VR1 Receptors Research Articles

Effects of cannabinoid and vanilloid receptor antagonists on nicotine induced relaxation response enhancement in rabbit corpus cavernosum.

Objective(s):Endocannabinoids and nicotine regulate the neurotransmitter release in different central and peripheral synapses. Various studies in the literature demonstrate the interaction between endocannabinoid and nicotinic systems, especially in the central nervous system. The interaction between nicotinic and endocannabinoid systems was investigated in this study. We aimed to show the effects of cannabinoid and vanilloid receptor antagonists on nicotine-induced relaxation response increases in rabbit corpus cavernosum.Materials and Methods:From a total of seven male albino rabbits, three or four equal strips were cut from each corpus cavernosum and inserted in isolated organ baths. Tissues were contracted with phenylephrine (3×10−5 M). After contraction reached a plateau, strips were stimulated with EFS, and with the stabilization of EFS relaxation responses, 10-4 M of nicotine was administered to tissues. After that, in order to investigate the effects of AM251 (CB1 antagonist), AM630 (CB2 inverse agonist) or capsazepine (a vanilloid receptor antagonist) were given to different tissues, after the resting period.Results:Nicotine (10−4 M) increased the EFS-induced relaxation responses (14.60%±2.94%, P<0.05). AM630 decreased the enhancement of nicotine-induced EFS relaxation responses (nicotine 10-4 M enhancement: 17.16%±3.19%; nicotine 10-4 M enhancement in the presence of AM630 10-6 M: 4.44%±3.43% P<0.05; n=6), whereas effects of AM251 and capsazepine were not significant. Conclusion:In the present study, nicotine increased the amplitudes of EFS-induced relaxation responses probably via nicotinic acetylcholine receptors located on the nitrergic nerves of the corpus cavernosum. We showed the role of cannabinoid-like endo-ligands in nicotine-induced enhancement via CB2 receptors but not CB1 and VR1 receptors.

Molecular control limiting sensitivity of sweet taste neurons in Drosophila

To assess the biological value of environmental stimuli, animals' sensory systems must accurately decode both the identities and the intensities of these stimuli. While much is known about the mechanism by which sensory neurons detect the identities of stimuli, less is known about the mechanism that controls how sensory neurons respond appropriately to different intensities of stimuli. The ionotropic receptor IR76b has been shown to be expressed in different Drosophila chemosensory neurons for sensing a variety of chemicals. Here, we show that IR76b plays an unexpected role in lowering the sensitivity of Drosophila sweet taste neurons. First, IR76b mutants exhibited clear behavioral responses to sucrose and acetic acid (AA) at concentrations that were too low to trigger observable behavioral responses from WT animals. Second, IR76b is expressed in many sweet neurons on the labellum, and these neurons responded to both sucrose and AA. Removing IR76b from the sweet neurons increased their neuronal responses as well as animals' behavioral responses to sucrose and AA. Conversely, overexpressing IR76b in the sweet neurons decreased their neuronal as well as animals' behavioral responses to sucrose and AA. Last, IR76b's response-lowering ability has specificity: IR76b mutants and WT showed comparable responses to capsaicin when the mammalian capsaicin receptor VR1 was ectopically expressed in their sweet neurons. Our findings suggest that sensitivity of Drosophila sweet neurons to their endogenous ligands is actively limited by IR76b and uncover a potential molecular target by which contexts can modulate sensitivity of sweet neurons.

Endogenous prostaglandins and afferent sensory nerves in gastroprotective effect of hydrogen sulfide against stress-induced gastric lesions.

