Advertisement

Journal of Molecular Neuroscience

, Volume 54, Issue 3, pp 360–369 | Cite as

Role of c-Jun N-Terminal Kinase (JNK) Activation in Micturition Reflexes in Cyclophosphamide (CYP)-Induced Cystitis in Female Rats

  • C. Dugan
  • S. Malley
  • L. Arms
  • V. May
  • M. A. Vizzard
Article

Abstract

c-Jun N-terminal kinase (JNK) is member of the mitogen-activated protein kinase (MAPK) family, activated through phosphorylation following cytokine exposure and stress. In this study, phosphorylation of JNK was examined in the urinary bladder with cyclophosphamide (CYP)-induced cystitis and the effects of SP600125, a selective inhibitor of phosphorylation of JNK, on urinary bladder function were assessed using conscious, open outlet, cystometry with continuous instillation of intravesical saline. We induced bladder inflammation in adult female Wistar rats by injecting CYP intraperitoneally to produce acute (150 mg/kg; 4 h), intermediate (150 mg/kg; 48 h), and chronic (75 mg/kg; every third day for 10 days) treatments. Western blotting of urinary bladder demonstrated a significant (p ≤ 0.01) increase (i.e., phosphorylation) in JNK activation with 4- and 48-h CYP-induced cystitis. Immunohistochemistry and image analyses demonstrated a significant (p ≤ 0.01) increase in JNK activation in the urothelium with 4- and 48-h CYP-induced cystitis. Blockade of JNK phosphorylation significantly (p ≤ 0.01) increased bladder capacity and intercontraction void intervals in CYP-treated rats (4 and 48 h). Furthermore, blockade of JNK phosphorylation reduced (p ≤ 0.01) neuropeptide (substance P, calcitonin gene-related peptide) expression in the urinary bladder with CYP-induced cystitis (4 and 48 h). In contrast, blockade of JNK phosphorylation was without effect on bladder function or neuropeptide expression in urinary bladder in control (no inflammation) rats. Blockade of JNK phosphorylation may represent a novel target for improving urinary bladder function with CYP-induced cystitis.

Keywords

Micturition Western blot Cystometry Phosphorylation IHC Neuropeptides 

Notes

Acknowledgments

This work was funded by National Institutes of Health (NIH) grants DK051369 (MAV), DK060481 (MAV) and DK065989 (MAV). This publication was also supported by grants from the National Center for Research Resources (5 P30 RR 032135) and the National Institute of General Medical Sciences (8 P30 GM 103498) from the NIH. The authors gratefully acknowledge the technical expertise and support provided by the VT Cancer Center DNA Analysis Facility.

