi: 10.1016/2013.10.005 PubMedCrossRefGoogle Scholar
  49. Yu YM, Han PL, Lee JK (2003) JNK pathway is required for retinoic acid-induced neurite outgrowth of human neuroblastoma, SH-SY5Y. NeuroReport 14(7):941–945. doi: 10.1097/0000074341.81633.b8 PubMedGoogle Scholar
  50. Yu SP, Wei Z, Wei L (2013) Preconditioning strategy in stem cell transplantation therapy. Transl Stroke Res 4(1):76–88. doi: 10.1007/s12975-012-0251-0 PubMedCentralPubMedCrossRefGoogle Scholar
  51. Zhou X, Song M, Chen D, Wei L, Yu SP (2011) Potential role of KCNQ/M-channels in regulating neuronal differentiation in mouse hippocampal and embryonic stem cell-derived neuronal cultures. Exp Neurol. doi: 10.1016/2011.03.018 PubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Zheng Zachory Wei
    • 1
    • 2
  • Shan Ping Yu
    • 1
    • 2
  • Jin Hwan Lee
    • 1
  • Dongdong Chen
    • 1
    • 2
  • Tammi M. Taylor
    • 1
  • Todd Carter Deveau
    • 1
  • Albert Cheung Hoi Yu
    • 3
  • Ling Wei
    • 1
    • 4
    Email author
  1. 1.Department of AnesthesiologyEmory University School of MedicineAtlantaUSA
  2. 2.Center for Visual and Neurocognitive RehabilitationAtlanta VA Medical CenterDecaturUSA
  3. 3.Neuroscience Research Institute and Department of NeurobiologyPeking University School of Basic Medical SciencesBeijingChina
  4. 4.Department of NeurologyEmory University School of MedicineAtlantaUSA

Personalised recommendations