Hydrogen sulfide (H2S) plays an important role in human physiology, exerting vasodilatory, neuromodulatory and anti-inflammatory effects. H2S has been implicated in the mechanism of gastrointestinal integrity but whether this gaseous mediator can affect hemorrhagic lesions induced by stress has been little elucidated. We studied the effect of the H2S precursor L-cysteine, H2S-donor NaHS, the H2S synthesizing enzyme (CSE) activity inhibitor- D,L-propargylglycine (PAG) and the gastric H2S production by CSE/CBS/3-MST activity in water immersion and restraint stress (WRS) ulcerogenesis and the accompanying changes in gastric blood flow (GBF). The role of endogenous prostaglandins (PGs) and sensory afferent nerves releasing calcitonin gene-related peptide (CGRP) in the mechanism of gastroprotection induced by H2S was examined in capsaicin-denervated rats and those pretreated with capsazepine to inhibit activity of vanilloid receptors (VR-1). Rats were pretreated with vehicle, NaHS, the donor of H2S and or L-cysteine, the H2S precursor, with or without the concurrent treatment with 1) nonselective (indomethacin) and selective cyclooxygenase (COX)-1 (SC-560) or COX-2 (rofecoxib) inhibitors. The expression of mRNA and protein for COX-1 and COX-2 were analyzed in gastric mucosa pretreated with NaHS with or without PAG. Both NaHS and L-cysteine dose-dependently attenuated severity of WRS-induced gastric lesions and significantly increased GBF. These effects were significantly reduced by pretreatment with PAG and capsaicin denervation. NaHS increased gastric H2S production via CSE/CBS but not 3-MST activity. Inhibition of COX-1 and COX-2 activity significantly diminished NaHS- and L-cysteine-induced protection and hyperemia. NaHS increased expression of COX-1, COX-2 mRNAs and proteins and raised CGRP mRNA expression. These effects of NaHS on COX-1 and COX-2 protein contents were reversed by PAG and capsaicin denervation. We conclude that H2S exerts gastroprotection against WRS-induced gastric lesions by the mechanism involving enhancement in gastric microcirculation mediated by endogenous PGs, sensory afferent nerves releasing CGRP and the activation of VR-1 receptors.

Capsaicin and N-arachidonoyl-dopamine (NADA) decrease tension by activating both cannabinoid and vanilloid receptors in fast skeletal muscle fibers of the frog.

Previous studies have indicated that vanilloid receptor (VR1) mRNA is expressed in muscle fibers. In this study, we evaluated the functional effects of VR1 activation. We measured caffeine-induced contractions in bundles of the extensor digitorum longus muscle of Rana pipiens. Isometric tension measurements showed that two VR1 agonists, capsaicin (CAP) and N-arachidonoyl-dopamine (NADA), reduced muscle peak tension to 57 ± 4 % and 71 ± 3% of control, respectively. The effect of CAP was partially blocked by a VR1 blocker, capsazepine (CPZ), but the effect of NADA was not changed by CPZ. Because NADA is able to act on cannabinoid receptors, which are also present in muscle fibers, we tested the cannabinoid antagonist AM281. We found that AM281 antagonized both CAP and NADA effects. AM281 alone reduced peak tension to 80 ± 6 % of control. With both antagonists, the CAP effect was completely blocked, and the NADA effect was partially blocked. These results provide pharmacological evidence of the functional presence of the VR1 receptor in fast skeletal muscle fibers of the frog and suggest that capsaicin and NADA reduce tension by activating both cannabinoid and vanilloid receptors.

New satiety hormone nesfatin-1 protects gastric mucosa against stress-induced injury: Mechanistic roles of prostaglandins, nitric oxide, sensory nerves and vanilloid receptors

