Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

S-nitrosylation of NOS pathway mediators in the penis contributes to cavernous nerve injury-induced erectile dysfunction

International Journal of Impotence Research volume 30, pages 108–116 (2018)Cite this article

Subjects

Abstract

cGMP-independent nitric oxide (NO) signaling occurs via S-nitrosylation. We evaluated whether aberrant S-nitrosylation operates in the penis under conditions of cavernous nerve injury and targets proteins involved in regulating erectile function. Adult male Sprague–Dawley rats underwent bilateral cavernous nerve crush injury (BCNI) or sham surgery. Rats were given a denitrosylation agent N-acetylcysteine (NAC, 300 mg/kg/day) or vehicle in drinking water starting 2 days before BCNI and continuing for 2 weeks following surgery. After assessment of erectile function (intracavernous pressure), penes were collected for measurements of S-nitrosylation by Saville–Griess and TMT-switch assays and PKG-I function by immunoblotting of phospho (P)-VASP-Ser-239. Erectile function was decreased (P < 0.05) after BCNI, and it was preserved (P < 0.05) by NAC treatment. Total S-nitrosothiols and total S-nitrosylated proteins were increased (P < 0.05) after BCNI, and these were partially prevented by NAC treatment. S-nitrosylation of sGC was increased (P < 0.05) after BCNI, and it was prevented (P < 0.05) by NAC treatment. S-nitrosylation of eNOS was increased (P < 0.05) after BCNI, and showed a trend towards decrease by NAC treatment. Protein expression of P-VASP-Ser-239 was decreased (P < 0.05) after BCNI, and showed a trend towards increase by NAC treatment. In conclusion, erectile dysfunction following BCNI is mediated in part by S-nitrosylation of eNOS and its downstream signaling mediator GC, while denitrosylation protects erectile function by preserving the NO/cGMP signaling pathway.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Facio F Jr, Burnett AL. Penile rehabilitation and neuromodulation. Sci World J. 2009;9:652–64.

    Article  Google Scholar 

  2. Mulhall JP. Defining and reporting erectile function outcomes after radical prostatectomy: challenges and misconceptions. J Urol. 2009;181:462–71.

    Article  Google Scholar 

  3. Angeloni NL, Bond CW, McVary KT, Podlasek CA. Sonic hedgehog protein is decreased and penile morphology is altered in prostatectomy and diabetic patients. PLoS ONE. 2013;14:e70985.

    Article  Google Scholar 

  4. Iacono F, Giannella R, Somma P, Manno G, Fusco F, Mirone V. Histological alterations in cavernous tissue after radical prostatectomy. J Urol. 2005;173:1673–6.

    Article  Google Scholar 

  5. Hannan JL, Kutlu O, Stopak BL, Liu X, Castiglione F, Hedlund P, et al. Valproic acid prevents penile fibrosis and erectile dysfunction in cavernous nerve-injured rats. J Sex Med. 2014;11:1442–51.

    Article  CAS  Google Scholar 

  6. Albersen M, Fandel TM, Lin G, Wang G, Banie L, Lin CS, Lue TF. Injections of adipose tissue-derived stem cells and stem cell lysate improve recovery of erectile function in a rat model of cavernous nerve injury. J Sex Med. 2010;7:3331–40.

    Article  Google Scholar 

  7. Song KM, Jin HR, Park JM, Choi MJ, Kwon MH, Kwon KD, et al. Intracavernous delivery of stromal vascular fraction restores erectile function through production of angiogenic factors in a mouse model of cavernous nerve injury. J Sex Med. 2014;11:1962–73.

    Article  CAS  Google Scholar 

  8. Li H, Matheu MP, Sun F, Wang L, Sanford MT, Ning H, et al. Low-energy shock wave therapy ameliorates erectile dysfunction in a pelvic neurovascular injuries rat model. J Sex Med. 2016;13:22–32.

    Article  Google Scholar 

  9. Lagoda G, Jin L, Lehrfeld TJ, Liu T, Burnett AL. FK506 and sildenafil promote erectile function recovery after cavernous nerve injury through antioxidative mechanisms. J Sex Med. 2007;4:908–16.

