Molecular and Cellular Biochemistry

, Volume 381, Issue 1–2, pp 217–220 | Cite as

The effect of partial outlet obstruction on calpain and phospholipase-2 activities: analyzed by severity and duration

  • Connor M. Callaghan
  • Arnold Johnson
  • Paul Neumann
  • Robert E. Leggett
  • Catherine Schuler
  • Robert M. Levin


In an attempt to better understand the two pathways that lead to bladder decompensation following partial obstruction in rabbits one of which is caused by calcium-activated enzymes and the other by oxidative stress, calpain and phospholipase A2 (PLA2) biochemical assays were conducted to see how bladder decompensation is mediated by these two calcium-activated enzymes. Partial outlet obstructions of varying durations (4, 8, and 12 weeks plus controls) were performed on 32 New Zealand white rabbits. The rabbits were also grouped by severity: control, mild, intermediate, and severe. The activities of Calpain and PLA2 on the muscle tissue of the bladders were analyzed. A stronger correlation was seen between activities and severities as opposed to between activities and durations for both PLA2 and calpain. The activity for PLA2 increased dramatically from control to mild and then stayed constant for both intermediate and severe obstructions. Calpain activity increased steadily from control to mild to intermediate to severe. Based on the increase in levels of the calcium-dependent enzymes, it was clearly shown that calcium levels increased in all stages of bladder decompensation most notably with the mild obstructions. Based on previous studies in which nitrotyrosine and dinitrophenol levels did not increase in mildly obstructed rabbits, the calcium overload pathway may predominate in mild decompensation because cells in mildly obstructed bladders are better able to cope with oxidative stress than increased calcium levels.


