The Journal of General Physiology
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Published online Jan 31 2005. doi:10.1085/jgp.200409212
The Rockefeller University Press, 0022-1295 $8.00
JGP, Volume 125, Number 2, 187-196
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Decreased Phosphatase Activity, Increased Ca2+ Sensitivity, and Myosin Light Chain Phosphorylation in Urinary Bladder Smooth Muscle of Newborn Mice

Mari Ekman1, Katarina Fagher1, Mia Wede1, Karolina Stakeberg1, and Anders Arner1,2

1 Department of Physiological Sciences, Lund University, S-221 84 Lund, Sweden
2 Department of Physiology and Pharmacology, Karolinska Institute, S-171 77 Stockholm, Sweden

Correspondence to Anders Arner: Anders.Arner{at}fyfa.ki.se

Developmental changes in the regulation of smooth muscle contraction were examined in urinary bladder smooth muscle from mice. Maximal active stress was lower in newborn tissue compared with adult, and it was correlated with a lower content of actin and myosin. Sensitivity to extracellular Ca2+ during high-K+ contraction, was higher in newborn compared with 3-wk-old and adult bladder strips. Concentrations at half maximal tension (EC50) were 0.57 ± 0.01, 1.14 ± 0.12, and 1.31 ± 0.08 mM. Force of the newborn tissue was inhibited by ~45% by the nonmuscle myosin inhibitor Blebbistatin, whereas adult tissue was not affected. The calcium sensitivity in newborn tissue was not affected by Blebbistatin, suggesting that nonmuscle myosin is not a primary cause for increased calcium sensitivity. The relation between intracellular [Ca2+] and force was shifted toward lower [Ca2+] in the newborn bladders. This increased Ca2+ sensitivity was also found in permeabilized muscles (EC50: 6.10 ± 0.07, 5.77 ± 0.08, and 5.55 ± 0.02 pCa units, in newborn, 3-wk-old, and adult tissues). It was associated with an increased myosin light chain phosphorylation and a decreased rate of dephosphorylation. No difference was observed in the myosin light chain phosphorylation rate, whereas the rate of myosin light chain phosphatase–induced relaxation was about twofold slower in the newborn tissue. The decreased rate was associated with a lower expression of the phosphatase regulatory subunit MYPT-1 in newborn tissue. The results show that myosin light chain phosphatase activity can be developmentally regulated in mammalian urinary bladders. The resultant alterations in Ca2+ sensitivity may be of importance for the nervous and myogenic control of the newborn bladders.

Key Words: myosin light chain kinase • myosin light chain phosphatase • MYPT-1 • nonmuscle myosin


Abbreviations used in this paper: ED, embryonic day; LC, light chain; MLCK, myosin light chain kinase; MLCP, myosin light chain phosphatase.


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