Regulation Of Arterial Tone Research Articles

Neurogenic Regulation of Cerebral Blood Flow

The brain has a well-developed vascular network, which allows it to consume up to 15% of the cardiac output with a low mass relative to the whole-body weight. Normally, the metabolic demands of the brain depend considerably on the intensity of functioning of its different departments, which requires constant regulation of the local blood flow level. On the other hand, the state of systemic hemodynamics can have a significant impact on the organ blood flow. Complex and multilevel mechanisms of the regulation of organ cerebral blood flow are aimed at minimizing the possible adverse effects of systemic hemodynamics impairment. The importance of precise and prompt regulation of cerebral blood flow is reinforced by the absence of energy reserves or substrates for its autonomous production in the nervous tissue. The main mechanisms of cerebral blood flow regulation include: myogenic regulation, influence of local humoral influences and vasoactive substances (hormones, metabolites) of systemic blood flow, changes in blood gas composition (increase or decrease in blood oxygen or carbon dioxide tension). In addition, endothelium-dependent mechanisms of regulation are distinguished. Finally, one more level of cerebral arterial tone regulation is represented by the effect of neurotransmitters released from vasomotor fibers terminals of sympathetic and parasympathetic sections of autonomic nervous system, as well as from subcortical neurons and cortical interneurons terminals. In the present review, the principles of neurogenic regulation of cerebral blood flow are considered. The neurogenic regulation of vascular tone is the most complex regulatory circuit. The autonomic innervation of cerebral vessels has significant features that distinguish it from that in most other organs of the great circulatory circle. In addition to the autonomic innervation proper, the vessels of the brain receive sensory innervation, and the small intracerebral arterioles are also innervated -directly by subcortical neurons and cortical interneurons. In this connection, a deeper understanding of the molecular mechanisms of the neurogenic regulation of cerebral blood flow may serve as a basis for the development of new methods of treatment of severe brain diseases based on neuromodulation in the long term.

R-(+)-WIN55212-2 protects pericytes from ischemic damage and restores retinal microcirculatory patency after ischemia/reperfusion injury

Background and purposeCannabinoids are vasoactive substances that act as key regulators of arterial tone in the blood vessels supplying peripheral tissues and the central nervous system. This study aimed to investigate the potential of R-(+)-WIN55212-2 (WIN), a cannabinoid receptor 1 agonist (CB1), as a treatment for retinal ischemia/reperfusion (I/R) injury. Experimental approachMale Wistar rats were subjected to retinal I/R injury by increasing intraocular pressure in the anterior chamber. The rats were randomly divided into four groups: normal control, I/R, vehicle (pre-treated with dimethyl sulfoxide [DMSO] via intraperitoneal injection), and experimental (pre-treated with WIN at a dose of 1 ml/kg via intraperitoneal injection). The rats were sacrificed at different time points of reperfusion (1 hour, 3 hours, 6 hours, and 1 day) after inducing retinal I/R injury, and their retinas were collected for analysis. Oxygen-glucose deprived/reperfusion (OGD/R) was performed by initially perfusing the retinas with oxygenated artificial cerebrospinal fluid (ACSF), then switching to an OGD solution to simulate ischemia, followed by another perfusion with ACSF. Pericyte contraction and the "no-reflow" phenomenon were observed using infrared differential interference contrast (IR-DIC) microscopy and immunohistochemistry. Western blot, enzyme-linked immunosorbent assay (ELISA), and nitric oxide (NO) detection were used to explore the potential mechanism. Key resultsIn both the OGD/R and I/R models, retinal pericytes exhibited persistent contraction even after reperfusion. The ability of WIN to regulate the tone of retinal pericytes and capillaries was specifically blocked by the BKCa inhibitor iberiotoxin (100 nM). WIN demonstrated a protective effect against retinal I/R injury by preserving blood flow in vessels containing pericytes. Pretreatment with WIN alleviated the persistent contraction and apoptosis of retinal pericytes in I/R-induced rats, accompanied by a reduction in intracellular calcium ion (Ca2+) concentration. The expression of CB1 decreased in a time-dependent manner in the I/R group. After I/R injury, endothelium-derived nitric oxide (eNOS) levels were reduced at all time points, which was successfully reversed by WIN therapy except for the 1 day group. Additionally, the downregulation of cyclic guanosine monophosphate (cGMP) and BKCa expression at 3 hours, 6 hours, and 1 day after I/R injury was restored by pretreatment of WIN. Conclusions & implicationsWIN exerted its protective effects on retinal I/R injury by inhibiting the contraction and apoptosis of pericytes through the CB1-eNOS-cGMP-BKCa signaling pathway, thus ameliorated the occlusion of retinal capillaries.