Calpain PLA2 Rabbit Urinary bladder Calcium Oxidative stress 


  1. 1.
    Valentini FA, Levin RM, Besson GR, Nelson PP (2003) Study of detrusor dysfunction due to outlet obstruction: link between analysis of uroflows of men with benign prostatic hyperplasia and animal studies. Adv Exp Med Biol 539:297–309PubMedGoogle Scholar
  2. 2.
    Levin RM, O’Connor LJ, Leggett RE, Whitbeck C, Chichester P (2003) Focal hypoxia of the obstructed rabbit bladder wall correlates with intermediate decompensation. Neurourol Urodyn 22:156–163. doi:10.1002/nau.10076 PubMedCrossRefGoogle Scholar
  3. 3.
    Vanarsdalen KN, Wein AJ, Levin RM (1983) The contractile and metabolic effects of acute ischemia on the rabbit urinary bladder. J Urol 130:180–182PubMedGoogle Scholar
  4. 4.
    Gill HS, Monson FC, Wein AJ, Ruggieri MR, Levin RM (1988) The effects of short-term in vivo ischemia on the contractile function of the rabbit urinary bladder. J Urol 139:1350–1354PubMedGoogle Scholar
  5. 5.
    Azadzoi KM, Yalla SV, Siroky MB (2007) Oxidative stress and neurodegeneration in the ischemic overactive bladder. J Urol 178:710–715. doi:10.1016/j.juro.2007.03.096 S0022-5347(07)00744-6 [pii]PubMedCrossRefGoogle Scholar
  6. 6.
    Zhao Y, Levin SS, Wein AJ, Levin RM (1997) Correlation of ischemia/reperfusion or partial outlet obstruction-induced spectrin proteolysis by calpain with contractile dysfunction in rabbit bladder. Urology 49:293–300. doi:10.1016/S0090-4295(96)00452-9 S0090-4295(96)00452-9 [pii]PubMedCrossRefGoogle Scholar
  7. 7.
    Radu F, Leggett RE, Schuler C, Levin RM (2011) The effect of antioxidants on the response of the rabbit urinary bladder to in vitro ischemia/reperfusion. Mol Cell Biochem 355:65–73. doi:10.1007/s11010-011-0839-9 PubMedCrossRefGoogle Scholar
  8. 8.
    Hass MA, Levin RM (2003) The role of lipids and lipid metabolites in urinary bladder dysfunction induced by partial outlet obstruction. Adv Exp Med Biol 539:217–237PubMedGoogle Scholar
  9. 9.
    Levin RM, Schuler C, Leggett RE, Callaghan C, Maknuru S (2012) Partial outlet obstruction in rabbits: duration versus severity. Int J Urol. doi:10.1111/j.1442-2042.2012.03184.x PubMedGoogle Scholar
  10. 10.
    Smith MA, Schnellmann RG (2012) Calpains, mitochondria, and apoptosis. Cardiovasc Res 96:32–37. doi:10.1093/cvr/cvs163 PubMedCrossRefGoogle Scholar
  11. 11.
    Wu J, Liu T, Xie J, Xin F, Guo L (2006) Mitochondria and calpains mediate caspase-dependent apoptosis induced by doxycycline in HeLa cells. Cell Mol Life Sci 63:949–957. doi:10.1007/s00018-005-5565-6 PubMedCrossRefGoogle Scholar
  12. 12.
    Sun GY, Shelat PB, Jensen MB, He Y, Sun AY, Simonyi A (2010) Phospholipases A2 and inflammatory responses in the central nervous system. Neuromolecular Med 12:133–148. doi:10.1007/s12017-009-8092-z PubMedCrossRefGoogle Scholar
  13. 13.
    Tubaro A, Mariani S, De Nunzio C, Miano R (2010) Bladder weight and detrusor thickness as parameters of progression of benign prostatic hyperplasia. Curr Opin Urol 20:37–42. doi:10.1097/MOU.0b013e32833307e0 PubMedCrossRefGoogle Scholar
  14. 14.
    Chacko S, DiSanto M, Wang Z, Zderic SA, Wein AJ (1997) Contractile protein changes in urinary bladder smooth muscle during obstruction-induced hypertrophy. Scand J Urol Nephrol Suppl 184:67–76PubMedGoogle Scholar
  15. 15.
    Levin RM, Schuler C, Leggett RE, Callaghan C, Maknuru S (2013) Partial outlet obstruction in rabbits: duration versus severity. Int J Urol 20:107–114. doi:10.1111/j.1442-2042.2012.03184.x PubMedCrossRefGoogle Scholar
  16. 16.
    Radu F, Leggett RE, Schuler C, Levin RM (2011) The effect of in vitro ischemia/reperfusion on contraction, free fatty acid content, phospholipid content, and malondialdehyde levels of the rabbit urinary bladder. Mol Cell Biochem 346:179–186. doi:10.1007/s11010-010-0603-6 PubMedCrossRefGoogle Scholar
  17. 17.
    Fitzpatrick B, Schuler C, Leggett RE, Levin RM (2012) Calcium effects on superoxide dismutase and catalase of the rabbit urinary bladder muscle and mucosa. Urol Int 88:107–111. doi:10.1159/000333144 000333144 [pii]PubMedCrossRefGoogle Scholar
  18. 18.
    Fitzpatrick B, Schuler C, Leggett RE, Levin RM (2012) Nitrotyrosine Density of Rabbit Urinary Bladder Muscle and Mucosa Measured via Western Blotting and 96-Well Plate Analysis. ISRN Urol 2012:618247. doi:10.5402/2012/618247 PubMedGoogle Scholar
  19. 19.
    Muralikrishna Adibhatla R, Hatcher JF (2006) Phospholipase A2, reactive oxygen species, and lipid peroxidation in cerebral ischemia. Free Radic Biol Med 40:376–387. doi:10.1016/j.freeradbiomed.2005.08.044 PubMedCrossRefGoogle Scholar
  20. 20.
    Kudo I (2004) Diversity of phospholipase A2 enzymes. Foreword. Biol Pharm Bull 27:1157CrossRefGoogle Scholar
  21. 21.
    Kudo I, Murakami M (2002) Phospholipase A2 enzymes. Prostaglandins Other Lipid Mediat 68–69:3–58PubMedCrossRefGoogle Scholar
  22. 22.
    Stefansson BV, Haraldsson B, Nilsson U (2011) Acute oxidative stress following intravenous iron injection in patients on chronic hemodialysis: a comparison of iron-sucrose and iron-dextran. Nephron Clin Pract 118:c249–c256. doi:10.1159/000321645 PubMedCrossRefGoogle Scholar
  23. 23.
    Gupta A, Zhuo J, Zha J, Reddy S, Olp J, Pai A (2010) Effect of different intravenous iron preparations on lymphocyte intracellular reactive oxygen species generation and subpopulation survival. BMC Nephrol 11:16. doi:10.1186/1471-2369-11-16 PubMedCrossRefGoogle Scholar
  24. 24.
    Croall DE, DeMartino GN (1991) Calcium-activated neutral protease (calpain) system: structure, function, and regulation. Physiol Rev 71:813–847PubMedGoogle Scholar
  25. 25.
    Mannikarottu AS, Kogan B, Levin RM (2005) Ischemic etiology of obstructive bladder dysfunction: a review. Recent Res Devel Mol Cell Biochem 2:15–34Google Scholar
  26. 26.
    Buttyan R, Chen MW, Levin RM (1997) Animal models of bladder outlet obstruction and molecular insights into the basis for the development of bladder dysfunction. Eur Urol 32(Suppl 1):32–39PubMedGoogle Scholar
  27. 27.
    Levin RM, Brading AF, Mills IW, Longhust PA (1999) Experimental models of bladder obstruction. In: Lepor H (ed) Prostatic Disease. W.B. Saunders Co, Philadelphia, pp 169–196Google Scholar
  28. 28.
    Hass MA, Leonova E, Levin RM (1999) Fatty acid profiles in normal and obstructed rabbit bladder smooth muscle and mucosa. Neurourol Urodyn 18:697–711PubMedCrossRefGoogle Scholar
  29. 29.
    Nicotera TM, Schuster DP, Bourhim M, Chadha K, Klaich G, Corral DA (2009) Regulation of PSA secretion and survival signaling by calcium-independent phopholipase A(2)beta in prostate cancer cells. Prostate 69:1270–1280. doi:10.1002/pros.20968 PubMedCrossRefGoogle Scholar
  30. 30.
    Lin WY, Wu SB, Lin YP, Chang PJ, Levin RM, Wei YH (2012) Reversing bladder outlet obstruction attenuates systemic and tissue oxidative stress. BJU Int 110:1208–1213. doi:10.1111/j.1464-410X.2012.11185.x PubMedCrossRefGoogle Scholar
  31. 31.
    Bhat MB, Mishra SK, Raviprakash V (1989) Differential susceptibility of cholinergic and noncholinergic neurogenic responses to calcium channel blockers and low Ca2+ medium in rat urinary bladder. Br J Pharmacol 96:837–842PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Connor M. Callaghan
    • 2
  • Arnold Johnson
    • 3
  • Paul Neumann
    • 3
  • Robert E. Leggett
    • 1
  • Catherine Schuler
    • 1
  • Robert M. Levin
    • 1
  1. 1.ResearchStratton VA Medical CenterAlbanyUSA
  2. 2.Pharmacy PracticeAlbany College of Pharmacy and Health SciencesAlbanyUSA
  3. 3.Pharmaceutical SciencesAlbany College of Pharmacy and Health SciencesAlbanyUSA

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