Inhibition of KATP channels in the rat tail artery by neurally released noradrenaline acting on postjunctional alpha2-adrenoceptors.

In rat tail artery, activation of postjunctional alpha(2)-adrenoceptors by noradrenaline (NA) released from sympathetic axons produces a slow depolarization (NAD) of the smooth muscle through a decrease in K(+) conductance. In this study we used intracellular recording to investigate whether the K(+) channel involved is the ATP-sensitive K(+) (K(ATP)) channel. Changes in membrane resistance were monitored by measuring the time constant of decay of excitatory junction potentials. The K(ATP) channel blockers, glibenclamide (10 microm) and PNU 37883A (5 microm), depolarized the smooth muscle and increased membrane resistance. Conversely, the K(ATP) channel openers, pinacidil (0.1 and 0.5 microm) and levcromakalim (0.1 microm), hyperpolarized the smooth muscle and decreased membrane resistance. Activation of K(ATP) channels with calcitonin gene-related peptide (CGRP; 10 nM) also hyperpolarized the smooth muscle and decreased membrane resistance. The NAD was abolished by both glibenclamide and PNU 37883A but was potentiated by CGRP. However, unlike CGRP, the directly acting K(ATP) channel openers, pinacidil and levcromakalim, inhibited the NAD. The effects of other K(+) channel blockers were also determined. A high concentration of Ba(2+)(1 mM), which would be expected to block K(ATP) channels, abolished the NAD, whereas teteraethylammonium (1 mM) and 4-aminopyridine (1 mM) increased its amplitude. Apamin (0.5 microm) and a lower concentration of Ba(2+) (0.1 mM) did not affect the NAD. These findings indicate that activation of alpha(2)-adrenoceptors by neurally released NA depolarizes the membrane of vascular smooth muscle by inhibiting K(ATP) channels open in the resting membrane.

Differential expression of the mechanosensitive potassium channel TREK-1 in epicardial and endocardial myocytes in rat ventricle.

Mechanoelectric feedback (MEF) is the process by which mechanical forces on the myocardium induce electrical responses. It is thought that MEF is important in controlling the beat to beat force of contraction in the ventricle, in response to fluctuations in load, and it may also play a role in controlling the dispersion of repolarization. The transduction mechanism for MEF is via stretch sensitive ion channels in the surface membrane of myocytes. Two types of stretch sensitive channels have been described; a non-selective cation channel, and a potassium selective channel. TREK-1 is a member of the recently cloned tandem pore potassium channels that has been shown to be mechanosensitive and to be expressed in rat heart. Here we report that the gene expression level of TREK-1, quantified using real-time RT-PCR against glyceraldehyde phosphate dehydrogenase (GAPDH) as a comparator gene, was found to be 0.34 +/- 0.14 in endocardial cells compared to 0.02 +/- 0.02 in epicardial cells (P < 0.05). To confirm that this is reflected in a different current density, whole cell TREK-1 currents, activated by chloroform, were recorded with patch clamp techniques in epicardial and endocardial cells. TREK-1 current density in epicardial and endocardial cells was 0.21 +/- 0.06 pA/pF and 0.8 +/- 0.27 pA/pF, respectively (P

Selective modulation of noradrenaline release by alpha 2-adrenoceptor blockade in the rat-tail artery in vitro.

The effects of blocking alpha(2)-adrenoceptors on noradrenaline (NA) and adenosine 5'-triphosphate (ATP) release from postganglionic sympathetic nerves have been investigated in rat-tail artery in vitro. Continuous amperometry was used to measure NA release and intracellularly recorded excitatory junction potentials (e.j.p.'s) were used to measure ATP release. Application of the alpha(2)-adrenoceptor antagonist, idazoxan (1 microm), increased the amplitude of NA-induced oxidation currents evoked by trains of 10 stimuli at 1 and 10 Hz. In cells deep in the media, idazoxan (1 microm) had no effect on the amplitude of e.j.p.'s evoked by trains of 10 stimuli at 1 and 10 Hz. In cells close to the adventitial - medial border, idazoxan produced a small increase in the amplitude of e.j.p.'s evoked at the end of trains of 10 stimuli at 1 Hz. In tissues pretreated with the neuronal NA uptake inhibitor, desmethylimpramine (0.3 microm), idazoxan (1 microm) markedly increased the amplitude of e.j.p.'s in cells deep in the media. The alpha(2)-adrenoceptor agonist, clonidine (0.5 microm), produced similar reductions in the amplitudes of both NA-induced oxidation currents and e.j.p.'s evoked by 10 stimuli at 1 Hz. These effects of clonidine were reversed by the subsequent addition of idazoxan (1 microm). The release of both NA and ATP is inhibited to a similar extent by activation of prejunctional alpha(2)-adrenoceptors by clonidine. In contrast, endogenously released NA more markedly inhibits NA release. These findings provide further support for the differential modulation of NA and ATP release.