Ca2+-activated Cl− current in sheep lymphatic smooth muscle
Freshly dispersed sheep mesenteric lymphatic smooth muscle cells were studied at 37°C using the perforated patch-clamp technique with Cs+- and K+-filled pipettes. Depolarizing steps evoked currents that consisted ofl-type Ca2+ [ I Ca(L)] current and a slowly developing current. The slow current reversed at 1 ± 1.5 mV with symmetrical Cl− concentrations compared with 23.2 ± 1.2 mV ( n = 5) and −34.3 ± 3.5 mV ( n = 4) when external Cl− was substituted with either glutamate (86 mM) or I− (125 mM). Nifedipine (1 μM) blocked and BAY K 8644 enhanced I Ca(L), the slow-developing sustained current, and the tail current. The Cl− channel blocker anthracene-9-carboxylic acid (9-AC) reduced only the slowly developing inward and tail currents. Application of caffeine (10 mM) to voltage-clamped cells evoked currents that reversed close to the Cl− equilibrium potential and were sensitive to 9-AC. Small spontaneous transient depolarizations and larger action potentials were observed in current clamp, and these were blocked by 9-AC. Evoked action potentials were triphasic and had a prominent plateau phase that was selectively blocked by 9-AC. Similarly, fluid output was reduced by 9-AC in doubly cannulated segments of spontaneously pumping sheep lymphatics, suggesting that the Ca2+-activated Cl− current plays an important role in the electrical activity underlying spontaneous activity in this tissue.