    Article  CAS  Google Scholar 

  10. Lagoda G, Xie Y, Sezen SF, Hurt KJ, Liu L, Musicki B, et al. FK506 neuroprotection after cavernous nerve injury is mediated by thioredoxin and glutathione redox systems. J Sex Med. 2011;8:3325–34.

    Article  CAS  Google Scholar 

  11. Wang H, Ding XG, Li SW, Zheng H, Zheng XM, Navin S, Li L, Wang XH. Role of oxidative stress in surgical cavernous nerve injury in a rat model. J Neurosci Res. 2015;93:922–9.

    Article  CAS  Google Scholar 

  12. Hannan JL, Matsui H, Sopko NA, Liu X, Weyne E, Albersen M, et al. Caspase-3 dependent nitrergic neuronal apoptosis following cavernous nerve injury is mediated via RhoA and ROCK activation in major pelvic ganglion. Sci Rep. 2016;6:29416.

    Article  CAS  Google Scholar 

  13. Lee CH, Kim HS, Goo MJ, Kang KK, Ahn BO, Kim SH, et al. Chronic administration of udenafil, a selective phosphodiesterase type 5 inhibitor, promotes erectile function recovery in an animal model of bilateral cavernous nerve crush injury. J Sex Med. 2011;8:1330–40.

    Article  CAS  Google Scholar 

  14. Shindel AW, Xin ZC, Lin G, Fandel TM, Huang YC, Banie L, et al. Erectogenic and neurotrophic effects of icariin, a purified extract of horny goat weed (Epimedium spp.) in vitro and in vivo. J Sex Med. 2010;7:1518–28.

    Article  CAS  Google Scholar 

  15. Hannan JL, Albersen M, Kutlu O, Gratzke C, Stief CG, Burnett AL, et al. Inhibition of Rho-kinase improves erectile function, increases nitric oxide signaling and decreases penile apoptosis in a rat model of cavernous nerve injury. J Urol. 2013;189:1155–61.

    Article  CAS  Google Scholar 

  16. Aliperti LA, Lasker GF, Hagan SS, Hellstrom JA, Gokce A, Trost LW, et al. Efficacy of pioglitazone on erectile function recovery in a rat model of cavernous nerve injury. Urology. 2014;84:1122–7.

    Article  Google Scholar 

  17. Ignarro LJ, Bush PA, Buga GM, Wood KS, Fukuto JM, Rajfer J. Nitric oxide and cyclic GMP formation upon electrical field stimulation cause relaxation of corpus cavernosum smooth muscle. Biochem Biophys Res Commun. 1990;170:843–50.

    Article  CAS  Google Scholar 

  18. Burnett AL. Nitric oxide in the penis: physiology and pathology. J Urol. 1997;157:320–4.

    Article  CAS  Google Scholar 

  19. Lima B, Forrester MT, Hess DT, Stamler JS. S-nitrosylation in cardiovascular signaling. Circ Res. 2010;106:633–46.

    Article  CAS  Google Scholar 

  20. Fulton DJ, Transcriptional and posttranslational regulation of eNOS in the endothelium. Adv Pharmacol. 2016;77:29–64.

    Article  CAS  Google Scholar 

  21. Anand P, Stamler JS. Enzymatic mechanisms regulating protein S-nitrosylation: implications in health and disease. J Mol Med (Berl). 2012;90:233–44.

    Article  CAS  Google Scholar 

  22. Sun J, Murphy E. Protein S-nitrosylation and cardioprotection. Circ Res. 2010;106:285–96.

    Article  CAS  Google Scholar 

  23. Nakamura T, Lipton SA. Emerging role of protein-protein transnitrosylation in cell signaling pathways. Antioxid Redox Signal. 2013;18:239–49.

    Article  CAS  Google Scholar 

  24. Musicki B, Lagoda G, Goetz T, La Favor JD, Burnett AL. Transnitrosylation: a factor in nitric oxide-mediated penile erection. J Sex Med. 2016;13:808–14.

    Article  Google Scholar 

  25. Burnett AL, Sezen SF, Hoke A, Caggiano AO, Iaci J, Lagoda G, et al. GGF2 is neuroprotective in a rat model of cavernous nerve injury-induced erectile dysfunction. J Sex Med. 2015;12:897–905.

