Journal of Molecular Histology

, Volume 43, Issue 2, pp 235–241 | Cite as

Effects of sugammadex on immunoreactivity of calcineurin in rat testes cells after neuromuscular block: a pilot study

  • Yıldıray Kalkan
  • Levent Tümkaya
  • Habib Bostan
  • Yakup Tomak
  • Adnan Yılmaz


Reversal of neuromuscular blockage induced by steroidal neuromuscular blocking agents such as rocuronium can be achieved using normal dose of sugammadex, which has been shown to be very effective for such reversal. In this study, we determined the effects of sugammadex on calcineurin immunoreactivity by examining the histopathological and histochemical structure of rat testis cells after neuromuscular blockage. Moreover, the regional distribution levels of calcineurin immunopositive testes cells were investigated. Eighteen adult male, Sprague–Dawley rats were divided into one control and two study groups. Study groups 1 and 2 rats received sugammadex at doses of 16 and 96 mg kg−1 i.v., respectively, after rocuronium treatment (mg kg−1 i.v.). The control group received intravenous 0.9% NaCl 1 ml. i.v without any drug. Our study demonstrates that sugammadex is safe and effective for reversal of rocuronium effects in rats, as well as in other animals and humans. Furthermore, histopathological examination indicates that high levels of sugammadex–rocuronium complexes accumulate a little in testis tissue. We found that rocuronium-sugammadex complexes were remained in circulation for a long time resulting in a decrease in interstitial space, testis size, germ cell numbers and Leydig cell numbers. Calcineurin immunoreactivity was higher in the experimental groups than the control group due to increase of calcium level. The results suggest that sugammadex–rocuronium complexes are cause histopathological and immunohistochemical changes in testis interstitial tissues, as well as changes in sperm density and germ cell number.


Sugammadex Rocuronium Testis Calcineurin Rat 



The authors declare that they have no conflict of interest. This work was supported by all authors with no external funding from a government agency or company. The animals, materials and drugs were provided by the first author as research fund from his institution (Medical Faculty of Rize University, Department of Histology and Embryology).


