Pattern of Ca2+ increase determines the type of secretory mechanism activated in dog pancreatic duct epithelial cells

摘要

Intracellular calcium concentration ([Ca²⁺]i) is a key factor controlling secretion from various cell types. We investigated how different patterns of [Ca²⁺]i signals evoke salt secretion via ion transport mechanisms and mucin secretion via exocytosis in dog pancreatic duct epithelial cells (PDEC). Activation of epithelial P2Y2 receptors by UTP generated two patterns of [Ca²⁺]i change: 2–10 μm UTP induced [Ca²⁺]i oscillations, whereas 100 μm UTP induced a sustained [Ca²⁺]i increase, both in the micromolar range. As monitored by carbon-fibre amperometry, the sustained [Ca²⁺]i increase stimulated a larger increase in exocytosis than [Ca²⁺]i oscillations, despite their similar amplitude. In contrast, patch-clamp recordings revealed that [Ca²⁺]i oscillations synchronously activated a K⁺ current as efficiently as the sustained [Ca²⁺]i increase. This K⁺ current was mediated by intermediate-conductance Ca²⁺-activated K⁺ channels (32 pS at −100 mV) which were sensitive to charybdotoxin and resistant to TEA. Activation of these Ca²⁺-dependent K⁺ channels hyperpolarized the plasma membrane from a resting potential of −40 mV to −90 mV, as monitored in perforated whole-cell configuration, in turn enhancing Na⁺-independent, Cl⁻-dependent and DIDS-sensitive HCO3⁻ secretion, as monitored through changes in intracellular pH. PDEC therefore encode concentrations of purinergic agonists as different patterns of [Ca²⁺]i changes, which differentially stimulate K⁺ channels, the Cl⁻–HCO3⁻ exchanger, and exocytosis. Thus, in addition to amplitude, the temporal pattern of [Ca²⁺]i increases is an important mechanism for transducing extracellular stimuli into different physiological effects.