PS-BPB09-4: ARTERIAL SITES AND SEX DIFFERENCES IN ENHANCING VASORELAXATION RESPONSE BY PERIVASCU-LAR ADIPOSE TISSUE IN METABOLIC SYNDROME RATS

Objective: Regulation of arterial tone by perivascular adipose tissue (PVAT) differs with sex. We have demonstrated that PVAT compensates vascular tone when vascular dysfunc-tion occurs in mesenteric arteries of male SHRSP. Z-Leprfa/IzmDmcr (SPZF) rats, but such compensation function disappears in the late stage of metabolic syndrome (MetS). In con-trast, the favorable effects of PVAT remain in female SPZF even after the effects disappear in age-matched males. Design and method: Therefore, in this study, we investigated whether the sex differences in PVAT response are observed in another arterial sites in SPZF and in another MetS model, SHR/NDmcr-cp (CP) rats as well. Renal arteries were isolated from male and female SPZF rats and CP rats at 23 weeks of age. Ring preparations with and without PVAT were made. Vasodilation and mRNA transcript levels in PVAT were examined using organ bath methods and quantitative real-time PCR, respectively. Results, and Conclusions: In renal arteries without PVAT of SPZF and CP rats, acetylcholine-induced relaxations were smaller in female than in male. The presence of PVAT increased the relaxations in CP rats only. In contrast, sodium nitroprusside-induced relaxations, nevertheless presence or ab-sence of PVAT, were smaller in female than in male in SPZF rats, but sex differences were not observed in CP rats. Apelin mRNA levels in female CP rats were the highest among groups. The enhancements of acetylcholine-induced relaxations by PVAT were positively correlated with apelin mRNA levels in PVAT. These results demonstrated that sex difference in enhancing vasorelaxation response by PVAT in renal arteries differs to that in mesenteric arteries in SPZF rats at the same age and to that in another MetS model strain, CP rats, at the same age and arterial sites. This discrepancy observed in female rats between two MetS strains may be associated with the difference in arterial dysfunction mechanism; namely, impairment of only endothelial nitric oxide production in CP rats, and deterioration of re-sponse to nitric oxide in smooth muscle in SPZF rats. Furthermore, apelin levels of PVAT may be involved in the appearance of modulation of PVAT on arterial tone.

Small extracellular vesicles from plasma of women with preeclampsia increase myogenic tone and decrease endothelium-dependent relaxation of mouse mesenteric arteries.

Preeclampsia (PE) is a common syndrome of pregnancy, characterized by new-onset hypertension and proteinuria after gestational week 20, or new onset of hypertension and significant end-organ dysfunction. In the worst cases, it can threaten the survival of both mother and baby. Extracellular vesicles (EVs) are lipid-bilayer nanoparticles released from cells. They are involved in cell-cell communication and transport of diverse cargo molecules. Small extracellular vesicles (sEVs, exosomes) are defined by their size and biogenesis within the endocytic compartment of the cell or reverse budding of the plasma membrane. The function of circulating gestational EVs, released from maternal organs or the placenta, remains to be explored. Here, we focused on sEVs that circulate in the maternal blood in the third trimester of human pregnancy and hypothesized that sEVs from pregnant women with PE play a role in regulation of vessel tone. When compared to sEVs from women with uncomplicated pregnancies, ex vivo exposure of isolated mouse mesenteric arteries to sEVs purified from the plasma of pregnant women with PE led to constriction in response to intraluminal pressure. This effect was not observed using microvesicles from the plasma of women with PE or using PE plasma that was depleted of EVs. Blood vessels exposed to sEVs from women with PE were also more resistant to methacholine-stimulated relaxation. Immunofluorescence microscopy confirmed the presence of sEVs within the vessel wall. Together, these data support the notion that circulating sEVs from pregnant women play a role in the regulation of arterial tone.