  1. Adam JM, Bennett DJ, Bom A, Clark JK, Feilden H, Hutchinson EJ, Palin M, Prosser A, Rees DC, Rosair GM, Stevenson D, Tarver GJ, Zhang MQ (2002) Cyclodextrinderived host molecules as reversal agents for the neuromuscular blocker rocuronium bromide: synthesis and structure-activity relationships. J Med Chem 45:1806–1816. doi: 10.1021/jm011107f PubMedCrossRefGoogle Scholar
  2. Baillard C, Gehan G, Reboul-Marty J, Larmignat P, Samama CM, Cupa M (2000) Residual curarization in the recovery room after vecuronium. Br J Anaesth 84:394–395PubMedCrossRefGoogle Scholar
  3. Baillard C, Clec’h C, Catineau J, Salhi F, Gehan G, Cupa M, Samama CM (2005) Postoperative residual neuromuscular block: a survey of management. Br J Anaesth 95:622–626. doi: 10.1093/bja/aei240 PubMedCrossRefGoogle Scholar
  4. Baldo BA, McDonnell NJ, Pham NH (2011) Drug-specific cyclodextrins with emphasis on sugammadex, the neuromuscular blocker rocuronium and perioperative anaphylaxis: implications for drug allergy. Clin Exp Allergy doi: 10.1111/j.1365-2222.2011.03805.x. [Epub ahead of print]
  5. Bancroft JD, Stevens A (1996) Theory and practice of histological techniques, 4th edn. Churchill-Livingstone, London, pp 42–165Google Scholar
  6. Baumgarten A (1970) Vascular permeability responses in hypersensitivity. 3. Reaction to human immunoglobulin. Pathology 2(1):61–70. doi: 10.3109/00313027009077327 PubMedCrossRefGoogle Scholar
  7. Bergh A, Collin O, Lissbrant E (2001) Effects of acute graded reductions in testicular blood flow on testicular morphology in the adult rat. Biol Reprod 64:13–20. doi: 10.1095/biolreprod64.1.13 PubMedCrossRefGoogle Scholar
  8. Bom A, Bradley M, Cameron K et al (2002) A novel concept of reversing neuromuscular block: chemical encapsulation of rocuronium bromide by a cyclodextrin-based synthetic host. Angew Chem Int Ed Engl 41:266–270PubMedCrossRefGoogle Scholar
  9. Booij LH, Van Egmond J, Driessen JJ, De Boer HD (2009) In vivo animal studies with sugammadex. Anaesthesia 64:38–44. doi: 10.1111/j.1365-2044.2008.05869.x PubMedCrossRefGoogle Scholar
  10. Burburan SM, Xisto DG, Rocco PR (2007) Anaesthetic management in asthma. Minerva Anestesiologica 73(6):357–365PubMedGoogle Scholar
  11. de Boer HD (2009) Neuromuscular transmission: new concepts and agents. J Crit Care 24(1):36–42PubMedCrossRefGoogle Scholar
  12. de Boer HD, van Egmond J, van de Pol F, Bom A, Booij LHDJ (2006) Sugammadex, a new reversal agent for neuromuscular block in anaesthetized Rhesus monkeys. Br J Anaesth 96:473–479. doi: 10.1093/bja/ael013 PubMedCrossRefGoogle Scholar
  13. de Boer HD, van Egmond J, Driessen JJ, Booij LHDJ (2007) Update on the management of neuromuscular block: focus on Sugammadex. Neuropsychiatr Dis Treat 3(5):539–544PubMedGoogle Scholar
  14. Duvaldestin P, Plaud B (2010) Sugammadex in anesthesia practice. Expert Opin Pharmacother 11(16):2759–2771. doi: 10.1517/14656566.2010.528391 PubMedCrossRefGoogle Scholar
  15. Fimmel S, Zouboulis CC (2005) Influence of physiological androgen levels on wound healing and immune status in men. Aging Male 8(3–4):166–174. doi: 10.1080/13685530500233847 PubMedCrossRefGoogle Scholar
  16. Gijsenbergh F, Ramael S, Houwing N, van Iersel T (2005) First human exposure of Org 25969, a novel agent to reverse the action of rocuronium bromide. Anesthesiology 103:695–703. doi: 10.1097/00000542-200510000-00007 PubMedCrossRefGoogle Scholar
  17. Gooch JL (2006) An emerging role for calcineurin Aa in the development and function of the kidney. Am J Physiol Renal Physiol 290:769–776. doi: 10.1152/ajprenal.00281.2005 CrossRefGoogle Scholar
  18. Jones RK, Caldwell JE, Brull SJ, Soto RG (2008) Reversal of profound rocuronium-induced blockade with sugammadex: a randomized comparison with neostigmine. Anesthesiology 109:816–824. doi: 10.1097/ALN.0b013e31818a3fee PubMedCrossRefGoogle Scholar
  19. Klee CB, Draetta GF, Hubbard MJ (1988) Calcineurin. Adv Enzym Relat Areas Mol Biol 61:149–200Google Scholar
