Role Of Adenosine A2A Receptors Research Articles

Targeted deletion of A2A adenosine receptors attenuates the protective effects of myocardial postconditioning

Endogenous adenosine is an important ligand trigger for the cardioprotective effects of postconditioning (POC), yet it is unclear which adenosine receptor subtype is primarily responsible. To evaluate the role of A(2A) adenosine receptors in POC-induced protection, global ischemia-reperfusion was performed with and without POC in isolated wild-type (WT) and A(2A) adenosine receptor knockout (A(2A)KO) mouse hearts. Injury was measured in terms of postischemic functional recovery and release of cardiac troponin I (cTnI). Activation of protective signaling with POC was assessed by Akt and extracellular signal-regulated kinase (ERK) 1/2 phosphorylation. In WT hearts, POC improved recovery of postischemic developed pressure in early (81.6 +/- 6.4% of preischemic baseline vs. 37.5 +/- 5.6% for non-POC WT at 1 min) and late (62.2 +/- 4.2% of baseline vs. 45.5 +/- 5.3% for non-POC WT at 30 min) reperfusion, reduced cTnI release by 37%, and doubled the phosphorylation of both Akt and ERK1/2. These beneficial effects of POC were blocked by treatment with the selective A(2A) adenosine receptor antagonist ZM-241385 during reperfusion. Postischemic functional recovery, cTnI release, and phosphorylation of Akt and ERK1/2 were not different between non-POC WT and A(2A)KO hearts. In A(2A)KO hearts, POC did not improve functional recovery, reduce cTnI release, nor increase phosphorylation of Akt or ERK1/2. Thus the protective effects of POC are attenuated by both selective A(2A) receptor antagonism and targeted deletion of the gene encoding A(2A) adenosine receptors. These observations support the conclusion that endogenous activation of A(2A) adenosine receptors is an essential trigger leading to the protective effects of POC in isolated murine hearts.

A Genetic Variation in the Adenosine A2A Receptor Gene (ADORA2A) Contributes to Individual Sensitivity to Caffeine Effects on Sleep

Caffeine is the most widely used stimulant in Western countries. Some people voluntarily reduce caffeine consumption because it impairs the quality of their sleep. Studies in mice revealed that the disruption of sleep after caffeine is mediated by blockade of adenosine A2A receptors. Here we show in humans that (1) habitual caffeine consumption is associated with reduced sleep quality in self-rated caffeine-sensitive individuals, but not in caffeine-insensitive individuals; (2) the distribution of distinct c.1083T>C genotypes of the adenosine A2A receptor gene (ADORA2A) differs between caffeine-sensitive and -insensitive adults; and (3) the ADORA2A c.1083T>C genotype determines how closely the caffeine-induced changes in brain electrical activity during sleep resemble the alterations observed in patients with insomnia. These data demonstrate a role of adenosine A2A receptors for sleep in humans, and suggest that a common variation in ADORA2A contributes to subjective and objective responses to caffeine on sleep.

Effects of adenosine receptor antagonists on amitriptyline-induced vasodilation in rat isolated aorta

Background. Although we have previously demonstrated the beneficial effects of adenosine receptor antagonists in preventing cardiovascular toxicity of amitriptyline in rats, it is not clear whether adenosine receptors in heart or in vasculature are dominant. The aim of the current study was to investigate the role of adenosine A2a receptors on amitriptyline-induced vasodilation in rat isolated aorta. Methods. After determining EC80 of noradrenalin (NA) (the concentration of noradrenalin that produces 80% of maximal contractile response) as 10−5M, the IC50 value of amitriptyline was measured in rat isolated aorta (the drug concentration causing a half- maximal inhibition of contractile responses to NA); IC50 of amitriptyline was then compared in the presence of the DPCPX (a selective adenosine A1 antagonist), CSC (a selective A2a antagonist) or DMSO (a solvent for adenosine antagonists). Statistical analysis was done using the Student t test. Results. Amitriptyline-inhibited 49.9 ± 3.7 % contractile response to NA on aorta segments at 1.8 × 10−5M (IC50). While DPCPX increased amitriptyline-induced inhibition on contractile response to NA dose dependently, CSC decreased the contractile response to NA only at 10−5M. DMSO did not change amitriptyline-induced IC50. Conclusion. Adenosine A2a receptor stimulation seems to be responsible partly for amitriptyline-induced vasodilation and hypotension since the adenosine A1 antagonist, DPCPX, increased amitriptyline-induced vasodilation in rat isolated aorta.