    Article  CAS  Google Scholar 

  26. Fioramonti X, Deak A, Deshpande S, Carneiro L, Zhou C, Sayed N, et al. Hypothalamic S-nitrosylation contributes to the counter-regulatory response impairment following recurrent hypoglycemia. PLoS ONE. 2013;8:e68709.

    Article  CAS  Google Scholar 

  27. Hoffmann J, Haendeler J, Zeiher AM, Dimmeler S. TNFalpha and oxLDL reduce protein S-nitrosylation in endothelial cells. J Biol Chem. 2001;276:41383–7.

    Article  CAS  Google Scholar 

  28. Eu JP, Liu L, Zeng M, Stamler JS. An apoptotic model for nitrosative stress. Biochemistry. 2000;39:1040–7.

    Article  CAS  Google Scholar 

  29. Murray CI, Uhrigshardt H, O’Meally RN, Cole RN, Van Eyk JE. Identification and quantification of S-nitrosylation by cysteine reactive tandem mass tag switch assay. Mol Cell Proteomics. 2012;11:013441. M111

    Article  Google Scholar 

  30. Hurt KJ, Musicki B, Palese MA, Crone JK, Becker RE, Moriarity JL, et al. Akt-dependent phosphorylation of endothelial nitric-oxide synthase mediates penile erection. Proc Natl Acad Sci USA. 2002;99:4061–6.

    Article  CAS  Google Scholar 

  31. Musicki B, Liu T, Lagoda GA, Strong TD, Sezen SF, Johnson JM, et al. Hypercholesterolemia-induced erectile dysfunction: endothelial nitric oxide synthase (eNOS) uncoupling in the mouse penis by NAD(P)H oxidase. J Sex Med. 2010;7:3023–32.

    Article  CAS  Google Scholar 

  32. Ludbrook J. Should we use one-sided or two-sided P values in tests of significance? Clin Exp Pharmacol Physiol. 2013;40:357–61.

    Article  CAS  Google Scholar 

  33. Oelze M, Mollnau H, Hoffmann N, Warnholtz A, Bodenschatz M, Smolenski A, et al. Vasodilator-stimulated phosphoprotein serine 239 phosphorylation as a sensitive monitor of defective nitric oxide/cGMP signaling and endothelial dysfunction. Circ Res. 2000;87:999–1005.

    Article  CAS  Google Scholar 

  34. Erwin PA, Lin AJ, Golan DE, Michel T. Receptor-regulated dynamic S-nitrosylation of endothelial nitric-oxide synthase in vascular endothelial cells. J Biol Chem. 2005;280:19888–94.

    Article  CAS  Google Scholar 

  35. Qu ZW, Miao WY, Hu SQ, Li C, Zhuo XL, Zong YY, et al. N-methyl-D-aspartate receptor- dependent denitrosylation of neuronal nitric oxide synthase increase the enzyme activity. PLoS ONE. 2012;7:e52788.

    Article  CAS  Google Scholar 

  36. Erwin PA, Mitchell DA, Sartoretto J, Marletta MA, Michel T. Subcellular targeting and differential S-nitrosylation of endothelial nitric-oxide synthase. J Biol Chem. 2006;281:151–7.

    Article  CAS  Google Scholar 

  37. Ravi K, Brennan LA, Levic S, Ross PA, Black SM. S-nitrosylation of endothelial nitric oxide synthase is associated with monomerization and decreased enzyme activity. Proc Natl Acad Sci USA. 2004;101:2619–24.

    Article  CAS  Google Scholar 

  38. Mulhall JP, Müller A, Donohue JF, Mullerad M, Kobylarz K, Paduch DA, et al. The functional and structural consequences of cavernous nerve injury are ameliorated by sildenafil citrate. J Sex Med. 2008;5:1126–36.

    Article  CAS  Google Scholar 

  39. Sayed N, Baskaran P, Ma X, van den Akker F, Beuve A. Desensitization of soluble guanylyl cyclase, the NO receptor, by S-nitrosylation. Proc Natl Acad Sci USA. 2007;104:12312–7.

    Article  CAS  Google Scholar 

  40. Sayed N, Kim DD, Fioramonti X, Iwahashi T, Durán WN, Beuve A. Nitroglycerin-induced S-nitrosylation and desensitization of soluble guanylyl cyclase contribute to nitrate tolerance. Circ Res. 2008;103:606–14.

