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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 133 (1993), S. 29-41 
    ISSN: 1432-1424
    Keywords: inwardly and outwardly rectifying K+ currents ; BK channels ; TEA ; Ba2+ ; Cs+ ; sheep parotid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary We have used whole-cell patch-clamp techniques to examine the sensitivities of the inwardly and the outwardly rectifying K+ currents in sheep parotid cells to K+ channel blockers. Extracellular tetraethylammonium (ID50 ≈ 200 μmol/liter), quinine (ID50 ≈ 100 μmol/liter), verapamil (ID50 ≈ 30 μmol/liter) and charybdotoxin (ID50 〈 0.1 μmol/liter) reduced the outwardly rectifying current but had no effect on the inwardly rectifying current. Quinidine inhibited the outwardly rectifying current (ID50 ≈ 200 μmol/liter) and, at a concentration of 1 mmol/liter, reduced the inwardly rectifying current by 35%. Extracellular Ba2+ inhibited both the inwardly and outwardly rectifying K+ currents but the inwardly rectifying K+ current was more sensitive to it (ID50 ≈ 1 μmol/liter) than was the outwardly rectifying K+ current (ID50 ≈ 2 mmol/liter). Extracellular Cs+ reduced the inwardly rectifying K+ current (ID50 ≈ 100 μmol/liter) without affecting the outwardly rectifying current; 4-aminopyridine (1 or 10 mmol/liter), lidocaine (0.1 or 1 mmol/liter) and flecainide (0.01 or 0.1 mmol/liter) affected neither current. In excised outsideout patches, the addition to the bath of quinine (100 μmol/liter), quinidine (100 μmol/liter), verapamil (100 μmol/liter) or charybdotoxin (100 nmol/liter) inhibited Ca2+- and voltage-sensitive 250 pS K+ channels (BK channels), but 4-aminopyridine (1 mmol/ liter) and lidocaine (0.1 mmol/liter) did not. The pattern of blocker sensitivities is thus consistent with the hypothesis that BK channels are responsible for the outwardly rectifying whole-cell current seen in resting sheep parotid cells.
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  • 2
    ISSN: 1432-1424
    Keywords: K+ and Cl− currents ; Tetraethylammonium ; Verapamil ; Quinine ; 4-Aminopyridine ; BK channels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract As in other salivary glands, the secretory cells of the sheep parotid have a resting K+ conductance that is dominated by BK channels, which are activated by acetylcholine (ACh) and are blocked by tetraethylammonium (TEA). Nevertheless, perfusion studies indicate that TEA does not inhibit ACh-evoked fluid secretion or K+ efflux from intact sheep parotid glands. In the present study, we have used whole-cell patch clamp techniques to show that ACh activates K+ and Cl− conductances in sheep parotid secretory cells by increasing intracellular free Ca2+, and we have compared the blocker sensitivity of the ACh-evoked whole-cell K+ current to the previously reported blocker sensitivity of the BK channels seen in these cells. The ACh-induced whole-cell K+ current was not blocked by TEA (10 mmol/l) or verapamil (100 μmol/l), both of which block the resting K+ conductance and inhibit BK channels in these cells. Quinine (1 mmol/l) and quinidine (1 mmol/l), although only weak blockers of the resting K+ conductance, inhibited the ACh-evoked current at 0 mV (K+ current), by 68% and 78%, respectively. 4-Aminopyridine (10 mmol/l) partially inhibited the ACh-induced K+ current and caused it to fluctuate. It also caused the resting membrane currents to fluctuate, possibly by altering cytosolic free Ca2+. Ba2+ (100 μmol/l), a blocker of the inwardly rectifying K+ conductance in sheep parotid cells, had no effect on the ACh-induced K+ current. We conclude that the ACh-induced K+ conductance in sheep parotid cells is pharmacologically distinct from both the outwardly rectifying (BK) K+ conductance and the inwardly rectifying K+ conductance seen in unstimulated cells. Given that in vitro perfusion and K+ efflux studies on other salivary glands in which BK channels dominate the resting conductance (e.g., the rat mandibular, rat parotid and mouse mandibular glands) have revealed an insensitivity to TEA, suggesting that BK channels do not carry the ACh-evoked K+ current, we propose that BK channels do not contribute substantially to the K+ current evoked by ACh in the secretory cells of most salivary glands.
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