Attenuation of nicotine-induced rewarding effects in A 2A knockout mice

The non-selective A 2A antagonist caffeine has been reported to modify nicotine-induced locomotor and reinforcing effects. In the present study, we have investigated the specific role of A 2A adenosine receptors in the behavioural responses induced by nicotine by using genetically modified mice lacking A 2A adenosine receptors. Acute nicotine administration induced a similar decrease of locomotor activity in A 2A knockout mice and wild-type littermates. Acute antinociceptive responses elicited by nicotine in the tail-immersion and hot-plate tests were unaffected in these mutant mice. The rewarding properties of nicotine were then investigated using the place-conditioning paradigm. Nicotine-induced conditioned place preference was suppressed in A 2A knockout mice. Accordingly, in vivo microdialysis studies revealed that the extracellular levels of dopamine in the nucleus accumbens were not increased after nicotine administration in mutant mice. Wild-type and A 2A knockout mice were trained in conditioned taste aversion procedure in which drinking a saccharin or saline solution was paired with nicotine or saline injections. A similar reduction in the intake of nicotine-paired solution in this paradigm was obtained in both genotypes. Finally, the administration of the nicotinic antagonist mecamylamine in nicotine-dependent mice precipitated a similar withdrawal syndrome in both genotypes. Together, the present results identify A 2A adenosine receptors as an important factor that contributes to the rewarding properties of nicotine.

Increased oxidative stress and depletion of antioxidants in A 2A adenosine receptor deficient allergic mice

Adenosine has multiple roles through its specific cell surface receptors leading to various physiological responses. A2A receptor activation possesses potent anti-inflammatory properties, which may also lead to antioxidant function. The present study was undertaken to investigate the role of adenosine A2A receptor on oxidant-antioxidant balance in lung and systemic circulation in an allergic asthma model in adenosine A2A receptor knock-out (KO) mice. A2A KO and wild-type (WT) mice were sensitized with two i.p. injections of ragweed (200 μg) on days 1 and 6. Aerosolization challenge was performed with 0.5% ragweed for 20 minutes both in the morning and afternoon on days 11, 12 and 13. Twenty-four hrs after the last challenge, bronchoalveolar lavage fluid (BALF) and blood was collected from these mice for various assays. Allergen challenge in sensitized mice increased malondialdehyde-thiobarbituric acid adducts, both in plasma (p<0.05) and BALF (p<0.01) of WT and KO, with KO having greater oxidative stress in BALF than WT. Moreover, plasma total nitrates and nitrites as a measure of nitric oxide production were also found to be higher in A2A KO allergen challenged mice as compared to their WT and controls (p<0.01). In contrast, total antioxidants in plasma (p<0.05) were depleted in the KO mice, while BALF having greater depletion (p<0.01) both in WT and KO mice. These data showed that A2A KO mice had increased oxidative stress and depletion of antioxidants after allergen challenge both in the lung and systemic circulation, suggesting possible antioxidant function of A2A receptor in this model of asthma. (Supported by HL-027339)

A2a receptors mediate inhibitory effects of adenosine on colonic motility in the presence of experimental colitis

Adenosine regulates immunity and inflammation, and acts also as a modulator of gut functions. In this study, we investigated the role of adenosine A2a receptors on colonic motility in a rat model of experimental colitis. Colitis was induced by 2,4-dinitrobenzenesulfonic acid. The effects of ZM 241385 (A2a receptor antagonist) and CGS 21680 (A2a receptor agonist) were assayed on cholinergic contractions of colonic longitudinal muscle preparations evoked by transmural electrical stimulation (TES) or carbachol. A2a receptor expression in colonic neuromuscular layers was assessed by reverse transcription-polymerase chain reaction. ZM 241385 increased TES-induced contractions in the absence or in the presence of colitis, the drug being more effective in colonic preparations from inflamed animals. The enhancing effects of ZM 241385 were unaffected by guanethidine or alpha-chimotrypsin, whereas being prevented by Nomega-propyl-L-arginine (neuronal nitric oxide synthase inhibitor) or adenosine 5'-(alpha,beta-methylene) diphosphate (ecto-5'-nucleotidase inhibitor). Upon exposure of colonic tissues from normal or inflamed rats to dipyridamole plus adenosine deaminase, to abate endogenous adenosine levels, CGS 21680 evoked concentration-dependent reductions of contractile responses to TES, which were more intense in preparations from inflamed rats, and were antagonized by ZM 241385. Neither CGS 21680 nor ZM 241385 affected carbachol-induced contractions. Reverse transcription-polymerase chain reaction showed an increase in A2a receptor expression in colonic tissues isolated from inflamed animals. The adenosine system is involved in neuroplastic changes occurring in inflamed gut. A2a receptors modulate the activity of colonic excitatory cholinergic nerves via facilitatory control on inhibitory nitrergic pathways, and such a regulatory function is enhanced in the presence of bowel inflammation.