References

  1. Arms L, Girard BM, Vizzard MA (2010) Expression and function of CXCL12/CXCR4 in rat urinary bladder with cyclophosphamide-induced cystitis. Am J Physiol Ren Physiol 298:F589–F600CrossRefGoogle Scholar
  2. Arms L, Girard BM, Malley SE, Vizzard MA (2013) Expression and function of CCL2/CCR2 in rat micturition reflexes and somatic sensitivity with urinary bladder inflammation. Am J Physiol Ren Physiol 305:F111–F122CrossRefGoogle Scholar
  3. Bennett BL, Sasaki DT, Murray BW, O’Leary EC, Sakata ST, Xu W, Leisten JC et al (2001) SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc Natl Acad Sci U S A 98:13681–13686PubMedCentralPubMedCrossRefGoogle Scholar
  4. Chung CW, Zhang QL, Qiao LY (2010) Endogenous nerve growth factor regulates collagen expression and bladder hypertrophy through Akt and MAPK pathways during cystitis. J Biol Chem 285:4206–4212PubMedCentralPubMedCrossRefGoogle Scholar
  5. Corrow KA, Vizzard MA (2007) Phosphorylation of extracellular signal-regulated kinases in urinary bladder in rats with cyclophosphamide-induced cystitis. Am J Physiol Regul Integr Comp Physiol 293:R125–R134PubMedCrossRefGoogle Scholar
  6. Corrow KA, Vizzard MA (2009) Phosphorylation of extracellular signal-regulated kinases in bladder afferent pathways with cyclophosphamide-induced cystitis. Neuroscience 163:1353–1362PubMedCentralPubMedCrossRefGoogle Scholar
  7. Corrow K, Girard BM, Vizzard MA (2010) Expression and response of acid-sensing ion channels in urinary bladder to cyclophosphamide-induced cystitis. Am J Physiol Ren Physiol 298:F1130–F1139CrossRefGoogle Scholar
  8. Daulhac L, Mallet C, Courteix C, Etienne M, Duroux E, Privat AM, Eschalier A et al (2006) Diabetes-induced mechanical hyperalgesia involves spinal mitogen-activated protein kinase activation in neurons and microglia via N-methyl-d-aspartate-dependent mechanisms. Mol Pharmacol 70:1246–1254PubMedCrossRefGoogle Scholar
  9. Davis RJ (2000) Signal transduction by the JNK group of MAP kinases. Cell 103:239–252PubMedCrossRefGoogle Scholar
  10. de Launay D, van de Sande MG, de Hair MJ, Grabiec AM, van de Sande GP, Lehmann KA, Wijbrandts CA et al (2012) Selective involvement of ERK and JNK mitogen-activated protein kinases in early rheumatoid arthritis (1987 ACR criteria compared to 2010 ACR/EULAR criteria): a prospective study aimed at identification of diagnostic and prognostic biomarkers as well as therapeutic targets. Ann Rheum Dis 71:415–423PubMedCentralPubMedCrossRefGoogle Scholar
  11. Doya H, Ohtori S, Fujitani M, Saito T, Hata K, Ino H, Takahashi K et al (2005) c-Jun N-terminal kinase activation in dorsal root ganglion contributes to pain hypersensitivity. Biochem Biophys Res Comm 335:132–138PubMedCrossRefGoogle Scholar
  12. Driscoll A, Teichman JM (2001) How do patients with interstitial cystitis present? J Urol 166:2118–2120PubMedCrossRefGoogle Scholar
  13. Gao YJ, Xu ZZ, Liu YC, Wen YR, Decosterd I, Ji RR (2010) The c-Jun N-terminal kinase 1 (JNK1) in spinal astrocytes is required for the maintenance of bilateral mechanical allodynia under a persistent inflammatory pain condition. Pain 148:309–319PubMedCentralPubMedCrossRefGoogle Scholar
  14. Gonzalez EJ, Girard BM, Vizzard MA (2013) Expression and function of transforming growth factor-beta isoforms and cognate receptors in the rat urinary bladder following cyclophosphamide-induced cystitis. Am J Physiol Ren Physiol 305:F1265–F1276CrossRefGoogle Scholar
  15. Hanno PM, Sant GR (2001) Clinical highlights of the National Institute of Diabetes and Digestive and Kidney Diseases/Interstitial Cystitis Association scientific conference on interstitial cystitis. Urology 57:2–6PubMedCrossRefGoogle Scholar
  16. Ikeda H, Kiritoshi T, Murase K (2012) Contribution of microglia and astrocytes to the central sensitization, inflammatory and neuropathic pain in the juvenile rat. Mol Pain 8:43PubMedCentralPubMedCrossRefGoogle Scholar