Nesfatin-1 belongs to a family of anorexigenic peptides, which are responsible for satiety and are identified in the neurons and endocrine cells within the gut. These peptides have been implicated in the control of food intake; however, very little is known concerning its contribution to gastric secretion and gastric mucosal integrity. In this study the effects of nesfatin-1 on gastric secretion and gastric lesions induced in rats by 3.5h of water immersion and restraint stress (WRS) were determined. Exogenous nesfatin-1 (5–40μg/kg i.p.) significantly decreased gastric acid secretion and attenuated gastric lesions induced by WRS, and this was accompanied by a significant rise in plasma NUCB2/nefatin-1 levels, the gastric mucosal blood flow (GBF), luminal NO concentration, generation of PGE2 in the gastric mucosa, an overexpression of mRNA for NUBC2 and cNOS, as well as a suppression of iNOS and proinflammatory cytokine IL-1β and TNF-α mRNAs. Nesfatin-1-induced protection was attenuated by suppression of COX-1 and COX-2 activity, the inhibition of NOS with L-NNA, the deactivation of afferent nerves with neurotoxic doses of capsaicin, and the pretreatment with capsazepine to inhibit vanilloid VR1 receptors. This study shows for the first time that nesfatin-1 exerts a potent protective action in the stomach of rats exposed to WRS and these effects depend upon decrease in gastric secretion, hyperemia mediated by COX-PG and NOS-NO systems, the activation of vagal and sensory nerves and vanilloid receptors.

Chronic Opioid Potentiates Presynaptic but Impairs Postsynaptic N-Methyl-d-aspartic Acid Receptor Activity in Spinal Cords: IMPLICATIONS FOR OPIOID HYPERALGESIA AND TOLERANCE

Chronic Opioid Potentiates Presynaptic but Impairs Postsynaptic N-Methyl-d-aspartic Acid Receptor Activity in Spinal Cords: IMPLICATIONS FOR OPIOID HYPERALGESIA AND TOLERANCE

Alteration in Skeletal Muscle Afferents in Rats with Chronic Heart Failure

An exaggerated exercise pressor reflex (EPR) contributes to exercise intolerance and excessive sympatho-excitation in the chronic heart failure (CHF) state. However, the components of this reflex that are responsible for the exaggerated EPR in CHF remain unknown. To determine whether muscle afferent function is altered in CHF, we recorded the discharge of group III and IV afferents in response to static contraction, passive stretch and hindlimb intra-arterial injection of capsaicin in sham and CHF rats. We also investigated the roles of purinergic 2X receptor (P2X) and the transient receptor potential vanilloid 1 (VR1) in mediating the altered sensitivity of muscle afferents. Compared with sham rats, CHF rats exhibited greater responses of group III afferents to contraction and stretch whereas the responses of group IV afferents to contraction and capsaicin were blunted. Hindlimb intra-arterial infusion of pyridoxal phosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS), a P2X antagonist, attenuated the responses of group III afferents to contraction and stretch in CHF rats to a greater extent than in sham rats. Western blot data showed that P2X3 receptors were significantly upregulated in doral root ganglion (DRG) of CHF rats whereas VR1 receptors were significantly downregulated. Immunohistochemical evidence showed that immunostaining of the P2X3 receptors was more intense in both IB4-positive (C-fibre) and NF200-positive (A-fibre) neurons in DRG of CHF rats whereas the immunostaining of the VR1 receptors was decreased in IB4-positive neurons. These data suggest that group III afferents are sensitized whereas group IV afferents are desensitized in CHF, which is related to the dysfunction of P2X and VR1 receptors.

Alteration in skeletal muscle afferents in rats with chronic heart failure

An exaggerated exercise pressor reflex (EPR) contributes to exercise intolerance and excessive sympatho-excitation in the chronic heart failure (CHF) state. However, the components of this reflex that are responsible for the exaggerated EPR in CHF remain unknown. To determine whether muscle afferent function is altered in CHF, we recorded the discharge of group III and IV afferents in response to static contraction, passive stretch and hindlimb intra-arterial injection of capsaicin in sham and CHF rats. We also investigated the roles of purinergic 2X receptor (P2X) and the transient receptor potential vanilloid 1 (VR1) in mediating the altered sensitivity of muscle afferents. Compared with sham rats, CHF rats exhibited greater responses of group III afferents to contraction and stretch whereas the responses of group IV afferents to contraction and capsaicin were blunted. Hindlimb intra-arterial infusion of pyridoxal phosphate-6-azophenyl-2,4-disulfonic acid (PPADS), a P2X antagonist, attenuated the responses of group III afferents to contraction and stretch in CHF rats to a greater extent than in sham rats. Western blot data showed that P2X3 receptors were significantly upregulated in doral root ganglion (DRG) of CHF rats whereas VR1 receptors were significantly downregulated. Immunohistochemical evidence showed that immunostaining of the P2X3 receptors was more intense in both IB4-positive (C-fibre) and NF200-positive (A-fibre) neurons in DRG of CHF rats whereas the immunostaining of the VR1 receptors was decreased in IB4-positive neurons. These data suggest that group III afferents are sensitized whereas group IV afferents are desensitized in CHF, which is related to the dysfunction of P2X and VR1 receptors.