The Lack of A2A Adenosine Receptors Diminishes the Reinforcing Efficacy of Cocaine

Adenosine is an endogenous purine nucleoside, which acts as a neuromodulator in the central nervous system. A2A adenosine and D2 dopamine receptors are colocalized in the same neurons in discrete brain areas, and the dopaminergic transmission plays a crucial role in the addictive properties of drugs of abuse, such as cocaine. In the present study, we have investigated the specific role of A2A adenosine receptors in cocaine-induced behavioral responses related to its addictive properties. For this purpose, we have evaluated the acute locomotor effects produced by cocaine and the development of locomotor sensitization by repeated cocaine administration. In addition, we have also examined cocaine acute rewarding properties using the conditioned place preference. Finally, we used the intravenous drug self-administration paradigm to investigate the acquisition of an operant response maintained by cocaine self-administration and the reinforcing efficacy of the drug in these knockout animals. Acute cocaine induced a similar increase of locomotor activity in mice lacking A2A adenosine receptors and wild-type littermates. Cocaine-induced locomotor sensitization and conditioned place preference were also maintained in A2A knockout mice. Nevertheless, these knockout mice showed a lower rate of cocaine self-administration than wild-type mice in both fixed ratio 1 and 3 schedules of reinforcement. Moreover, a reduction in the maximal effort to obtain a cocaine infusion was found in A2A knockout mice under a progressive ratio schedule. In addition, a vertical shift of the cocaine dose-response curve was observed in mice lacking A2A adenosine receptors in comparison with wild-type littermates. Our study demonstrates that A2A adenosine receptors play an important role in cocaine addictive properties, and these receptors seem to be required to develop the addictive effects of this drug.

New aspects of physiological and pathophysiological functions of adenosine A2A receptor in basal ganglia.

There is now growing interest in the functional role of adenosine A2A receptors. Their distribution within the brain is restricted in the basal ganglia, particularly abundant in the striatum, which are thought to play a crucial role in the control of motor behavior. Indeed, newly developed A2A receptor selective antagonists have a profound influence on motor functions, with anti-Parkinsonian activities in several animal models. Striatal spiny neurons serve as a major anatomical locus for the relay of cortical information flow through the basal ganglia. The GABA releasing projection neurons represent the A2A receptor-mediated main target of adenosine. The GABAergic synaptic neurotransmission is regulated by adenosine via A2A receptors on the presynaptic terminals. Blockade of this modulatory function by A2A antagonists could repair striatopallidal abnormal neuronal activities provoked by striatal dopamine depletion in the Parkinsonian state. A2A receptor antagonists provide a novel therapeutic potential for the treatment of Parkinson's disease.

Electrophysiological and behavioural evidence for an antagonistic modulatory role of adenosine A2A receptors in dopamine D2 receptor regulation in the rat dopamine-denervated striatum.

It has been shown that striatal adenosine A2A receptors can antagonistically interact with dopamine D2 receptors at the membrane level leading to a decrease in the affinity and efficacy of D2 receptors. Extracellular recordings and rotational behaviour were employed to obtain a correlate to these findings in an animal model of Parkinson's disease (PD). The recordings were performed in rats with unilateral 6-hydroxydopamine (6-OHDA)-induced catecholamine depletion. While recording in the dopamine-depleted striatum, local applications of the dopamine D2 agonist quinpirole reduced neuronal activity. However, when the adenosine A2A antagonist MSX-3 was applied simultaneously with quinpirole, the inhibition of neuronal firing seen after quinpirole alone was significantly potentiated (P< 0.001, n = 11). In contrast, local application of CGS 21680 attenuated the effect of quinpirole. The doses of MSX-3 and CGS 21680 used to achieve the modulation of quinpirole action had no effect per se on striatal neuronal firing. Furthermore, rotational behaviour revealed that MSX-3 dose-dependently increased the number of turns when administrated together with a threshold dose of quinpirole while no enhancement was achieved when MSX-3 was combined with SKF 38393. MSX-3 alone did not induce rotational behaviour. In conclusion, this study shows that low ineffective doses of MSX-3 enhance the effect of quinpirole on striatal firing rate, while the A2A agonist exerts the opposite action. This mechanism gives a therapeutic potential to A2A antagonists in the treatment of PD by enhancing D2 receptor function.

Effect of A2A adenosine receptor stimulation and antagonism on synaptic depression induced by in vitro ischaemia in rat hippocampal slices.