    Article  CAS  Google Scholar 

  41. Beuve A, Wu C, Cui C, Liu T, Jain MR, Huang C, et al. Identification of novel S-nitrosation sites in soluble guanylyl cyclase, the nitric oxide receptor. J Proteom. 2016;138:40–47.

    Article  CAS  Google Scholar 

  42. Rajagopal S, Nalli AD, Kumar DP, Bhattacharya S, Hu W, Mahavadi S, et al. Cytokine-induced S-nitrosylation of soluble guanylyl cyclase and expression of phosphodiesterase 1A contribute to dysfunction of longitudinal smooth muscle relaxation. J Pharmacol Exp Ther. 2015;352:509–18.

    Article  Google Scholar 

  43. Zamorano P, Marín N, Córdova F, Aguilar A, Meininger C, Boric MP, et al. S-nitrosylation of VASP at cysteine 64 mediates the inflammation-stimulated increase in microvascular permeability. Am J Physiol Heart Circ Physiol. 2017;313:H66–H71.

    Article  Google Scholar 

  44. Zafarullah M, Li WQ, Sylvester J, Ahmad M. Molecular mechanisms of N-acetylcysteine actions. Cell Mol Life Sci. 2003;60:6–20.

    Article  CAS  Google Scholar 

  45. Yu HM, Xu J, Li C, Zhou C, Zhang F, Han D, et al. Coupling between neuronal nitric oxide synthase and glutamate receptor 6-mediated c-Jun N-terminal kinase signaling pathway via S-nitrosylation contributes to ischemia neuronal death. Neuroscience. 2008;155:1120–32.

    Article  CAS  Google Scholar 

  46. Palmer LA, Kimberly deRonde, Brown-Steinke K, Gunter S, Jyothikumar V, Forbes MS, et al. Hypoxia-induced changes in protein s-nitrosylation in female mouse brainstem. Am J Respir Cell Mol Biol. 2015;52:37–45.

    Article  Google Scholar 

  47. Liao CH, Wu YN, Chen BH, Lin Y, Ho HO, Chiang HS. Neuroprotective effect of docosahexaenoic acid nanoemulsion on erectile function in a rat model of bilateral cavernous nerve injury. Sci Rep. 2016;6:33040.

    Article  CAS  Google Scholar 

  48. Chen X, Yang Q, Zheng T, Bian J, Sun X, Shi Y, et al. Neurotrophic effect of adipose tissue- derived stem cells on erectile function recovery by pigment epithelium-derived factor secretion in a rat model of cavernous nerve injury. Stem Cells Int. 2016;2016:5161248.

    PubMed  Google Scholar 

  49. Forrester MT, Foster MW, Stamler JS. Assessment and application of the biotin switch technique for examining protein S-nitrosylation under conditions of pharmacologically induced oxidative stress. J Biol Chem. 2007;282:13977–83.

    Article  CAS  Google Scholar 

  50. Nakazawa H, Chang K, Shinozaki S, Yasukawa T, Ishimaru K, Yasuhara S, et al. iNOS as a driver of inflammation and apoptosis in mouse skeletal muscle after burn injury: possible involvement of Sirt1 S-Nitrosylation-mediated acetylation of p65 NF-κB and p53. PLoS ONE. 2017;12:e017039.

    Google Scholar 

Download references

Acknowledgements

This work was supported by a grant from the National Institutes of Health, USA (NIH/NIDDK grant R01DK067223 to ALB).

Author information

Authors and Affiliations

  1. The James Buchanan Brady Urological Institute and Department of Urology, The Johns Hopkins School of Medicine, Baltimore, MD, USA

    Biljana Musicki, Anil K. Bhunia, Serkan Karakus & Arthur L. Burnett

Authors
  1. Biljana Musicki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anil K. Bhunia
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Serkan Karakus
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Arthur L. Burnett
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Biljana Musicki.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Musicki, B., Bhunia, A.K., Karakus, S. et al. S-nitrosylation of NOS pathway mediators in the penis contributes to cavernous nerve injury-induced erectile dysfunction. Int J Impot Res 30, 108–116 (2018). https://doi.org/10.1038/s41443-018-0021-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41443-018-0021-y

This article is cited by

Search

Advanced search

Quick links