In reference to capsaicin receptors are colocalized with sweet/bitter receptors in the taste sensing cells of circumvallate papillae

Dear Editors,We read with interest the paper ‘‘Capsaicin receptors arecolocalized with sweet/bitter receptors in the taste sensingcells of circumvallate papillae’’ published by Moon et al.[1]. The authors report a double reaction of T1R2-, T1R3-or T2R6 positive vallate cells with anti-VR1 antibodies ofrat circumvallate papillae. They also found that about 60%of VR1 positive cells in the circumvallatae papillae reac-tive with a-gustducin. The authors concluded that capsaicinmay interact with transducin pathways of sweet and bittersensations.Many researchers described the suppressive effects ofcapsaicin on responses to taste stimuli of sweet, bitter, andumami in humans [2–4]. At lower concentrations, capsai-cin can also elicit bitter [4, 5], and sweet sensations [5].Green and Schullery described bitter sensations whencapsaicin was applied to fungiform, foliate, and circum-vallate papillae of the human tongue [4].In our study, capsaicin was applied to the dorsal anteriortongue in healthy subjects. Beside bitter, sour and saltysensations, capsaicin elicited also sweet sensations [5].The presented study reported evidence of VR1 receptorsin taste cells in circumvallate papillae in rats. Based on theauthor’s findings, one might speculate that VR1 receptorscan also be found in taste cells of the fungiform papillae ofthe dorsal anterior tongue even in humans. This mayexplain why some healthy subjects report sweet sensationsafter stimulation with capsaicin at low concentrations.Knowledge of the VR1 receptors of taste cells of fungiformpapillae may also help to explain symptoms after chordatympani nerve (CTN) transection. In patients with tran-sected CTN the capsaicin thresholds were lower at theipsilateral tongue side compared to the contralateral side[6].We would like to congratulate the authors for theirexcellent study because they clearly emphasize the need forfurther research in taste-trigeminal interactions.References

Antinociceptive effect of geranylgeraniol and 6α,7β-dihydroxyvouacapan-17β-oate methyl ester isolated from Pterodon pubescens Benth

BackgroundPterodon pubescens Benth seeds are commercially available in the Brazilian medicinal plant street market. The crude alcoholic extracts of this plant are used in folk medicine as anti-inflammatory, analgesic, and anti-rheumatic preparations. The aim of this study was to evaluate the contribution of geranylgeraniol (C1) and 6α, 7β-dihydroxyvouacapan-17β-oate methyl ester (C2) isolated from Pterodon pubescens Benth. to the antinociceptive activity of the crude extract.ResultsCompounds C1 and C2 demonstrated activity against writhing with intraperitoneal (i.p.) and oral (p.o.) routes, capsaicin (i.p. and p.o.), glutamate (i.p.), and in the hot-plate (p.o.) tests, demonstrating their contribution to the antinociceptive activity of crude Pterodon pubescens Benth extracts. The observed activity of compounds C1 and C2 may be related to vanilloid receptors VR1, and/or glutamate peripheral receptors. In hot-plate model, the antinociceptive activity was maintained when naloxone chloride (opioid antagonist) was administered prior to treatment with compounds suggesting that C1 and C2 (p.o.) do not exert their antinociceptive effects in the hot-plate test via opioid receptors. The findings presented herein also suggest that compounds within the crude Pterodon pubescens Benth. extract may exert a synergistic interactive effect, since the crude extract (300 mg.kg-1) containing lower concentrations of compounds C1 (11.5%- 34.6 mg. kg-1) and C2 (1.5% - 4.7 mg.kg-1) gave statistically the same effect to the pure compounds when tested separately (C1 = C2 = 300 mg.kg-1) in writhing experimental model with p.o. administration. Further studies will be undertaken to establish more specifically the mechanisms of action for compounds C1 and C2. Possible synergistic interactions will be evaluated employing the Isobole method.ConclusionThese results allowed us to establish a relationship between the popular use of Pterodon pubescens seeds for pain relief and the activity of two major compounds isolated from this species which demonstrated antinociceptive activity. Various "in vivo" experimental models corroborate the folk use of this species for different pain and inflammation disorders.