Perivascular Adipose Tissue Compensation for Endothelial Dysfunction in the Superior Mesenteric Artery of Female SHRSP.Z-Leprfa/IzmDmcr Rats

Regulation of arterial tone by perivascular adipose tissue (PVAT) differs between sexes. In male SHRSP.Z-Lepr^fa/IzmDmcr rats (SHRSP.ZF), PVAT exerts a compensatory relaxation effect for the loss of endothelium-mediated vasorelaxation, which occurs during the early stages of metabolic syndrome. However, this effect deteriorates by 23 weeks of age. Here, therefore, we compared the effects of PVAT in female and male SHRSP.ZF. Acetylcholine-induced relaxation in superior mesenteric artery without PVAT did not differ between 23-week-old females and males. However, the presence of PVAT enhanced relaxation in 23-week-old females, but not in males. The mRNA levels of angiotensin II type 1 receptor (AT1R) in PVAT did not differ between sexes, but AT1R-associated protein (ATRAP) and apelin levels were higher in females than in males. We observed a positive relationship between differences in artery relaxation with and without PVAT and ATRAP or apelin mRNA levels. In 30-week-old females, PVAT-enhanced relaxation disappeared, and mRNA levels of AT1R increased, while apelin levels decreased compared to 23-week-old females. These results demonstrated that in SHRSP.ZF, PVAT compensation for endothelium dysfunction extended to older ages in females than in males. Apelin and AT1R/ATRAP expression in PVAT may be predictors of favorable effects.

Effect of age on regulation of arterial tone by perivascular adipose tissue in rats with metabolic syndrome

Effect of age on regulation of arterial tone by perivascular adipose tissue in rats with metabolic syndrome

Abstract P171: Comparison Of Modulation Of Renal Arterial Tone By Perivascular Adipose Tissue Between Two Animal Models Of Metabolic Syndrome

Sex differences have recently been noticed in the regulation of arterial tone by perivascular adipose tissue (PVAT). In SHRSP.Z- Lepr fa /IzmDmcr (SHRSP.ZF) rats, an animal model of metabolic syndrome (MetS), we demonstrated that mesenteric and renal PVAT in female rats consistently have an enhancing effect on vasodilation at 23 weeks, an age by which the effect of PVAT is impaired in the male rats. This could explain the sex differences in the prevalence of cardiovascular complications in patients with MetS. Therefore, we determined whether the sex difference in PVAT response also occurs in another animal model of MetS, SHR/NDmcr-cp (SHR-cp) rats.Renal arteries were isolated from male and female 23-week-old SHR-cp rats, and ring preparations with and without PVAT were made. After a stable contraction was obtained by phenylephrine administration, vasodilation in response to acetylcholine was examined using organ bath methods.Vasodilation in arteries without PVAT from female rats was smaller than that in arteries without PVAT from male rats, and presence of PVAT in arteries from female rats increased vasodilation to the same level as that observed in arteries without PVAT from male rats. Furthermore, renal PVAT in male rats was shown to have an enhancing effect on vasodilation.The present study did not identify sex differences in renal PVAT-mediated modulation in SHR-cp rats because the enhancing effects of PVAT did not disappear in male SHR-cp rats, in contrast to that observed in male SHRSP.ZF rats at the same age. The difference in PVAT response in male rats between two MetS models may be due to differences in the severity of MetS symptoms, especially blood pressure, between the models.

Remodeling of Arterial Tone Regulation in Postnatal Development: Focus on Smooth Muscle Cell Potassium Channels.

Maturation of the cardiovascular system is associated with crucial structural and functional remodeling. Thickening of the arterial wall, maturation of the sympathetic innervation, and switching of the mechanisms of arterial contraction from calcium-independent to calcium-dependent occur during postnatal development. All these processes promote an almost doubling of blood pressure from the moment of birth to reaching adulthood. This review focuses on the developmental alterations of potassium channels functioning as key smooth muscle membrane potential determinants and, consequently, vascular tone regulators. We present evidence that the pattern of potassium channel contribution to vascular control changes from Kir2, Kv1, Kv7 and TASK-1 channels to BKCa channels with maturation. The differences in the contribution of potassium channels to vasomotor tone at different stages of postnatal life should be considered in treatment strategies of cardiovascular diseases associated with potassium channel malfunction.

