title: Loss of caveolin-1 expression is associated with disruption of muscarinic cholinergic activities in the urinary bladder.
abstract: Caveolin-1 (Cav1), a structural protein of caveolae, plays cell- and context-dependent roles in signal transduction pathway regulation. We have generated a knockout mouse homozygous for a null mutation of the Cav1 gene. Cav1 knockout mice exhibited impaired urinary bladder contractions in vivo during cystometry. Contractions of male bladder strips were evoked with electric and pharmacologic stimulation (5-40 Hz, 1-10 microM carbachol, 10 mM alpha,beta-methylene ATP, 100 mM KCl). Acetylcholine (ACh) and norepinephrine (NE) release from bladder strips were measured with a radiochemical method by incubating the strips with 14C-choline and 3H-NE prior to electric stimulation, whereas ATP release was measured using the luciferin-luciferase assay with a luminometer. A 60-75% decline in contractility was observed when Cav1 knockout muscle strips were stimulated with electric current or carbachol, compared to wildtype muscle strips. No difference in contractility was noted when contractions were evoked either by the purinergic agonist alpha,beta-methylene ATP, or by extracellular potassium. To investigate the relative contribution of non-cholinergic activity to bladder contractility, the amplitude of the electric stimulation-evoked contractions was compared in the presence of the muscarinic antagonist atropine (1 microM). While the non-muscarinic (purinergic) response was unaltered, muscarinic cholinergic response was principally disrupted in Cav1 knockout mice. The loss of Cav1 gene expression was also associated with a 70% reduction in ACh release. NE and ATP release was not altered. It is concluded that the loss of caveolin-1 is associated with disruption of M3 muscarinic cholinergic activity in the bladder. Both pre-junctional (acetylcholine neurotransmitter release from neuromuscular junctions) and post-junctional (M3 receptor-mediated signal transduction in bladder smooth muscles) mechanisms are disrupted, resulting in impaired bladder contraction.
authors.affiliation: Neurourology Laboratory, Scott Department of Urology, Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: H Henry
authors.initials: HH
authors.lastname: Lai
authors.forename: Timothy B
authors.initials: TB
authors.lastname: Boone
authors.forename: Guang
authors.initials: G
authors.lastname: Yang
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Smith
authors.forename: Susanna
authors.initials: S
authors.lastname: Kiss
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Thompson
authors.forename: George T
authors.initials: GT
authors.lastname: Somogyi
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D051242:Caveolin 1; D022461:Caveolins / Q000235:genetics* / Q000502:physiology*; D004558:Electric Stimulation; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D018727:Muscarinic Antagonists / Q000494:pharmacology; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism; D018377:Neurotransmitter Agents / Q000378:metabolism; D009638:Norepinephrine / Q000378:metabolism; D043587:Receptor, Muscarinic M3 / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000187:drug effects*; D013268:Stimulation, Chemical; D001743:Urinary Bladder / Q000187:drug effects / Q000378:metabolism*
chemical_list: C495541:Cav1 protein, mouse; D051242:Caveolin 1; D022461:Caveolins; D018727:Muscarinic Antagonists; D018377:Neurotransmitter Agents; D043587:Receptor, Muscarinic M3; D011976:Receptors, Muscarinic; D001285:Atropine; D000255:Adenosine Triphosphate; D000109:Acetylcholine; D009638:Norepinephrine