Design principles of sensory receptors

EDITORIAL article Front. Cell. Neurosci., 07 July 2010Sec. Cellular Neurophysiology Volume 4 - 2010 | https://doi.org/10.3389/fncel.2010.00025

Capsaicin receptors are colocalized with sweet/bitter receptors in the taste sensing cells of circumvallate papillae

We examined co-localization of vanilloid receptor (VR1) with sweet receptors T1R2, T1R3, or bitter receptor T2R6 in taste receptor cells of rat circumvallate papillae. Tissue sections of rat circumvallate papillae were doubly reacted with anti-VR1 antibodies and anti-T1R2, anti-T1R3 or anti-T2R6 antibodies, using double-immunofluorescence histochemistry technique. Localizations of VR1, T1Rs and T2R6 in the vallate taste cells containing α-gustducin were also examined. VR1 immunoreactivities (-ir) were observed in subsets of taste cells in the circumvallate papillae, and 96-99% of the vallate taste cells exhibiting T1R2-, T1R3- or T2R6-ir co-exhibited VR1-ir. Approximately half of T2R6-ir cells (~49%), and 50-58% of T1Rs-ir cells, co-exhibited α-gustducin-ir in the vallate taste buds. About 58% of VR1-ir cells in the vallate exhibited α-gustducin-ir as well. Results support the idea that capsaicin may interact with the transduction pathways of sweet and bitter taste stimuli, possibly in mediation of its receptor VR1 localized in taste receptor cells. Additionally, the partial co-localization of α-gustducin with VR1 suggests that a tentative modulatory function of capsaicin in sweet and bitter transductions in the rat circumvallate comprises of both α-gustducin-mediated and non-mediated transduction pathways.

Intra-oral pre-treatment with capsaicin increases consumption of sweet solutions in rats

Sprague-Dawley rats received preference tests for sucrose or saccharin daily following oral treatment with 0.02% capsaicin. Consumed sweet solutions and preference scores increased in capsaicin-treated rats, compared to control rats on the second to fifth exposure period for sucrose and all exposure periods for saccharin. Chow intake was not affected by repeated treatment with capsaicin. Real-time RT-PCR analysis revealed decreased expression of sweet receptors T1R2 and T1R3 as well as capsaicin receptor VR1 in the circumvallate after this repeated oral exposure to capsaicin. VR1 immunoreactivities were also localized in the vallate taste cells by fluorescence immunohistochemistry. Results suggest that decreased expression of sweet receptors in the circumvallate may be related to increased sweet consumption in capsaicin-treated rats; any causal relationship should be further studied. Also, these data suggest that capsaicin may interact with a sweet transduction pathway in the mediation of its receptor VR1 that are located in the vallate taste cells.

P6.4 Autonomic neurons expressing cannabinoid and vanilloid receptors in postnatal development

Cannabinoids are cell membrane-derived signalling molecules that are released from nerves, blood cells and endothelial cells, and have diverse biological effects. Endogenous cannabinoids and their cannabinoid CB1 and CB2 receptors, are present from the early stages of gestation and play a number of vital roles for the developing organism. The majority of afferent neurons have vanilloid VR1 receptors. The aim of this work was to immunocytochemically investigate the development of the neurons containing CB1, CB2 and VR1 receptors in the nodose ganglion of vagus nerve (GNVN), in the spinal ganglia (SG) and sympathetic ganglia of rats of different ages.