CACNA1C rs1006737 SNP increases the risk of essential hypertension in both Chinese Han and ethnic Russian people of Northeast Asia.

Voltage-gated Ca2+ channels play a key role in the regulation of arterial tone and blood pressure. The aim of this study was to determine whether the association of calcium voltage-gated channel subunit alpha1 C (CACNA1C) rs1006737 with essential hypertension (EH) exists in both Chinese Han and ethnic Russian populations of Northeast Asia. We used a case-control study of 2 ethnic groups in the same latitude geographical area to investigate the association between the susceptibility of EH and rs1006737 polymorphism. A total of 1512 EH patients and 1690 controls in Chinese Han people (Heilongjiang Provence, China), 250 EH patients, and 250 controls in ethnic Russian people (Chita, Russia), participated in this study. All participants were genotyped using the TaqMan SNP genotyping assay (Agena Company). Baseline characteristics and the minor allele frequencies of rs1006737 vary substantially among common Chinese Han and ethnic Russian people. Allele A was found to be a risk factor for EH in Chinese Han [(odds ratio) OR 1.705, (confidence interval) 95% CI: 1.332–2.182, P < .001] and ethnic Russian (OR 1.437; 95% CI: 1.110–1.860, P = .006). The GA genotype was significantly associated with an increased risk of hypertension (OR 1.538, 95% CI: 1.188–1.991, P = .001) for Chinese Han people, and the AA genotype (OR 2.412, 95% CI: 1.348–4.318, P = .003) for ethnic Russian people. The results of this study indicate that the A allele of the variant rs1006737 in the CACNA1C gene may be a useful genetic marker for EH risk prediction in Chinese Han and ethnic Russian populations.

Regulation of Arterial Tone in Rats Fed a Long-Term High-Salt Diet

Regulation of Arterial Tone in Rats Fed a Long-Term High-Salt Diet

A stepwise approach to resolving small ionic currents in vascular tissue.

Arterial membrane potential (Vm) is set by an active interplay among ion channels whose principal function is to set contractility through the gating of voltage-operated Ca2+ channels. To garner an understanding of this electrical parameter, the activity of each channel must be established under near-physiological conditions, a significant challenge given their small magnitude. The inward rectifying K+ (KIR) channel is illustrative of the problem, as its outward "physiological" component is almost undetectable. This study describes a stepwise approach to dissect small ionic currents at physiological Vm using endothelial and smooth muscle cells freshly isolated from rat cerebral arteries. We highlight three critical steps, beginning with the voltage clamping of vascular cells bathed in physiological solutions while maintaining a giga-ohm seal. KIR channels are then inhibited (micromolar Ba2+) so that a difference current can be created, once Ba2+ traces are corrected for the changing seal resistance and subtle instrument drift, pulling the reversal potential rightward. The latter is a new procedure and entails the alignment of whole cell current traces at a voltage where KIR is silent and other channels exhibit limited activity. We subsequently introduced corrected and uncorrected currents into computer models of the arterial wall to show how these subtle adjustments markedly impact the importance of KIR in Vm and arterial tone regulation. We argue that this refined approach can be used on an array of vascular ion channels to build a complete picture of how they dynamically interact to set arterial tone in key organs like the brain.NEW & NOTEWORTHY This work describes a stepwise approach to resolve small ionic currents involved in controlling Vm in resistance arteries. Using this new methodology, we particularly resolved the outward component of the KIR current in native vascular cells, voltage clamped in near-physiological conditions. This novel approach can be applied to any other vascular currents and used to better interpret how vascular ion channels cooperate to control arterial tone.

