Abstract
The urinary bladder depends on intracellular ATP to support a number of essential intracellular processes including contraction. The concentration of ATP is maintained by mitochondrial oxidative phosphorylation, cytosolic glycolysis and the cytosolic activity of creatine kinase, the enzyme that catalysis the rapid transfer of a phosphate from creatine phosphate (CP) to ADP resulting in the formation of ATP.
Prior studies in this lab and others have demonstrated that mitochondrial respiration is significantly lower in hypertrophied bladder tissue (induced by partial outlet obstruction of the white New Zealand Rabbit). In addition to decreased mitochondrial respiration, there are significant increases in glycolysis and lactic acid formation in the hypertrophied tissue.
In view of the increased glycolysis and decreased mitochondrial function in the hypertrophied tissue, and the importance in creatine kinase in maintaining cytosolic levels of ATP, the current study was designed to determine if outlet obstruction induces any changes in the activity of creatine kinase.
The following is a summary of the results: 1) The bladder mass increased from 2.2 ± 0.2 gm to 11.5 ±1.6 gm at 7 days following outlet obstruction. 2) The intracellular concentrations of both ATP and CP were significantly reduced in the bladder tissue following 7 days of obstruction. 3) The percent of protein (per tissue mass) was significantly lower in the obstructed bladders, although the percent of soluble protein was similar. 4) Creatine kinase activity of control bladders showed linear kinetics with a Vmax = 1120 nmoles/mg protein/4 min and Km = 147 µM CP. 2) The creatine kinase activity of obstructed bladders also displayed linear kinetics with a Vmax = 1125 nmoles/mg protein/4 min tissue, and Km = 276 µM CP.
These studies demonstrate that whereas both control and obstructed bladders have virtually identical maximum creatine kinase activities, the Km for the obstructed tissue is significantly higher than the Km for the control tissue. This may indicate that under cellular conditions (at sub-maximum substrate concentrations), the creatine kinase activity of the obstructed bladders may be significantly lower than the activity of the control bladders. In addition, the reduced tissue concentrations of ATP and CP would certainly be consistent with the reduced functional response to bethanechol and field stimulation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arner A, Hellstrand P, Ruegg JC: Influence of ATP, ADP and AMPPNP on the energetics of contraction in skinned smooth muscle. Prog Clin Biol Res 245: 43–57, 1987
Stephens NL, Wrogemann K: Oxidative phosphorylation in smooth muscle. In: NL Stephens (ed.) Biochemistry of smooth muscle. Vol II, CRC Press, Inc., Boca Raton, Florida, 1983, pp 119–126
Scott DP, Coburn RF: Phosphocreatine and oxidative metabolism-contraction coupling in rabbit aorta. Am J Physiology 257: H597–602, 1989
Butler TM, Siegman MJ: Chemical energetics of contraction in mammalian smooth muscle. Fed Proc Feb 41(2): 204–208, 1982
Paul RJ: Smooth muscle energetics. Annu Rev Physiol 51: 331–349, 1989
Malkowicz SB, Wein AJ, Elbadawi A, Van Arsdalen K, Ruggieri MR, Levin RM: Acute biochemical and functional alterations in the partially obstructed rabbit urinary bladder. J Urol 136: 1324–1329, 1986
Uvelius B, Persson L, Mattiasson A: Smooth muscle cell hypertrophy and hyperplasia in the rat detrusor after short-time infravesical outflow obstruction. J Urol 131: 173–176, 1984
Mayo ME, Hinman F: Structure and function of the rabbit bladder altered by chronic obstruction or cystitis. Invest Urol 14: 6–9, 1976
