Disruption of gallbladder smooth muscle function is an early feature in the development of cholesterol gallstone disease.

UNLABELLED: BACKGROUND; Decreased gallbladder smooth muscle (GBSM) contractility is a hallmark of cholesterol gallstone disease, but the interrelationship between lithogenicity, biliary stasis, and inflammation are poorly understood. We studied a mouse model of gallstone disease to evaluate the development of GBSM dysfunction relative to changes in bile composition and the onset of sterile cholecystitis. METHODS: BALB/cJ mice were fed a lithogenic diet for up to 8 weeks, and tension generated by gallbladder muscle strips was measured. Smooth muscle Ca(2+) transients were imaged in intact gallbladder. KEY RESULTS: Lipid composition of bile was altered lithogenically as early as 1 week, with increased hydrophobicity and cholesterol saturation indexes; however, inflammation was not detectable until the fourth week. Agonist-induced contractility was reduced from weeks 2 through 8. GBSM normally exhibits rhythmic synchronized Ca(2+) flashes, and their frequency is increased by carbachol (3 µm). After 1 week, lithogenic diet-fed mice exhibited disrupted Ca(2+) flash activity, manifesting as clustered flashes, asynchronous flashes, or prolonged quiescent periods. These changes could lead to a depletion of intracellular Ca(2+) stores, which are required for agonist-induced contraction, and diminished basal tone of the organ. Responsiveness of Ca(2+) transients to carbachol was reduced in mice on the lithogenic diet, particularly after 4-8 weeks, concomitant with appearance of mucosal inflammatory changes. CONCLUSIONS & INFERENCES: These observations demonstrate that GBSM dysfunction is an early event in the progression of cholesterol gallstone disease and that it precedes mucosal inflammation.

Expression of sodium channel Nav1.6 in cholinergic myenteric neurons of guinea pig proximal colon.

We wished to establish the functional identity of Na(v)1.6-expressing myenteric neurons of the guinea pig proximal colon by determining the extent of colocalization of Na(v)1.6 and selected neurochemical markers. Na(v)1.6-like immunoreactivity (-li) was primarily localized to the hillock and initial segments of myenteric neurons located near junctions with internodal fiber tracts. Immunoreactivity for Na(v)1.6 was co-localized with choline-acetyltransferase-li, representing 96% of Na(v)1.6-immunoreactive neurons; about 5% of these neurons showed co-localization with calretinin-li, but none with substance-P-li. Cholinergic neurons expressing Na(v)1.6 were amongst the smallest (somal area <300 mum(2)) of all cholinergic myenteric neurons observed. Only three of 234 Na(v)1.6-immunoreactive neurons exhibited nNOS-li, and none co-localized with calbindin-li. These data suggest that Na(v)1.6 is expressed in a small uniform population of cholinergic myenteric neurons that lie within the guinea pig proximal colon and that are likely to function as excitatory motor neurons.

Expression of the sodium channel Na(v)1.2 in chemically identified myenteric neurons in the guinea pig.

Our purpose was to identify Na(v)1.2-expressing myenteric neurons of the small and large intestine of the guinea pig by using antibodies directed against Na(v)1.2 and selected neurochemical markers. Na(v)1.2-like immunoreactivity (-li) co-localized with immunoreactivity for choline acetyltransferase in all regions, representing 45%-67% of Na(v)1.2-positive neurons. Na(v)1.2-li co-localized with immunoreactivity for the neural form of nitric oxide synthase more frequently in the colon (20% of neurons exhibiting Na(v)1.2-li) than in the ileum (8%). Co-localization of Na(v)1.2-li with immunoreactivity for a form of neurofilament (NF145) was infrequently observed in the ileum and colon. Enkephalin-immunoreactive cell bodies co-localized with Na(v)1.2-li in all regions. Few myenteric cell bodies immunoreactive for neuropeptide Y were observed in the ileum, but all co-localized with Na(v)1.2-li. This and our previous data suggest that Na(v)1.2 is widely expressed within the guinea pig enteric nervous system, including the three main classes of myenteric neurons (sensory, motor, and interneurons), and is involved in both excitatory and inhibitory pathways. Notable exceptions include the excitatory motor neurons to the longitudinal smooth muscle, the ascending interneurons of the ileum, and the myenteric neurons immunoreactive for NF145, few of which are immunoreactive for Na(v)1.2.