TMEM16A is implicated in the regulation of coronary flow and is altered in hypertension

Coronary artery disease leads to ischaemic heart disease and ultimately myocardial infarction. Thus, it is important to determine the factors that regulate coronary blood flow. Ca2+ -activated chloride channels contribute to the regulation of arterial tone; however, their role in coronary arteries is unknown. The aim of this study was to investigate the expression and function of the main molecular correlate of Ca2+ -activated chloride channels, TMEM16A, in rat coronary arteries. We performed mRNA and protein analysis, electrophysiological studies of coronary artery myocytes, and functional studies of coronary artery contractility and coronary perfusion, using novel inhibitors of TMEM16A. Furthermore, we assessed whether any changes in expression and function occurred in coronary arteries from spontaneously hypertensive rats (SHRs). TMEM16A was expressed in rat coronary arteries. The TMEM16A-specific inhibitor, MONNA, hyperpolarised the membrane potential in U46619. MONNA, T16Ainh -A01, and Ani9 attenuated 5-HT/U46619-induced contractions. MONNA and T16Ainh -A01 also increased coronary flow in Langendorff perfused rat heart preparations. TMEM16A mRNA was increased in coronary artery smooth muscle cells from SHRs, and U46619 and 5-HT were more potent in arteries from SHRs than in those from normal Wistar rats. MONNA diminished this increased sensitivity to U46619 and 5-HT. In conclusion, TMEM16A is a key regulator of coronary blood flow and is implicated in the altered contractility of coronary arteries from SHRs.

Role of BKCa channels in smooth muscle cells of mesenteric artery from type 1 diabetic rats

High conductance calcium‐activated potassium channels (BKCa) play a very important role in the regulation of arterial vascular tone through the hyperpolarization of the cell membrane. Diverse studies in blood vessels from different vascular beds in animal models with type I and II diabetes mellitus (DM) as well as in patients with type II diabetes have shown a decrease in the BK channels functional response which is probably related with cardiovascular complications secondary to this pathology that is the main cause of mortality in these patients.ObjectiveTo explore the expression and role of the BKCa channels in the contractility of the Mesenteric artery of Type I diabetic rats induced with streptozotocin.MethodsWe conducted isometric tension experiments in mesenteric artery rings (&gt;500) to evaluate the function of the BKCa channel when pharmacologically activated with NS1619, a specific activator drug for this channel, or inhibiting it with Iberotoxin (Ibtx) a toxin that specifically blocks these channels. And with the purpose of knowing if BKCa channels are expressed in these arterial beds we conducted inmunofluorescence studies.ResultsWe observerd a small but not significant response to both activator and inhibitor of the BKCa channels in both control and experimental groups. The inmunofluorescence images indicated that the alfa subunit (the pore subunit) and beta 1 (modulator of the BKCa channel activity) are mostly expressed intracelullarly in endothelial cells and in the smooth muscle of these vessels. This suggests that the lack of response to NS1619 and IBtx is due to the fact that the channel is intracelullar and therefore its participation in the determination of muscle tone of the mesenteric artery is not significant. In contrast, preliminary results of isometric tension in first‐order rat mesenteric artery rings (approximately 400 μm in diameter), show a great participation of the BKCa channels in the regulation of the arterial tone of these vessels, by diminishing in about 80% the tension originated when contracting said rings with phenylephrine.ConclusionsBased on these results, we can conclude at present that the BKCa channels have a greater participation in the maintenance of muscle tone of small arteries (resistance) but not in big arteries (conductive) and that the type I diabetes does not affect the BKCa function in large conduncance arteries.Support or Funding InformationSupported by Conacyt 223350 to RETThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Palmitic Acid Methyl Ester and Its Relation to Control of Tone of Human Visceral Arteries and Rat Aortas by Perivascular Adipose Tissue.