Kato K, Lin AT-L, Wein AJ, Levin RM: Effect of outlet obstruction on glucose metabolism of the rabbit urinary bladder. J Urol 143: 844–847, 1990
Arner A, Malmqvist U, Uvelius B: Metabolism and force in hypertrophic smooth muscle from rat urinary bladder. Am J Physiol 258: C923-C932, 1990
Levin RM, Hypolite J, Ruggieri MR, Longhurst PA, Wein AJ: Effects of muscarinic stimulation on intracellular calcium in the rabbit bladder: Comparison with metabolic response, Pharmacol 39: 69–77, 1989
Levin RM, Longhurst PA, Levin SS, Haugaard N, Wein AJ: Creatine kinase activity of urinary bladder and skeletal muscle isolated from normal and streptozotocin-diabetic rat. Mol Cell Biochem 97: 153–159, 1990
Levin RM, Wein AJ: Response of the in vitro whole bladder (rabbit) preparation to autonomic agonists. J Urology 128: 1087–1090, 1982
Levin RM, Brendler K, Wein AJ: Comparative pharmacological response of an in vitro whole bladder preparation (rabbit) with the response of isolated smooth muscle strips. J Urology 30: 377–381, 1983
Haugaard N, Wein AJ, Levin RM: In vitro studies of glucose metabolism of the rabbit urinary bladder. J Urology 137: 782–784, 1986
Levin RM, Ruggieri MR, Gill HS, Haugaard N, Wein AJ: Effect of bethanechol on glycolysis and high energy phosphate metabolism of the rabbit urinary bladder. J Urology 139: 646–649, 1988
Wendt IR: Effects of substrate and hypoxis on smooth metabolism and contraction. Department of Physiology, Monash University, Clayton, Victoria, Australia. Am J Physiol Apr; 256 (4Pt 1): C719–727, 1989
Wendt IR: Energy expenditure of longitudinal smooth muscle of rabbit urinary bladder. Am J Physiol Jan; 252 (1Pt 1): C88-C96, 1987
Ruggieri MR, Wein AJ, Hypolite JA, Levin RM: Dissociation of the metabolic from contractile response to muscarinic receptor stimulation in the rabbit urinary bladder. Mol Cell Biochem 81: 137–143, 1988
Mattiasson A, Uvelius B: Changes in contractile properties in hypertrophic rat urinary bladder. J Urol 128: 1340–1342, 1982
Brent L, Stephens FD: The response of smooth muscle cells in the rabbit urinary bladder to outflow obstruction. Invest Urol 12: 494–502, 1975
Mostwin JL, Brooks L: A new guinea pig model of urethral obstruction. J Urol 141: 335A, 1989
Kato K, Wein AJ, Kitada S, Haugaard N, Levin RM: The functional effect of mild outlet obstruction on the rabbit urinary bladder. J Urol 140: 880–884, 1988
Kim Y, Samuel M, Levin RM, Chacko S: Alterations of proteins associated with cytoplasmic filaments in the hypertrophied urinary bladder. J Urol (in press)
Uvelius B, Arner A, Malmquist U: Contractile and cytoskeletal proteins in detrusor muscle from obstructed rat and human bladder. Neurourol Urodyn 8: 396–398, 1989
Kato K, Wein AJ, Radzinski C, Longhurst PA, McGuire EJ, Miller LF, Elbadawi A, Levin RM: Short term functional effects of bladder outlet obstruction in the cat. J Urol 143: 1020–1025, 1990
Campbell JD, Agubosim S, Paul RJ: Compartmentation of metabolism and function in vascular smooth muscle: Quantitation of Na-pump activity and aerobic glycolysis. FASEB J 2(4): A755, 1988
Ishida Y, Paul RJ: Evidence for compartmentation of highenergy phosphagens in smooth muscle. In: RJ Paul, G Elzinga, K Yamada (eds.) Muscle Energetics, Alan R Liss, NY, 1989, pp 417–429
Lynch RM, Paul RJ: Functional compartmentation of carbohydrate metabolism. In: Dean P Jones (ed.) Microcompartmentation. CRC Crit Rev, Boca Raton, 1989, pp 17–36
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Levin, R.M., Haugaard, N., Levin, S.S. et al. Creatine kinase activity in normal and hypertrophied rabbit urinary bladder tissue (following partial outlet obstruction). Mol Cell Biochem 106, 143–149 (1991). https://doi.org/10.1007/BF00230180
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00230180