Role of endocannabinoids in the regulation of the cortical blood flow

Event Abstract Back to Event Role of endocannabinoids in the regulation of the cortical blood flow Eva Ruisanchez1*, Ishiguro M. Leszl1, Rita Benko1, Zsombor Lacza1, Zoltan Jarai2, Peter Sandor1 and Zoltan Benyo1 1 Institute of Human Physiology and Clinical Experimental Research, Semmelweis University, Hungary 2 1st Department of Internal Medicine, Semmelweis University, Hungary The influence of endocannabinoids on the resting blood flow and on hypoxia/hypercapnia (H/H)-induced vasodilation was investigated in the cerebral cortex with laser-Doppler (LD) flowmetry in anesthetized adult male Wistar rats. Inhibition of endocannabinoid reuptake by AM-404 (10 mg/kg) resulted in a transient increase of the resting cerebrocortical blood flow (CBF), which had a peak (+65±16 %) at 5.2±0.6 minutes and disappeared within 10 minutes. The CBF changes were accompanied by reduction of the ventilation rate as well as by increased arterial CO2 tension (PaCO2) and decreased O2 tension (PaO2). Application of the CB1 receptor antagonist AM-251 (10 mg/kg iv) did not alter baseline CBF but significantly increased the H/H-induced hyperemia: by 30 % during mild H/H (PaO2=75-85, PaCO2=50-60 mmHg), by 22 % during moderate H/H (PaO2=55-65, PaCO2=80-90 mmHg), and by 14 % during severe H/H (PaO2=45-50, PaCO2=90-100 mmHg). In contrast, vehicle of AM-251 had no significant effect. H/H did not change mean arterial pressure in untreated or vehicle-treated animals, although following AM-251 treatment it induced slight hypertension (+11 ± 6 mmHg). Furthermore, AM-251 was able to prevent the effects of AM-404 while the VR1 receptor antagonist capsazepine (20 mg/kg) had no effect. In conclusion our results indicate that endocannabinoids may increase the resting CBF either by a CB1-mediated direct vasodilator mechanism or indirectly via respiratory depression and the consequent changes of the blood gas tensions. Interestingly, activation of CB1 receptors attenuates the H/H induced cerebrocortical hyperaemia by a yet undefined mechanism. Supported by OTKA K62375 and NKTH H07-BEL74286 grants. Conference: 12th Meeting of the Hungarian Neuroscience Society, Budapest, Hungary, 22 Jan - 24 Jan, 2009. Presentation Type: Poster Presentation Topic: Pathophysiology and neurology - degenerative disorders Citation: Ruisanchez E, Leszl IM, Benko R, Lacza Z, Jarai Z, Sandor P and Benyo Z (2009). Role of endocannabinoids in the regulation of the cortical blood flow. Front. Syst. Neurosci. Conference Abstract: 12th Meeting of the Hungarian Neuroscience Society. doi: 10.3389/conf.neuro.01.2009.04.172 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 05 Mar 2009; Published Online: 05 Mar 2009. * Correspondence: Eva Ruisanchez, Institute of Human Physiology and Clinical Experimental Research, Semmelweis University, Budapest, Hungary, ruievi@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Eva Ruisanchez Ishiguro M Leszl Rita Benko Zsombor Lacza Zoltan Jarai Peter Sandor Zoltan Benyo Google Eva Ruisanchez Ishiguro M Leszl Rita Benko Zsombor Lacza Zoltan Jarai Peter Sandor Zoltan Benyo Google Scholar Eva Ruisanchez Ishiguro M Leszl Rita Benko Zsombor Lacza Zoltan Jarai Peter Sandor Zoltan Benyo PubMed Eva Ruisanchez Ishiguro M Leszl Rita Benko Zsombor Lacza Zoltan Jarai Peter Sandor Zoltan Benyo Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

CB1 Cannabinoid Receptor Inhibition: Promising Approach for Heart Failure?