Background: Perivascular adipose tissue (PVAT) exerts anti-contractile effects on visceral arteries by release of various perivascular relaxing factors (PVRFs) and opening voltage-gated K+ (Kv) channels in vascular smooth muscle cells (VSMCs). Palmitic acid methyl ester (PAME) has been proposed as transferable PVRF in rat aorta. Here, we studied PVAT regulation of arterial tone of human mesenteric arteries and clarified the contribution of Kv channels and PAME in the effects.Methods: Wire myography was used to measure vasocontractions of mesenteric artery rings from patients undergoing abdominal surgery. Isolated aortic rings from Sprague-Dawley rats were studied for comparison. PVAT was either left intact or removed from the arterial rings. Vasocontractions were induced by external high K+ (60 mM), serotonin (5-HT) or phenylephrine. PAME (10 nM−3 μM) was used as vasodilator. Kv channels were blocked by XE991, a Kv7 (KCNQ) channel inhibitor, or by 4-aminopyridine, a non-specific Kv channel inhibitor. PAME was measured in bathing solutions incubated with rat peri-aortic or human visceral adipose tissue.Results: We found that PVAT displayed anti-contractile effects in both human mesenteric arteries and rat aortas. The anti-contractile effects were inhibited by XE991 (30 μM). PAME (EC50 ~1.4 μM) was capable to produce relaxations of PVAT-removed rat aortas. These effects were abolished by XE991 (30 μM), but not 4-aminopyridine (2 mM) or NDGA (10 μM), a lipoxygenases inhibitor. The cytochrome P450 epoxygenase inhibitor 17-octadecynoic acid (ODYA 10 μM) and the soluble epoxide hydrolase inhibitor 12-(3-adamantan-1-ylureido)-dodecanoic acid (AUDA 10 μM) slightly decreased PAME relaxations. PAME up to 10 μM failed to induce relaxations of PVAT-removed human mesenteric arteries. 5-HT induced endogenous PAME release from rat peri-aortic adipose tissue, but not from human visceral adipose tissue.Conclusions: Our data also suggest that Kv7 channels are involved in the anti-contractile effects of PVAT on arterial tone in both rat aorta and human mesenteric arteries. PAME could contribute to PVAT relaxations by activating Kv7 channels in rat aorta, but not in human mesenteric arteries.

Antenatal/early postnatal hypothyroidism alters arterial tone regulation in 2-week-old rats.

The mechanisms of vascular alterations resulting from early thyroid hormones deficiency are poorly understood. We tested the hypothesis that antenatal/early postnatal hypothyroidism would alter the activity of endothelial NO pathway and Rho-kinase pathway, which are specific for developing vasculature. Dams were treated with propylthiouracil (PTU, 7 ppm) in drinking water during gestation and 2 weeks after delivery, and their progeny had normal body weight but markedly reduced blood levels of thyroid hormones (ELISA). Small arteries from 2-week-old male pups were studied using wire myography, qPCR and Western blotting. Mesenteric arteries of PTU pups, compared to controls, demonstrated smaller maximum response to α1-adrenergic agonist methoxamine and reduced mRNA contents of smooth muscle differentiation markers α-actin and SERCA2A. Inhibition of basal NO synthesis by l-NNA led to tonic contraction of mesenteric arteries and augmented their contractile responses to methoxamine; both l-NNA effects were impaired in PTU pups. PTU pups demonstrated lower blood level of NO metabolites compared to control group (Griess reaction). Rho-kinase inhibitor Y27632 strongly reduced mesenteric arteries responses to methoxamine in PTU pups, that was accompanied by elevated Rho-kinase content in their arteries in comparison to control ones. Unlike mesenteric, saphenous arteries of PTU pups, compared to controls, had no changes in α-actin and SERCA2A contents and in responses to l-NNA and Y27632. In conclusion, thyroid hormones deficiency suppresses the anticontractile effect of NO and potentiates the procontractile Rho-kinase effects in mesenteric arteries of 2-week-old pups. Such alterations disturb perinatal cardiovascular homeostasis and might lead to cardiovascular pathologies in adulthood.

Cannabinoids Regulate the Diameter of Pericyte-Containing Retinal Capillaries in Rats

Background/Aims: Cannabinoids are vasoactive substances that act as key regulators of arterial tone in the blood vessels supplying peripheral tissues and the central nervous system. We therefore investigated the effect of cannabinoids on retinal capillaries and pericytes. Methods: The effects of cannabinoids on capillary diameters were determined using an ex vivo whole-mount rat retinal model. Western blotting, quantitative PCR, and immunohistochemistry were performed to explore the underlying mechanism. Results: Endogenous cannabinoid 2-arachidonoylglycerol and anandamide and exogenous cannabinoid (R-(+)-WIN55212-2) dilated the noradrenaline-precontracted capillaries in a concentration-dependent manner (1 µM to 0.1 mM). The extent of vasorelaxation was positively correlated with changes in pericyte width. The effects of R-(+)-WIN55212-2 on vasorelaxation and pericyte width were inhibited by a cannabinoid receptor type-1 (CB1) antagonist, AM251 or rimonabant (SR141716A), the nitric oxide synthase inhibitor l-NAME, and the guanylate cyclase inhibitor ODQ. They were also abolished by the removal of the endothelium, but not by the cannabinoid receptor-2 antagonist SR144528, the endothelial cannabinoid receptor antagonist O-1918, or the cyclooxygenase inhibitor indomethacin. Conclusion: The exogenous cannabinoid R-(+)-WIN55212-2 promotes the vasorelaxation of pericyte-containing rat retinal capillaries. This effect of R-(+)-WIN55212-2 is dependent on CB₁ and the nitric oxide–cyclic guanosine monophosphate pathway, and requires an intact endothelium.

