Effects of tetraethylammonium ions on frequency-dependent vasopressin release from the rat neurohypophysis.

1. Isolated rat neurohypophyses were fixed by their stalks to a platinum wire electrode and superfused with oxygenated Krebs-HEPES solution. Vasopressin release into the medium was determined by radioimmunoassay. Vasopressin secretion was increased by electrical stimulation at different frequencies (3-30 Hz) and different train lengths (75-900 pulses). The effects of tetraethylammonium (TEA) ions and of enhanced calcium were tested. 2. Electrical stimulation at 7.5 or 15 Hz evoked a markedly larger release of vasopressin than stimulation at 3 Hz. During continuous stimulation at 7.5 and 15 Hz the evoked vasopressin release per pulse declined rapidly, but with similar time constants for both frequencies indicating that the fatigue of the release process was strongly time dependent. The kinetic analysis showed also that the initial release per pulse was identical for 7.5 and 15 Hz stimulation. Nevertheless, with increasing duration, stimulation at 7.5 Hz became less efficient (in terms of release per total stimulus) than stimulation at 15 Hz and this was due to the time-dependent fatigue. 3. TEA (10 mM) increased the release of vasopressin evoked by 3 Hz stimulation much more than that evoked by 15 Hz stimulation resulting in an equieffective activation of release by both stimuli. On the other hand, elevation of the extracellular calcium from 1.2 to 3 mM did not alter the different efficiency of stimuli of 3 and 15 Hz. In the presence of TEA the time-dependent fatigue of the release during continuous stimulation was prevented, but an additional, slower component of the fatigue became apparent which was release or impulse dependent. 4. As prolongation of the action potential by TEA facilitates preferentially the hormone release evoked by low (ineffective) frequencies, it is suggested that a frequency-dependent broadening of action potentials which reportedly occurs on neurosecretory neurones may play an important role in the frequency-dependent facilitation of hormone release from the rat neurohypophysis.