Endocannabinoids are part of a novel signaling system, along with their cellular receptors and several proteins implicated in their synthesis, release, transport, and degradation. The endocannabinoids are endogenous lipid mediators generated by almost all cell types both in the brain and in the peripheral tissues, which have a wide range of biological effects similar to those of marijuana. 2-Arachidonoylglycerol and arachidonoyl ethanolamide (anandamide) are the 2 best characterized endocannabinoids. Their effects are mediated by 2 G protein–coupled receptors, CB1 and CB2, as well as additional, as-yet unidentified ones.1 2-Arachidonoylglycerol binds to both CB2 and CB1 receptors, whereas arachidonoyl ethanolamide has higher affinity for CB1 receptors and may also bind to vanilloid VR1 receptors. The tissue levels of endocannabinoids are determined by the balance between their biosynthesis (involving phospholipase D– and diacyl-glycerol lipase–dependent and other pathways), cellular uptake, and degradation by fatty acid amide hydrolase and/or monoacylglycerol lipases.1,2 Both CB2 and CB1 receptors are negatively coupled to adenylate cyclase through Gi/o proteins but can also signal via protein kinases A and C, mitogen-activated protein kinases, and cyclooxygenase 2 pathways, among others.1,3 The CB1 receptor is widely distributed in the central nervous system1 and at lower levels in various peripheral tissues (eg, myocardium,4–6 human coronary artery endothelial and smooth muscle cells,7,8 adipose tissue,9,10 and the liver10–12). Initially it was thought that CB2 receptors were expressed only in immune and hematopoietic cells, but recent studies have also established their presence in the myocardium,6 human coronary endothelial and smooth muscle cells,7,8 brain,13 and liver.12 Modulation of the endocannabinoid system (ECS) may be therapeutically exploited in various cardiovascular (CV) disorders ranging from circulatory shock, stroke, atherosclerosis and restenosis, and hypertension, to cirrhotic cardiomyopathy, myocardial infarction, and chronic heart failure.1,14,15

Role of VR1 and CB1 receptors in modelling of cardio-respiratory response to arvanil, an endocannabinoid and vanilloid hybrid, in rats

Cardio-respiratory effects of an intravenous injection of arvanil, a structural “hybrid” between capsaicin and anandamide, were investigated in 40 urethane-chloralose anaesthetized and spontaneously breathing rats. In the group of rats the response to arvanil was checked to establish the appropriate dose of the drug. To analyze the pattern of the cardio-respiratory effects rats were challenged with bolus injection of arvanil (0.8 mg kg − 1 ) into the femoral vein. Administration of the drug evoked, in all tested rats, a significant increase of tidal volume ( V T) and diaphragm activity, hypertension coupled with a fall in respiratory rate ( f). To test the contribution of vanilloid (VR1) and cannabinoid (CB1) receptors to post-arvanil response, administrations of the drug were preceded by nonselective VR1 antagonist ruthenium red, selective VR1 antagonist SB366791 or selective CB1 antagonist AM281. All antagonists eliminated an increase in V T but failed to block the hypertension evoked by arvanil. Ruthenium red as well as SB366791 abolished post-arvanil fall in respiratory rate. The rise of diaphragm activity was totally eliminated by ruthenium red and markedly reduced by SB366791. AM281 blockade of post-arvanil changes in f and diaphragm activity was ineffective. These findings indicated that the post-arvanil rise of V T was mediated by both VR1 and CB1 receptors. Only vanilloid receptors were involved in the increase of diaphragm activity and decrease of respiratory frequency. Hypertensive response to arvanil might depend on different types of receptors.

