Electrophysiology of mouse parotid acini: effects of electrical field stimulation and ionophoresis of neurotransmitters.

Abstract

1. Intracellular micro-electrode recordings of membrane potential and input resistance were made from surface acini of mouse parotid glands placed in a Perspex tissue bath through which oxygenated physiological saline solutions were circulated. The acinar cells were stimulated by microionophoresis of both acetylcholine (ACh) and adrenaline (Ad) from extracellular micropipettes, and by electrical field stimulation via a pair of platinum electrodes. 2. The acinar cells had a mean resting membrane potential of -64.9 mV +/- 0.6 S.E. The input resistance of the unstimulated cell was 4.63 M omega +/- 0.19 S.E. In a number of cells spontaneous miniature depolarizations were observed, associated with synchronous reductions in input resistance. 3. The responses to ionophoresis of both ACh and Ad and the response to supra-maximal field stimulation were identical. Stimulation always evoked a marked decrease in input resistance associated with an initial potential change, generally followed by a delayed hyperpolarization during which the input resistance returned to normal. 4. Field-stimulation responses could be evoked to single shock (1-2 msec) and to low frequency (1-4 Hz) stimulation. The latency for this response was 245 msec +/- 12 S.E. 5. The field-stimulation response was shown to be susceptible to blockade of nerve conduction in sodium-free or tetrodotoxin- (TTX-) containing media; and to blockade of neurotransmitter release in calcium-free media. 6. The field-stimulation and ACh responses were recorded at different levels of membrane potential within the same cells by applying either hyperpolarizing or depolarizing direct current through the recording electrode. The membrane potential at which the initial potential change undergoes reversal, i.e. changes from a depolarization to a hyperpolarization, is known as the equilibrium or reversal potential, EFS and EACh respectively. The field-stimulation (FS) and ACh responses underwent simultaneous reversal at about -60 mV, i.e. EFS = EACh. Equilibrium potentials were also determined indirectly by analysis of the responses evoked by each stimulus in the manner described by Trautwein & Dudel (1958). Using this technique the equilibrium potentials of the responses to all three stimuli, field stimulation, ACh and Ad, were again about -60mV, i.e. EFS = EACh = EAd. 7. Both the field-stimulation and ACh responses were abolished by atrophine (10(-6) M) while the response to Ad persisted. Atropine also abolished all spontaneous activity. The alpha-adrenergic blocker phentolamine (10(-5) M) abolished the response to Ad but left the field-stimulation response unaffected. 8. Electrical field stimulation of isolated segments of salivary gland evoked release of endogenous neurotransmitter as a consequence of neural excitation. The technique of field stimulation thus makes it possible to investigate the functional innervation of a gland using the in vitro preparation. In the mouse parotid gland the field stimulus response was mediated by ACh released from parasympathetic nerve endings.