Comparison of the contractile and metabolic effects of muscarinic stimulation with those of KCl.
Urinary bladder emptying is mediated primarily by a coordinated contraction of the bladder body in response to parasympathetic stimulation and muscarinic receptor activation. In previous studies we presented evidence that the contractile response to bethanechol stimulation could be dissociated from the metabolic response through the use of diltiazem (calcium channel blockade). The conclusion from these studies was that muscarinic stimulation resulted in a significant increase in metabolic activity which was not directly associated with contraction. KCl stimulates contraction in isolated strips by directly depolarizing the membrane rather than from binding to specific membrane receptors. The current study directly compares the metabolic and contractile activity of bethanechol (muscarinic stimulation) with KCl (direct membrane depolarization). Isolated strips of rabbit urinary bladder body were monitored in vitro for changes in intracellular-free calcium, NADH/NAD ratio, and contraction. Intracellular-free calcium was monitored by preincubation of isolated bladder smooth muscle strips with FURA-2 AM and continuously measuring the fluorescence with an MB2 surface spectrofluorometer using excitation wavelengths of 340 and 380 nm, and an emission wavelength of 510 nm. The NADH/NAD ratio was monitored with the MB2 surface spectrophotometer using an excitation wavelength of 366 nm and an emission wavelength of 450 nm. Contraction was monitored using a isometric force transducer connected to a Grass model D polygraph. The results can be summarized as follows. (1) Both bethanechol and KCl stimulate a sharp decrease in the NADH/NAD ratio, a rapid increase in intracellular-free calcium, and a slower increase in contractile force.(ABSTRACT TRUNCATED AT 250 WORDS)
Levin
R M
RM
Division of Urology, University of Pennsylvania School of Medicine, Philadelphia.
Hypolite
J
J
Longhurst
P A
PA
Wein
A J
AJ
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D001626:Bethanechol Compounds / Q000037:antagonists & inhibitors / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D004110:Diltiazem / Q000494:pharmacology; D005453:Fluorescence; D016257:Fura-2; D066298:In Vitro Techniques; D007537:Isometric Contraction / Q000187:drug effects; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects* / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D010277:Parasympathomimetics / Q000494:pharmacology*; D010725:Phosphocreatine / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology*; D011817:Rabbits; D001743:Urinary Bladder / Q000187:drug effects / Q000378:metabolism
D001626:Bethanechol Compounds; D010277:Parasympathomimetics; D010725:Phosphocreatine; D009243:NAD; D001285:Atropine; D000255:Adenosine Triphosphate; D004110:Diltiazem; D011188:Potassium; D002118:Calcium; D016257:Fura-2