Carbachol-induced sustained tonic contraction of rat detrusor muscle.
OBJECTIVE
To investigate the underlying contractile mechanism of the sustained tonic contraction (SuTC) induced by repetitive carbachol application in rat detrusor muscles.


METHODS
Longitudinal muscle strips with no mucosa were obtained from the anterior wall of the urinary bladder in 12-week-old Sprague-Dawley rats. Carbachol (5 micromol/L) was applied repetitively to induce SuTC. The carbachol-induced SuTC was assessed in the presence of various Ca2+-channel blockers and drugs affecting intracellular Ca2+ concentration.


RESULTS
The first application of carbachol elicited a large phasic contraction followed by a tonic contraction (TC); the carbachol-induced contraction was completely reversed by washing out the solution. However, the initial phasic contraction was not reproduced after a second or further application of carbachol. There was consistently only a SuTC with no phasic contraction. The amplitude of the SuTC was 85% of the TC induced by the first carbachol application. The application of atropine (1 micromol/L) to the bath completely blocked SuTC. The carbachol-induced SuTC was insensitive to nicardipine (5 micromol/L) and extracellular polyvalent cations (1 mmol/L, La3+, Co2+, Cd2+, Ni2+ ). Moreover, a similar SuTC was induced even after the complete elimination of extracellular Ca2+ by adding 2 mmol/L EGTA to the Ca2+-free Tyrode solution. To exclude intracellular Ca2+ sources related to the sarcoplasmic reticulum (SR), the effects of SR Ca2+ pump inhibitors, cyclopiazonic acid (CPA, 10 micromol/L) and thapsigargin (0.5 micromol/L) were tested. The carbachol-induced SuTC was insensitive to pretreatment with CPA and/or thapsigargin. To deplete the ryanodine-sensitive Ca2+ pool, muscle strips were repetitively stimulated with caffeine (10 mmol/L) in the presence of 10 micromol/L ryanodine, which did not affect the carbachol-induced SuTC.


CONCLUSIONS
Although the characteristics of the carbachol-induced SuTC have not been defined, these results show that a significant proportion of the carbachol-induced contraction in rats is contributed by the SuTC, which is present even in the complete absence of external Ca2+. The SuTC was not affected by limiting the contributions of internal Ca2+ sources. This suggests that the SuTC in rat bladders is unrelated to known Ca2+ mobilization mechanisms.
Oh
S J
SJ
Department of Urology, Seoul National University College of Medicine, Korea.
Ahn
S C
SC
Kim
S J
SJ
Kim
K W
KW
Lee
A
A
Kim
K M
KM
Choi
H
H
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology*; D005260:Female; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects*; D010277:Parasympathomimetics / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D001743:Urinary Bladder / Q000187:drug effects*
D002121:Calcium Channel Blockers; D010277:Parasympathomimetics; D002217:Carbachol; D002118:Calcium