Effect of Cholecystokinin-Octapeptide and Cerulein on Phasic and Tonic Components in Ovine Duodenum with Special Reference to the 'Minute Rhythm'

Cholecystokinin (CCK) can affect phasic contractions and the minute rhythm (MR) in ovine duodenum but its effect on the tonic component remains unclear. Thus, the aim of this study was to assess whether the hormone exerts significant changes on phasic and tonic components of the duodenal motor activity and on phasic and tonic components of the duodenal MR. Mechanical and electrical activities of the duodenum were recorded in four sheep before and after slow intravenous cholecystokinin octapeptide (CCK-OP, doses 20, 200 and 2000 ng/kg b.w.) and cerulein (doses 1, 10 and 100 ng/kg b.w.) administration in the course of phase 2b of the migrating motor complex. During 5-20-minute periods the area under contraction curve for phasic, tonic and total motor activity was measured for the whole curve and separately for the MR- related activity. It was found that both CCK peptides stimulate phasic and tonic components of the duodenal motor activity as well as both these components of the duodenal MR. The effect of CCK peptides on the tonic component was stronger than on the phasic component. These effects were similar in non-fasted and fasted animals. CCK-OP evoked slightly greater effect than cerulein. The effects of these CCK peptides on phasic and tonic components of the MR were similar. It is concluded that CCK-OP and cerulein stimulate both phasic and tonic components of the duodenal motor activity and phasic and tonic components related to the MR in sheep.

Membrane depolarization-induced contraction of rat caudal arterial smooth muscle involves Rho-associated kinase

Depolarization of the sarcolemma of smooth muscle cells activates voltage-gated Ca2+ channels, influx of Ca2+ and activation of cross-bridge cycling by phosphorylation of myosin catalysed by Ca2+/calmodulin-dependent myosin light-chain kinase (MLCK). Agonist stimulation of smooth muscle contraction often involves other kinases in addition to MLCK. In the present study, we address the hypothesis that membrane depolarization-induced contraction of rat caudal arterial smooth muscle may involve activation of Rho-associated kinase (ROK). Addition of 60mM K+ to de-endothelialized muscle strips in the presence of prazosin and propranolol induced a contraction that peaked rapidly and then declined to a steady level of force corresponding to approx. 30% of the peak contraction. This contractile response was abolished by the Ca2+-channel blocker nicardipine or the removal of extracellular Ca2+. An MLCK inhibitor (ML-9) inhibited both the phasic and tonic components of K+-induced contraction. On the other hand, the ROK inhibitors Y-27632 and HA-1077 abolished the tonic component of K+-induced contraction, and slightly reduced the phasic component. Phosphorylation levels of the 20-kDa light chain of myosin increased rapidly in response to 60mM K+ and subsequently declined to a steady-state level significantly greater than the resting level. Y-27632 abolished the sustained and reduced the phasic elevation of the phosphorylation of the 20-kDa light chain of myosin, without affecting the K+-induced elevation of cytosolic free Ca2+ concentration. These results indicate that ROK activation plays an important role in the sustained phase of K+-induced contraction of rat caudal arterial smooth muscle, but has little involvement in the phasic component of K+-induced contraction. Furthermore, these results are consistent with inhibition of myosin light-chain phosphatase by ROK, which would account for the sustained elevation of myosin phosphorylation and tension in response to membrane depolarization.

Octopamine modulates the responses and presynaptic inhibition of proprioceptive sensory neurones in the locust Schistocerca gregaria

A multineuronal proprioceptor, the femoral chordotonal organ (feCO), monitors the position and movements of the tibia of an insect leg. Superfusing the locust metathoracic feCO with the neuromodulator octopamine, or the octopamine agonist synephrine, affects the position (tonic) component of the organ's response, but not the movement (phasic) component. Both octopamine and synephrine act with the same threshold (10(-6) mol l-1). Individual sensory neurones that respond tonically at flexed tibial angles show increased tonic spike activity following application of octopamine, but those that respond at extended angles do not. Tonic spiking of phaso-tonic flexion-sensitive neurones is enhanced but their phasic spiking is unaffected. Bath application of octopamine to the feCO increases the tonic component of presynaptic inhibition recorded in the sensory terminals, but not the phasic component. This inhibition should at least partially counteract the increased sensory spiking and reduce its effect on postsynaptic targets such as motor neurones. Furthermore, some phasic sensory neurones whose spiking is not affected by octopamine nevertheless show enhanced tonic synaptic inputs. The chordotonal organ is not known to be under direct efferent control, but its output is modified by octopamine acting on its sensory neurones to alter their responsiveness to mechanical stimuli and by presynaptic inhibition acting on their central branches. The effects of this neuromodulator acting peripherally on sensory neurones are therefore further complicated by indirect interactions between the sensory neurones within the central nervous system. Increases of sensory neurone spiking caused by neuromodulators may not necessarily lead to parallel increases in the responses of postsynaptic target neurones.

Modulation by L-type Ca2+ channels and apamin-sensitive K+ channels of muscarinic responses in cat chromaffin cells

In the perfused cat adrenal gland stimulated with the muscarinic agonist methacholine chloride (100 microM for 3 min), two components were detected in the catecholamine secretory response: 1) an early phasic component that peaked at 300 ng/5 s catecholamine release and 2) a tonic component whose peak was transient and declined to a plateau of about 140 ng/5 s. Apamin (0.1 microM) increased the phasic component to 1,200 ng/5 s and the tonic component to approximately 350 ng/5 s. In single fura 2-loaded cat adrenal chromaffin cells, the cytosolic Ca2+ concentration ([Ca2+]i) also followed a biphasic pattern after stimulation with methacholine. Depletion of extracellular Ca2+ reduced the phasic [Ca2+]i peak by > 50% and the phasic secretory peak by approximately 90%; both the tonic components of [Ca2+]i and secretion were abolished. Depletion of intracellular Ca2+ pools decreased the phasic and tonic components of [Ca2+]i and secretion with respect to control values; however, the phasic components diminished more than the tonic components of [Ca2+]i and secretion. Although 3 microM furnidipine (a dihydropyridine L-type Ca2+ channel blocker) inhibited the phasic component of [Ca2+]i and secretion, its effects were more pronounced on the tonic component. omega-Conotoxin GVIA (1 microM, an N-type Ca2+ channel blocker) did not affect the [Ca2+]i or the methacholine secretory responses. The secretion peak seems to depend on both extracellular free Ca2+ (Cao2+) entry through L-type Ca2+ channels as well as on the mobilization of Ca2+ from intracellular stores; the plateau depends only on Cao2+ entry through L-type Ca2+ channels.(ABSTRACT TRUNCATED AT 250 WORDS)