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1 Department of Pharmacology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA; Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
2 Department of Pharmacology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
* To whom correspondence should be addressed. E-mail: jmbeckel{at}pitt.edu.
Although nicotinic acetylcholine receptors in both the central and peripheral nervous
systems play a prominent role in the control of urinary bladder function, little is known
regarding expression or function of nicotinic receptors in the bladder epithelium, or
urothelium. Nicotinic receptors have been described in epithelial cells lining the upper
GI tract, respiratory tract, and the skin. Thus, the present study examined the expression
and functionality of nicotinic receptors in the urothelium, as well as the effects of
stimulation of nicotinic receptors on the micturition reflex. mRNA for the 
3, 5, 7, 
3,
and 4 nicotinic subunits was identified in rat urothelial cells using RT-PCR. Western
blot also confirmed urothelial expression of the 3 and 7 subunits. Application of
nicotine (50nM) to cultured rat urothelial cells elicited an increase in [Ca+2]i, indicating
that at least some of the subunits form functional channels. These effects were blocked
by the application of the nicotinic antagonist hexamethonium. During in vivo bladder
cystometrograms in urethane anesthetized rats, intravesical administration of nicotine,
choline or the antagonists methyllycaconitine citrate and hexamethonium elicited changes
in voiding parameters. Intravesical nicotine (50nM, 1µM) increased the intercontraction
interval (ICI). Intravesical choline (1-100µM) also affected bladder reflexes similarly,
suggesting that 7 nicotinic receptors mediate this effect. Intravesical administration of
hexamethonium (1-100µM) potentiated the nicotine-induced changes in bladder reflexes.
Methyllycaconitine citrate, a specific 7-receptor antagonist, prevented nicotine-,
choline- and hexamethonium-induced bladder inhibition. These results are the first
indication that stimulation of non-neuronal nicotinic receptors in the bladder can affect
micturition.
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