  20. Langille BL, O’Donnell F (1986) Reductions in arterial diameter produced by chronic decreases in blood flow are endothelium-dependent. Science 231(4736):405–407. doi: 10.1126/science.3941904 PubMedCrossRefGoogle Scholar
  21. Liu J, Farmer JD Jr, Lane WS, Friedma J, Weissman I, Schreiber SL (1991) Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell 66:807–815. doi: 10.1016/0092-8674(91)90124-H PubMedCrossRefGoogle Scholar
  22. Miyamoto K, Matsui H, Tomizawa K, Kuwata Y, Itano T, Tokuda M, Hatase O (1994) In situ localization of rat testis-specific calcineurin B subunit isoform β1 in the developing rat testis. Biochem Biophys Res Commun 203:1275–1283. doi: 10.1006/bbrc.1994.2320 PubMedCrossRefGoogle Scholar
  23. Moriya M, Fujinaga K, Yazawa M, Katagiri C (1995) Immunohistochemical localization of the calcium/calmodulin-dependent protein phosphatase, calcineurin, in the mouse testis: its unique accumulation in spermatid nuclei. Cell Tissue Res 281(2):273–281. doi: 10.1007/BF00583396 PubMedCrossRefGoogle Scholar
  24. Naguib M (2007) Sugammadex: another milestone in clinical neuromuscular pharmacology. Anesth Analg 104(3):575–581. doi: 10.1213/01.ane.0000244594.63318.fc PubMedCrossRefGoogle Scholar
  25. Nishio H, Matsui K, Tsuji H, Tamura A, Suzuki K (2000) Immunolocalization of calcineurin and FKBP12, the FK506-binding protein, in Hassall’s corpuscles of human thymus and epidermis. Histochem Cell Biol 114(1):9–14PubMedGoogle Scholar
  26. Peeters PA, van den Heuvel MW, van Heumen E, Passier PC, Smeets JM, van Iersel T, Zwiers A (2010) Safety, tolerability and pharmacokinetics of sugammadex using single high doses (up to 96 mg/kg) in healthy adult subjects: a randomized, double-blind, crossover, placebo-controlled, single-centre study. Clin Drug Investig 30(12):867–874. doi: 10.2165/11538920-000000000-00000 PubMedGoogle Scholar
  27. Rusnak F, Mertz P (2000) Calcineurin: form and function. Physiol Rev 80(4):1483–1521PubMedGoogle Scholar
  28. Sorgenfrei IF, Norrild K, Larsen PB, Stensballe J, Østergaard D, Prins ME, Viby-Mogensen J (2006) Reversal of rocuronium-induced neuromuscular block by the selective relaxant binding agent sugammadex: a dose-finding and safety study. Anesthesıology 104:667–674. doi: 10.1097/00000542-200604000-00009 PubMedCrossRefGoogle Scholar
  29. Staals LM, de Boer HD, van Egmond J, Hope F, van de Pol F, Bom AH, Driessen JJ, Booij LH (2011) Reversal of rocuronium-induced neuromuscular block by sugammadex is independent of renal perfusion in anesthetized cats. J Anesth 25(2):241–246PubMedCrossRefGoogle Scholar
  30. Su Q, Zhao M, Weber E, Eugster H-P, Ryffel B (1995) Distribution and activity of calcineurin in rat tissues, evidence for post-transcriptional regulation of testis-specific calcineurin B. Eur J Biochem 230:469–474. doi: 10.1111/j.1432-1033.1995.0469h.x PubMedCrossRefGoogle Scholar
  31. Tash JS, Krinks M, Patel J, Means RL, Klee CB, Means AR (1988) Identification, characterization, and functional correlation of calmodulin-dependent protein phosphatase in sperm. J Cell Biol 106:1625–1633. doi: 10.1083/jcb.106.5.1625 PubMedCrossRefGoogle Scholar
  32. Yamashita M, Katsumata M, Iwashima M, Kimura M, Shimizu C, Kamata T, Shin T, Seki N, Suzuki S, Taniguchi M, Nakayama T (2000) T cell receptor-induced calcineurin activation regulates T helper type 2 cell development by modifying the interleukin 4 receptor signaling complex. J Exp Med 191(11):1869–1879. doi: 10.1084/jem.191.11.1869 PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Yıldıray Kalkan
    • 1
  • Levent Tümkaya
    • 1
  • Habib Bostan
    • 2
  • Yakup Tomak
    • 2
  • Adnan Yılmaz
    • 3
  1. 1.Department of Histology and Embryology, Medical FacultyRize UniversityRizeTurkey
  2. 2.Department of Anesthesiology and Intensive Care, Medical FacultyRize UniversityRizeTurkey
  3. 3.Department of Biochemistry, Medical FacultyRize UniversityRizeTurkey

Personalised recommendations