Do KV 7.1 channels contribute to control of arterial vascular tone?

KV 7.1 voltage-gated potassium channels are expressed in vascular smooth muscle cells (VSMC) of diverse arteries, including mesenteric arteries. Based on pharmacological evidence using R-L3 (KV 7.1 channel opener), HMR1556, chromanol 293B (KV 7.1 channel blockers), stimulation of these channels has been suggested to evoke profound relaxation in various vascular beds of rats. However, the specificity of these drugs in vivo is uncertain. We used Kcnq1-/- mice and pharmacological tools to determine whether KV 7.1 channels play a role in the regulation of arterial tone. R-L3 produced similar concentration-dependent relaxations (EC50 ~1.4μM) of arteries from wild-type (Kcnq1+/+ ) and Kcnq1-/- mice, pre-contracted with either phenylephrine or 60mM KCl. This relaxation was not affected by 10μM chromanol 293B, 10μM HMR1556 or 30μM XE991 (pan-KV 7 channel blocker). The anti-contractile effects of the perivascular adipose tissue (PVAT) were normal in Kcnq1-/- arteries. Chromanol 293B and HMR1556 did not affect the anti-contractile effects of (PVAT). Isolated VSMCs from Kcnq1-/- mice exhibited normal peak KV currents. The KV 7.2-5 channel opener retigabine caused similar relaxations in Kcnq1-/- and wild-type vessels. We conclude that KV 7.1 channels were apparently not involved in the control of arterial tone by α1 -adrenoceptor agonists and PVAT. In addition, R-L3 is an inappropriate pharmacological tool for studying the function of native vascular KV 7.1 channels in mice.

Adipose-Vascular Coupling and Potential Therapeutics.

Excess visceral adipose tissue is associated with increased risk of high blood pressure, lipid disorders, type 2 diabetes, and cardiovascular disease. Adipose tissue is an endocrine organ with multiple humoral and metabolic roles in regulating whole-body physiology. However, perivascular adipose tissue (PVAT) also plays a functional role in regulating the contractile state of the underlying smooth muscle cell layer. Work during the past decade has shown that this adipose-vascular coupling is achieved by production of numerous substances released from PVAT. Animal disease models have been instrumental in identifying biological and pathophysiological functions of this regulation. These studies have produced strong evidence that alterations in the paracrine control of PVAT in the regulation of arterial tone contribute to vascular dysfunction in obesity, hypertension, and cardiometabolic disease. Perivascular relaxing factors, or perhaps their putative targets, might represent exciting new targets for the prevention and treatment of cardiovascular and metabolic diseases.

The Role of DPO-1 and XE991-Sensitive Potassium Channels in Perivascular Adipose Tissue-Mediated Regulation of Vascular Tone.

The anti-contractile effect of perivascular adipose tissue (PVAT) is an important mechanism in the modulation of vascular tone in peripheral arteries. Recent evidence has implicated the XE991-sensitive voltage-gated KV (KCNQ) channels in the regulation of arterial tone by PVAT. However, until now the in vivo pharmacology of the involved vascular KV channels with regard to XE991 remains undetermined, since XE991 effects may involve Ca2+ activated BKCa channels and/or voltage-dependent KV1.5 channels sensitive to diphenyl phosphine oxide-1 (DPO-1). In this study, we tested whether KV1.5 channels are involved in the control of mesenteric arterial tone and its regulation by PVAT. Our study was also aimed at extending our current knowledge on the in situ vascular pharmacology of DPO-1 and XE991 regarding KV1.5 and BKCa channels, in helping to identify the nature of K+ channels that could contribute to PVAT-mediated relaxation. XE991 at 30 μM reduced the anti-contractile response of PVAT, but had no effects on vasocontraction induced by phenylephrine (PE) in the absence of PVAT. Similar effects were observed for XE991 at 0.3 μM, which is known to almost completely inhibit mesenteric artery VSMC KV currents. 30 μM XE991 did not affect BKCa currents in VSMCs. Kcna5−/− arteries and wild-type arteries incubated with 1 μM DPO-1 showed normal vasocontractions in response to PE in the presence and absence of PVAT. KV current density and inhibition by 30 μM XE991 were normal in mesenteric artery VSMCs isolated from Kcna5−/− mice. We conclude that KV channels are involved in the control of arterial vascular tone by PVAT. These channels are present in VSMCs and very potently inhibited by the KCNQ channel blocker XE991. BKCa channels and/or DPO-1 sensitive KV1.5 channels in VSMCs are not the downstream mediators of the XE991 effects on PVAT-dependent arterial vasorelaxation. Further studies will need to be undertaken to examine the role of other KV channels in the phenomenon.

