G H Herok1, T J Millar, P J Anderton, D K Martin. 1. Co-operative Research Centre for Eye Research and Technology, University of New South Wales, Kensington, Australia.
Abstract
PURPOSE: To characterize the properties of an inwardly rectifying K+ (KIR) current in fresh, enzymatically isolated acinar cells from the rabbit superior lacrimal gland. METHODS: New Zealand White rabbits of both sexes were killed by injecting 45 mg/kg pentobarbital sodium, and the glands were excised. Single acinar cells were isolated enzymatically from these glands. Standard patch-clamp techniques were used to record ion currents. RESULTS: Hyperpolarizing voltages evoked KIR currents that had a conductance of 2.7 +/- 0.16 nS (n = 6) in the range -50 mV to -160 mV. The KIR current was activated with steep voltage dependence on hyperpolarization, and the conductance was approximately dependent on the square root of the external K+ concentration. Increasing the pipette Ca2+ concentration from 10(-9) M to 10(-6) M increased the conductance to 5.3 +/- 0.45 nS (n = 7). Internal substitution of K+ with various cations gave the following permeability sequence: K+ (1.0) > Rb+ (0.83) > Li+ (0.15). The KIR current was inhibited by Ba2+ (100 microns), tetraethylammonium (10 mM), and Cs+ (5 mM) but was insensitive to 4-aminopyridine (5 mM). The single-channel conductance was 43 +/- 2.7 pS (n = 11), and the relationship between between single-channel conductance (gamma) and external K+ concentration ([K]o) is given by: gamma = 7.04[K]o0.37 (pS, r2 = 0.99, P < 0.05). The relationship between [K]o and zero current potential (Erev) is given by: Erev = 35.5 log[K]o - 77.8 (mV; r2 = 0.99, P < 0.05). CONCLUSIONS: The KIR current identified in these lacrimal acini has a similar dependence on [K]o as other inward rectifiers of excitable tissues and exocrine glands. However, this study highlights that there are interspecies variations and similarities between KIR channels that could be related to their individual physiological roles. The authors' investigations suggest that one role of the KIR channel in the rabbit superior lacrimal gland acinar cells is to set and stabilize the resting membrane potential. However, this KIR channel may also be involved in secretion, as has been shown to occur in the sheep parotid gland.
PURPOSE: To characterize the properties of an inwardly rectifying K+ (KIR) current in fresh, enzymatically isolated acinar cells from the rabbit superior lacrimal gland. METHODS: New Zealand White rabbits of both sexes were killed by injecting 45 mg/kg pentobarbital sodium, and the glands were excised. Single acinar cells were isolated enzymatically from these glands. Standard patch-clamp techniques were used to record ion currents. RESULTS: Hyperpolarizing voltages evoked KIR currents that had a conductance of 2.7 +/- 0.16 nS (n = 6) in the range -50 mV to -160 mV. The KIR current was activated with steep voltage dependence on hyperpolarization, and the conductance was approximately dependent on the square root of the external K+ concentration. Increasing the pipette Ca2+ concentration from 10(-9) M to 10(-6) M increased the conductance to 5.3 +/- 0.45 nS (n = 7). Internal substitution of K+ with various cations gave the following permeability sequence: K+ (1.0) > Rb+ (0.83) > Li+ (0.15). The KIR current was inhibited by Ba2+ (100 microns), tetraethylammonium (10 mM), and Cs+ (5 mM) but was insensitive to 4-aminopyridine (5 mM). The single-channel conductance was 43 +/- 2.7 pS (n = 11), and the relationship between between single-channel conductance (gamma) and external K+ concentration ([K]o) is given by: gamma = 7.04[K]o0.37 (pS, r2 = 0.99, P < 0.05). The relationship between [K]o and zero current potential (Erev) is given by: Erev = 35.5 log[K]o - 77.8 (mV; r2 = 0.99, P < 0.05). CONCLUSIONS: The KIR current identified in these lacrimal acini has a similar dependence on [K]o as other inward rectifiers of excitable tissues and exocrine glands. However, this study highlights that there are interspecies variations and similarities between KIR channels that could be related to their individual physiological roles. The authors' investigations suggest that one role of the KIR channel in the rabbit superior lacrimal gland acinar cells is to set and stabilize the resting membrane potential. However, this KIR channel may also be involved in secretion, as has been shown to occur in the sheep parotid gland.