Ca2 -activated K Currents of Pancreatic Duct Cells in Guinea-pig

There are numerous studies on transepithelial transports in duct cells including Cl and/or HCO3. However, studies on transepithelial K transport of normal duct cells in exocrine glands are scarce. In the present study, we examined the characteristics of K currents in single duct cells isolated from guinea pig pancreas, using a whole-cell patch clamp technique. Both Cl and K conductance were found with KCl rich pipette solutions. When the bath solution was changed to low Cl, reversal potentials shifted to the negative side, 75 4 mV, suggesting that this current is dominantly selective to K. We then characterized this outward rectifying K current and examined its Ca2 dependency. The K currents were activated by intracellular Ca2. 100 nM or 500 nM Ca2 in pipette significantly (P< 0.05) increased outward currents (currents were normalized, 76.8 7.9 pA, n=4 or 107.9 35.5 pA, n=6) at 100 mV membrane potential, compared to those with 0 nM Ca2 in pipette (27.8 3.7 pA, n=6). We next examined whether this K current, recorded with 100 nM Ca2 in pipette, was inhibited by various inhibitors, including Ba2, TEA and iberiotoxin. The currents were inhibited by 40.4 % (n=3), 87.0 % (n=5) and 82.5 % (n=9) by 1 mM Ba2, 5 mM TEA and 100 nM iberiotoxin, respectively. Particularly, an almost complete inhibition of the current by 100 nM iberiotoxin further confirmed that this current was activated by intracellular Ca2. The K current may play a role in secretory process, since recycling of K is critical for the initiation and sustaining of Cl or HCO3 secretion in these cells.

Characterization of Intermediate Conductance K+ Channels in Submandibular Gland Acinar Cells

There are some evidences that K+ efflux evoked by muscarinic stimulation is not mainly mediated by large conductance K+ (BK) channels in salivary gland. In this experiment, we therefore characterised non BK channels in rat submandibular gland acinar cells and examined the possibility of agonist effect on this channel using a patch clamp technique. Two types of K+ channels were observed in these cells. BK channels were observed in 3 cells from total 6 cells and its average conductance was 152+/- 7 pS (n=3). The conductance of the another types of K+ channel was estimated as 71+/-7 pS (n=6). On the basis of the conductance of this channel, we defined this channel as intermediate conductance K+ (IK) channels, which were observed from all 6 cells we studied. When we increased Ca2+ concentration of the bath solution in inside-out mode, the IK channel activity was greatly increased, suggesting this channel is Ca2+ sensitive. We next examined the effect of carbachol (CCh) and isoproterenol on the activity of the IK channels. 10(-5) M isoproterenol significantly increased the open probability (Po) from 0.08+/-0.02 to 0.21+/-0.03 (n=4, P<0.05). Application of 10(-5) M CCh also increased Po from 0.048+/-0.03 to 0.55+/-0.33 (n=5, P<0.05) at the maximum channel activity. The degree of BK channel activation induced by the same concentration of CCh was lower than that of IK channels; Po value was 0.011+/-0.003 and 0.027+/-0.005 in control and during CCh stimulation (n=3), respectively. The result suggests that IK channels exist in salivary acinar cells and its channel activity is regulated by muscaricinic and beta- adrenergic agonist. We conclude that IK channels also play a putative role in secretion as well as the BK channels in rat submandibular gland acinar cells.