TRPV1 Gene Required for Thermosensory Transduction and Anticipatory Secretion from Vasopressin Neurons during Hyperthermia

Increases in core body temperature promote thermoregulatory cooling by stimulating sweat production and preemptive renal water reabsorption through the release of vasopressin (VP, antidiuretic hormone). The mechanism by which the hypothalamus orchestrates this anticipatory VP release during hyperthermia is unknown but has been linked to a central thermosensory mechanism. Here, we report that thermal stimuli spanning core body temperatures activate a calcium-permeable, ruthenium red- and SB366791-sensitive nonselective cation conductance in hypothalamic VP neurons. This response is associated with a depolarizing receptor potential and an increase in action potential firing rate, indicating that these neurons are intrinsically thermosensitive. The thermosensitivity of VP neurons isolated from trpv1 knockout (Trpv1(-/-)) mice was significantly lower than that of wild-type counterparts. Moreover, Trpv1(-/-) mice showed an impaired VP response to hyperthermia in vivo. Channels encoded by the trpv1 gene thus confer thermosensitivity in central VP neurons and contribute to the thermal control of VP release in vivo.

Anandamide inhibits cholangiocyte hyperplastic proliferation via activation of thioredoxin 1/redox factor 1 and AP-1 activation

The endocannabinoid system regulates various aspects of hepatic fibrosis; however, nothing is known about its role in regulating cholangiocyte proliferation and function. We evaluated the effects of anandamide (AEA) on cholangiocyte proliferation and explored the effects of AEA on the thioredoxin 1 (TRX1)/redox factor 1 (Ref1)/activator protein-1 (AP-1) pathway. Mice underwent bile duct ligation (BDL) and were infused with AEA for 3 days postsurgery. Proliferation and apoptosis were evaluated in liver sections. Effects of in vitro AEA treatment on cholangiocyte proliferation and apoptosis were studied in purified cholangiocytes. The relative expression of cannabinoid receptors was also assessed in liver sections and cholangiocytes. mRNA expression of the cannabinoid receptors Cb1 and VR1 was decreased after BDL, whereas there was an upregulation of Cb2 mRNA. AEA decreased cholangiocyte growth and induced accumulation of reactive oxygen species, upregulation of TRX1, Ref1, c-Fos, and c-Jun expression, increased nuclear localization of TRX1, and increased AP-1 transcriptional activity. Specific knockdown of TRX1 or Ref1 expression ablated the AP-1 transcriptional activity and AEA-induced cell death but not expression of c-Fos and c-Jun. Knockdown of c-Fos and c-Jun expression also ablated AEA-induced apoptosis. We conclude that AEA suppresses cholangiocyte proliferation during cholestasis via a Cb2-dependent mechanism. Modulation of the endocannabinoid system may be important in the treatment of cholangiopathies.

Protease-Activated Receptor-2 Augments Experimental Crescentic Glomerulonephritis

Protease-activated receptor-2 (PAR-2) is a cellular receptor expressed prominently on epithelial, mesangial, and endothelial cells in the kidney and on macrophages. PAR-2 is activated by serine proteases such as trypsin, tryptase, and coagulation factors VIIa and Xa. It induces pleiotropic effects including vasodilatation, increasing plasminogen activator inhibitor (PAI-1) expression, mesangial cell proliferation, and cytokine production by macrophages. The role of PAR-2 in renal inflammation was studied in antiglomerular basement membrane antibody-induced crescentic glomerulonephritis (CGN) using PAR-2-deficient (PAR-2(-/-)) mice and wild-type littermate controls. PAR-2(-/-) mice had reduced crescent formation, proteinuria, and serum creatinine compared with wild-type mice 21 days after initiation of CGN. Glomerular accumulation of CD4(+) T cells and macrophages and the number of proliferating cells in glomeruli were similar in both groups. Glomerular fibrin deposition was significantly reduced in PAR-2(-/-) mice, and this was associated with reduced renal plasminogen activator inhibitor expression and increased renal matrix-metalloprotinase-9 activity. These results demonstrate a proinflammatory role for PAR-2 in CGN that is independent of effects on glomerular leukocyte recruitment and mesangial cell proliferation. PAR-2-mediated augmentation of renal plasminogen activator inhibitor expression and inhibition of matrix-metalloprotinase-9 activity may contribute to increased glomerular fibrin accumulation and glomerular injury in CGN.