Pannexin channels do not influence endothelial function in porcine coronary arteries

Pannexins are a newly discovered family of proteins with sequence homology to innexins (Billaud et al., 2011). The pannexin family consists of Panx1, Panx2 and Panx3. These proteins form channels in cell membranes to allow ATP to pass through to act as an intercellular messenger. It has recently been suggested that Panx1 plays an important role in endothelium‐dependent regulation of arterial tone, by facilitating endothelium‐dependent hyperpolarizing (EDH) responses (Gaynullina et al., 2015). This study aimed to determine the influence of Panx1 on endothelium‐dependent vasorelaxation induced by bradykinin (BK) in porcine coronary arteries (PCA).PCAs were dissected from pig hearts obtained from a local abattoir. Segments of PCA were prepared for isometric tension recording in oxygenated Krebs solution warmed to 37°C. After assessing tissue contractility with KCl (60 mM), tone was raised to 50% of the KCl response using U46619. Thereafter, responses to the endothelium‐dependent vasorelaxant, BK, were assessed in the absence and presence of the pannexin inhibitors, carbenoxolone (100 μM), mefloquine (20 μM) or probenecid (1 mM). Similar experiments were carried out after treating the PCA with the nitric oxide synthase inhibitor L‐NAME (100 μM) and cyclooxygenase inhibitor, indomethacin (10 μM). Vasorelaxant responses were expressed as a percentage of U46619‐induced contraction. Data were analysed by two‐way ANOVA followed by a Bonferroni post‐hoc test for comparison of multiple groups. P&lt;0.05 was considered significant.Under control conditions, BK produced a concentration‐dependent vasorelaxation (‐log EC50 7.79 ± 0.49; Rmax = 82 ± 23%, n = 4). The response to BK was increased in the presence of carbenoxolone (‐log EC50 = 8.31 ± 0.11; Rmax = 102 ± 5%, n = 4), unaffected by probenecid (‐log EC50 = 8.34 ± 0.30; Rmax = 69 ± 9%, n = 4) but reduced by mefloquine (‐log EC50 = 7.05 ± 0.30; Rmax = 63 ± 22%, n = 4). A much greater concentration of U46619 was required to raise tone in the presence of mefloquine (raised from approximately nanomolar to micromolar concentrations). BK caused a concentration‐dependent relaxation in preparations exposed to L‐NAME and indomethacin, indicating the presence of an EDH response (‐log EC50 = 7.58 ± 0.27; Rmax = 53 ± 9%, n = 4). This EDH response was not affected by either carbenoxolone (‐log EC50 = 7.12 ± 1.63; Rmax = 41 ± 29%, n = 4) or probenecid (‐log EC50 = 7.46 ± 0.70; Rmax = 64 ± 22%, n = 4) but reduced by mefloquine (‐log EC50 = 6.87 ± 2.10; Rmax = 29 ± 15%, n = 4).The data show that BK induces vasorelaxation in the PCA, in part, by causing an EDH response and nitric oxide response. The use of three different pannexin inhibitors produced conflicting effects. Only mefloquine inhibited the response to BK, while probenecid had no effect and carbenoxolone enhanced BK effects. Thus we cannot conclude that pannexins are involved in mediating endothelium‐dependent responses in the PCA.Support or Funding InformationRoyal Embassy of Saudi ArabiaCultural Bureau in London630 Chiswick High Road London W4 5RY United Kingdom