AIM:During early post-natal development, arterial contraction depends less on Ca2+ -signalling pathways but more on changes in Ca2+ -sensitivity compared to adult animals. Whether this difference is related to Rho-kinase, one of the major players affecting Ca2+ -sensitivity, is unknown for intact vessels. Thus, we tested the hypothesis that Rho-kinase critically contributes to the higher Ca2+ -sensitivity of contraction in intact arteries of 1-week-old rats.METHODS:We studied 1-week-old, 4- to 5-week-old and 10- to 12-week-old rats performing isometric myography, Ca2+ -fluorimetry and Western blotting using intact saphenous arteries and arterial pressure measurements under urethane anaesthesia.RESULTS:In 10- to 12-week-old rats, methoxamine (MX) produced vasoconstriction associated with an increase in [Ca2+ ]i and Ca2+ -sensitivity. In contrast, in 1-week-old rats these contractions were accompanied only by an increase in Ca2+ -sensitivity. All MX-induced effects were reduced by the Rho-kinase inhibitor Y-27632; this reduction was complete only in 1-week-old rats. The Rho-kinase specific site Thr855 on MYPT1 was increasingly phosphorylated by MX in vessels of 1-week-old, but not 10- to 12-week-old rats; this effect was also inhibited completely by Y-27632. The Rho-kinase inhibitor fasudil in a dose not affecting the pressor response to MX in 4- to 5-week-old rats reduced it considerably in 1-week-old rats.CONCLUSION:Our results suggest that the higher Ca2+ -sensitivity of arterial contraction in 1-week-old compared to 10- to 12-week-old rats is due to a greater Rho-kinase activity. Constitutively active Rho-kinase contributes to MX-induced contraction in 10- to 12-week-old rats. In 1-week-old rats, additional Rho-kinase activation is involved. This remodelling of the Rho-kinase pathway is associated with its increased contribution to adrenergic arterial pressure responses.

Aerobic exercise training is associated with adaptive changes in skeletal muscles and their vascular bed; such changes in individual muscles may vary depending on their characteristics and recruitment. This study was aimed at comparing the effects of eight-week treadmill training on the locomotor and respiratory muscles in rats. The training course increased the aerobic performance in rats, which was evidenced by an increase in maximum O2 consumption and a decrease in the blood lactate concentration in ramp test. The succinate dehydrogenase activity was increased in the red portion of the gastrocnemius muscle, but not in the diaphragm of trained rats. Arterial segments were isolated from feed arteries and studied by wire myography. The relaxation in response to acetylcholine in gastrocnemius arteries in trained animals was higher as compared with controls (due to higher NO production), while contractile responses to noradrenaline (in the presence of propranolol) were not changed. On the contrary, the endothelial function of diaphragm arteries was not affected by training, but contractile responses to activation of α-adrenoceptors were markedly increased. Thus, aerobic training may increase the blood supply rate to both locomotor and respiratory muscles, but the underlying regulatory mechanisms are different. The results obtained allow us to reveal the physiological mechanisms that determine the physical performance of the body under conditions of compromised functioning of the respiratory system.

Aim: Protein kinases, activated by vasodilator substances, affect vascular function by regulating large conductance Ca2+-activated K+ (K(Ca)1.1) channels. Thus, the aim of the present investigation was to address the hypothesis that quercetin-induced vasorelaxation is caused by a PKG-mediated stimulation of K(Ca)1.1 currents.Methods: Single freshly isolated myocytes and endothelium-denuded rings of the rat tail main artery were employed for electrophysiological and contractility measurements respectively.Results: Quercetin relaxed vessels and increased K(Ca)1.1 currents in a concentration-dependent manner: both effects were antagonized by the specific K(Ca)1.1 channel blocker iberiotoxin. Stimulation of K(Ca)1.1 currents was fully reversible upon drug washout, markedly reduced by Rp-8-Br-PET-cGMPs, a PKG-inhibitor, but not affected by catalase. Quercetin shifted by 34.3mV the voltage dependence of K(Ca)1.1 channel activation towards more negative membrane potentials without affecting its slope. Under conditions of tight functional coupling between sarcoplasmic reticulum Ca2+ release sites and K(Ca)1.1 channels, quercetin decreased both the frequency and the amplitude of K(Ca)1.1 transient currents in a ryanodine-like manner.Conclusion: The natural flavonoid quercetin relaxes the rat tail main artery partly via a PKG-mediated stimulation of smooth muscle K(Ca)1.1 channels.

Characteristics of the small arteries (with a diameter of 200-250 μm) feeding the medial gastrocnemius muscle and diaphragm were studied. Recording of the mechanical activity of ring segments under isometric conditions demonstrated that, similar to other arteries feeding the muscles with a high content of slow fibers, the diaphragm arteries are highly sensitive to adrenoceptor agonists and acetylcholine. The differences in the endothelium-dependent relaxation in response to acetylcholine were retained in the presence of L-NAME and diclofenac. The diaphragm and gastrocnemius arteries similarly responded to serotonin. On the other hand, a high innervation density was characteristic of the diaphragm arteries unlike the arteries of other slow muscles. The density of adrenergic nerve plexus in the diaphragm arteries was considerably higher than in the gastrocnemius arteries. The results suggest that the characteristics of small diaphragm arteries are determined not only by the oxidative capacity of diaphragm muscle fibers, but also by the fact that this is a respiratory muscle.

The age-related dynamics of the activity of signalling pathways coupled to alpha1-adrenergic receptors and their dependence on the sympathetic innervation of arterial smooth muscle have been studied. The effects of the protein kinase C inhibitor (GF109203X, 10(-6) M) and the Rho-kinase inhibitor (Y27632, 10(-5) M) on the isometric contraction of the rat saphenous artery, induced by the alpha1-adrenoceptor agonist methoxamine, were examined. It was shown that the sensitivity to methoxamine of arteries from 2-week-old rats that are partially innervated was reduced as compared to adults, but the effects of both inhibitors were more prominent. The denervation induced by the excision of sympathetic ganglia increased the arterial sensitivity to methoxamine but was not accompanied by changes in sensitivity to the inhibitors. Therefore, the postnatal development of the arterial smooth muscle is characterized by a decrease in the contribution of protein kinase C and Rho-kinase to the regulation of contraction; however, these changes do not correlate with changes in the sensitivity of arteries to methoxamine and development of sympathetic innervation.

Addition of N-acetylcysteine induced relaxation of the coronary and basilar arteries thus indicating some basilar NO-stores in these vessels. The maximum capacity of the NO-stores was similar in the coronary and the basilar arteries. Following adaptation to hypoxia, however, the depot was much greater in the coronary artery wall. This seems to be connected with different degree of participation of the NO-dependent vasodiatation in implementation of the adaptive response to hypoxia in coronary and cerebral vascular systems.