Characterization of the properties of canine colonic smooth muscle in culture.
We have developed and characterized an organ culture system that maintains the viability of colonic smooth muscles. Morphological, mechanical, electrical, and molecular properties of cultured canine colonic circular muscles were determined. Strips of circular muscle were cultured for up to 6 days. The smooth muscle phenotype was retained during culture; muscles contracted to agonists and responded to electrical field stimulation, suggesting that intrinsic nerves also survived in culture. Morphological analysis showed identifiable smooth muscle cells, enteric neurons, and interstitial cells, but some alterations in ultrastructure were also observed. Mechanical responses to acetylcholine suggested that the muscles developed supersensitivity during the culture period. The resting membrane potentials of cells near the submucosal surface of the circular muscle layer decreased from -82 mV on day 0 to -55 mV on day 3. Similar changes in the resting potential gradient occur when colonic muscles are treated with inhibitors of the Na(+)-K(+)-ATPase. Resting potentials of day 3 muscles remained constant in low external K+ (0.1 mM), suggesting little contribution of the pump to resting potential. Northern analysis of RNA from muscles cultured up to 6 days showed that the alpha 2-isoform of the pump decreased. The data suggest that organ-cultured strips of smooth muscle may provide a useful tool for evaluating electrical and mechanical events in conjunction with molecular analysis of functional components.