  17. Lee JY, Choi DC, Oh TH, Yune TY (2013) Analgesic effect of acupuncture is mediated via inhibition of JNK activation in astrocytes after spinal cord injury. PLoS One 8:e73948PubMedCentralPubMedCrossRefGoogle Scholar
  18. Lei L, Yuan X, Wang S, Zhang F, Han Y, Ning Q, Luo G et al (2012) Mitogen-activated protein kinase pathways are involved in the upregulation of calcitonin gene-related peptide of rat trigeminal ganglion after organ culture. J Mol Neurosci 48:53–65PubMedCrossRefGoogle Scholar
  19. Markevich NI, Hoek JB, Kholodenko BN (2004) Signaling switches and bistability arising from multisite phosphorylation in protein kinase cascades. J Cell Biol 164:353–359PubMedCentralPubMedCrossRefGoogle Scholar
  20. Mehan S, Meena H, Sharma D, Sankhla R (2011) JNK: a stress-activated protein kinase therapeutic strategies and involvement in Alzheimer’s and various neurodegenerative abnormalities. J Mol Neurosci 43:376–390PubMedCrossRefGoogle Scholar
  21. Mercer BA, D’Armiento JM (2006) Emerging role of MAP kinase pathways as therapeutic targets in COPD. Intl J Chron Obstructive Pulm Dis 1:137–150Google Scholar
  22. Merrill L, Girard B, Arms L, Guertin P, Vizzard MA (2013a) Neuropeptide/receptor expression and plasticity in micturition pathways. Curr Pharm Des 19:4411–4422PubMedCrossRefGoogle Scholar
  23. Merrill L, Malley S, Vizzard MA (2013b) Repeated variate stress in male rats induces increased voiding frequency, somatic sensitivity, and urinary bladder nerve growth factor expression. Am J Physiol Regul Integr Comp Physiol 305:R147–R156PubMedCentralPubMedCrossRefGoogle Scholar
  24. Obata K, Yamanaka H, Kobayashi K, Dai Y, Mizushima T, Katsura H, Fukuoka T et al (2004) Role of mitogen-activated protein kinase activation in injured and intact primary afferent neurons for mechanical and heat hypersensitivity after spinal nerve ligation. J Neurosci 24:10211–10222PubMedCrossRefGoogle Scholar
  25. Peti W, Page R (2013) Molecular basis of MAP kinase regulation. Protein Sci 22:1698–1710PubMedCrossRefGoogle Scholar
  26. Quillin RB, Erickson DR (2012) Management of interstitial cystitis/bladder pain syndrome: a urology perspective. Urol Clin N Am 39:389–396CrossRefGoogle Scholar
  27. Sant GR, Hanno PM (2001) Interstitial cystitis: current issues and controversies in diagnosis. Urology 57:82–88PubMedCrossRefGoogle Scholar
  28. Schnegelsberg B, Sun TT, Cain G, Bhattacharya A, Nunn PA, Ford AP, Vizzard MA et al (2010) Overexpression of NGF in mouse urothelium leads to neuronal hyperinnervation, pelvic sensitivity, and changes in urinary bladder function. Am J Physiol Regul Integr Comp Physiol 298:R534–R547PubMedCentralPubMedCrossRefGoogle Scholar
  29. Vizzard MA (2000) Changes in urinary bladder neurotrophic factor mRNA and NGF protein following urinary bladder dysfunction. Exp Neurol 161:273–284PubMedCrossRefGoogle Scholar
  30. Vizzard MA (2001) Alterations in neuropeptide expression in lumbosacral bladder pathways following chronic cystitis. J Chem Neuroanat 21:125–138PubMedCrossRefGoogle Scholar
  31. Wang S, Zhang C, Sheng X, Zhang X, Wang B, Zhang G (2014) Peripheral expression of MAPK pathways in Alzheimer’s and Parkinson’s diseases. J Clin Neurosci 21(5):709–892CrossRefGoogle Scholar
  32. Zhuang ZY, Wen YR, Zhang DR, Borsello T, Bonny C, Strichartz GR, Decosterd I et al (2006) A peptide c-Jun N-terminal kinase (JNK) inhibitor blocks mechanical allodynia after spinal nerve ligation: respective roles of JNK activation in primary sensory neurons and spinal astrocytes for neuropathic pain development and maintenance. J Neurosci 26:3551–3560PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • C. Dugan
    • 1
  • S. Malley
    • 1
  • L. Arms
    • 1
  • V. May
    • 1
  • M. A. Vizzard
    • 1
  1. 1.Department of Neurological SciencesUniversity of Vermont College of MedicineBurlingtonUSA

Personalised recommendations