1. In the present study we investigated the role of A2A adenosine receptors in hippocampal synaptic transmission under in vitro ischaemia-like conditions. 2. The effects of adenosine, of the selective A2A receptor agonist, CGS 21680 (2-[p-(2-carboxyethyl)-phenethylamino]-5'-N-ethylcarboxamidoade nos ine ), and of selective A2A receptor antagonists, ZM 241385 (4-(2-[7-amino-2-(2-furyl)-¿1,2,4¿-triazolo¿2,3-a¿¿1,3, 5¿triazin-5-ylamino]ethyl)phenol) and SCH 58261 (7-(2-phenylethyl)-5-amino-2-(2-furyl)-pyrazolo-[4,3-e]-1,2, 4-triazolo[1,5-c]pyrimidine), have been evaluated on the depression of field e.p.s.ps induced by an in vitro ischaemic episode. 3. The application of 2 min of in vitro ischaemia brought about a rapid and reversible depression of field e.p.s.ps, which was completely prevented in the presence of the A1 receptor antagonist DPCPX (1, 3-dipropyl-8-cyclopentylxanthine) (100 nM). On the other hand both A2A receptor antagonists, ZM 241385 and SCH 58261, by themselves did not modify the field e.p.s.ps depression induced by in vitro ischaemia. 4. A prolonged application of either adenosine (100 micronM) or CGS 21680 (30, 100 nM) before the in vitro ischaemic episode, significantly reduced the synaptic depression. These effects were antagonized in the presence of ZM 241385 (100 nM). 5. SCH 58261 (1 and 50 nM) did not antagonize the effect of 30 nM CGS 21680 on the ischaemia-induced depression. 6. These results indicate that in the CA1 area of the hippocampus the stimulation of A2A adenosine receptors attenuates the A1-mediated depression of synaptic transmission induced by in vitro ischaemia.

Role of A2A adenosine receptors in inflammation

A2A adenosine receptors are expressed on immune cells including neutrophils, lymphocytes, eosinophils, monocytes/macrophages, and mast cells. Activation of A2A receptors on these cells stimulates an increase in [cyclic AMP]i and causes a diminution of inflammatory responses. In mast cells, degranulation is inhibited; in neutrophils, adherence is reduced and the release of reactive oxygen species is inhibited; in monocytes, differentiation to macrophages is inhibited and the release of tumor necrosis factor-α is inhibited; and in lymphocytes, TCR-triggered interleukin-2 α chain (CD25) up-regulation is reduced. In vivo, selective adenosine A2A agonists decrease inflammation in both infectious and noninfectious models. High concentrations (micromolar) of the A3 selective agonist, IB-MECA, produce anti-inflammatory responses that are mediated by A2A receptors. Selective activation of A2A adenosine receptors with pharmaceutical agents may be a useful strategy for ameliorating an inappropriate and/or an extensive inflammatory response. Drug Dev. Res. 45:103–112, 1998. © 1998 Wiley-Liss, Inc.

A2A adenosine receptors inhibit ATP-induced Ca2+ influx in PC12 cells by involving protein kinase A.

The regulatory role of A2A adenosine receptors in P2 purinoceptor-mediated calcium signaling was investigated in rat pheochromocytoma (PC12) cells. When PC12 cells were treated with 2-p-(2-carboxyethyl)-phenethylamino-5'-N-ethylcarboxamidoadenosine (CGS-21680), a specific agonist of the A2A adenosine receptor, the extracellular ATP-evoked rise in cytosolic free Ca2+ concentration ([Ca2+]i) was inhibited by 20%. Both intracellular calcium release and inositol 1,4,5-trisphosphate production evoked by ATP were not affected by CGS-21680 treatment. However, ATP-evoked Ca2+ influx was inhibited following CGS-21680 stimulation. The CGS-21680-mediated inhibition occurred independently of nifedipine-induced inhibition of the [Ca2+]i rise. The CGS-21680-induced inhibition was completely blocked by reactive blue 2. The CGS-21680 effect was mimicked by forskolin and dibutyryl-cyclic AMP and blocked by Rp-adenosine 3',5'-cyclic monophosphothioate, a protein kinase A inhibitor, or by staurosporine, a general kinase inhibitor. The data suggest that in PC12 cells activation of A2A adenosine receptors leads to inhibition of P2 purinoceptor-mediated Ca2+ influx through ATP-gated cation channels and involves protein kinase A.

The role of adenosine A2a receptors in regulating GABAergic synaptic transmission in striatal medium spiny neurons

The role of adenosine A2a receptors in regulating GABAergic synaptic transmission in striatal medium spiny neurons