Laurent, C. et al. (a,b) The A2AR selective agonist CGS21680, 30 nM, has an effect on basal fEPSP slope in Tg(CaMKII-hA2AR) but not in WT animals ; Results were analyzed using a unpaired Student’s t-test (*P < 0.05 comparing to WT); (c) A2AR tonically increase excitatory transmission in Tg(CaMKII-hA2AR) animals, an effect revealed by the inhibitory effect of the A2AR selective antagonist SCH58261 (50 nM) on basal synaptic transmission, that was not observed in WT animals. 10–14 weeks old WT and tg(CaMKII-hA2AR) rats treated either with vehicle or KW 6002, were first handled for 5 days before testing in the behavior assays. PER1, being the only clock gene in the group, exhibits a robust amplitude of rhythmic expression, initiated in the SCN and PVN45. *P < 0.05 compared to control, #P < 0.05 compared with dexamethasone 20 min, ϕP < 0.05 compared with dexamethasone 60 min. Likewise, the potency of HE-NECA to inhibit platelet aggregation at 12 and 60 hours after caffeine withdrawal was significantly increased. For further in vitro experiments, platelet-rich plasma (PRP) from 3 additional subjects was incubated at 37°C in a thermostatic bath for 6 or 12 hours in the absence or presence of caffeine (20 μmol/L) or SCH 58261 (100 nmol/L). Herman, J. P. & Spencer, R. Regulation of hippocampal glucocorticoid receptor gene transcription and protein expression in vivo . 1a). Drug treatments were performed at day 8, 1 h after replacing the medium by Neurobasal without B27. 4G), vGluT1 (Fig. The IC50 values for HE-NECA to inhibit ADP-induced platelet aggregation were 50±5 and 30±2 nmol/L at 12 and 60 hours after caffeine withdrawal, respectively. CUS does not significantly affect spontaneous locomotion. *P < 0.05 vs. time exploring the nondisplaced object; Student’s t test. Acta physiologica 199, 161–169, 10.1111/j.1748-1716.2010.02115.x (2010). The present report shows that A2AR not only regulate HPA-axis function, but also directly modulate GR, which represent key findings for understanding the mechanisms by which A2AR antagonism is effective. The only molecular targets for caffeine at nontoxic doses are the main adenosine receptors in the brain, namely the inhibitory A1 receptors (A1R) and the facilitatory A2A receptors (A2AR) (9). *P < 0.05 compared to control, ΦP < 0.05 compared with dexamethasone induced luciferase activity calculated using a one-way ANOVA followed by a Bonferroni post hoc test. Background—Caffeine acts mainly via blockade of adenosine receptors, which have been classified into A1, A2A, A2B, and A3 subtypes. (B) Caffeine reaches similar plasma concentrations in control mice and in mice subjected to CUS, as measured by HPLC. Biol Psychiatry 74, 357–366, 10.1016/j.biopsych.2012.12.003 (2013). The A2AR selective antagonist 2-(2-furanyl)-7-(2-phenylethyl)-7H-pyrazolo[4,3-e][1,2,4]triazolo[1,5-c]pyrimidin-5-amine (SCH58261), the A2AR selective agonist 4-[2-[[6-amino-9-(N-ethyl-β-D-ribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzene propanoic acid (CGS21680) and the GR antagonist (11β,17β)-11-[4-(dimethylamino)phenyl]-17-hydroxy-17-(1-propynyl)-estra-4,9-dien-3-one (RU486) were purchased from Tocris Cookson, UK. CTR, control. Nat Neurosci 1, 69–73, 10.1038/271 (1998). 2d). In contrast, CUS enhanced the A2AR antagonist [3H]5-amino-7-2-phenylethyl.-2-2-furyl-pyrazolo[4,3-ex-1,2,4-triazolo-1,5]pyrimidine ([3H]SCH58261) binding in the synaptosomal membranes (which was 60.1 ± 8.7% larger in CUS-treated mice than in control mice; n = 6, P < 0.05) but not in total or in gliosomal membranes of the hippocampus (Fig. Both the number of crossings and the number of rearing events were globally similar in nonstressed control mice (ctr) and in mice subjected to CUS. 1F) (n = 36–39 mice per group). n = 5; *P < 0.05, two-way ANOVA [CUS F(1,16) = 19.15; genotype F(1,16) = 10.13; interaction F(1,16) = 4.16] followed by a Newman–Keuls post hoc test. mRNA levels of GILZ (a), PER1 (b) and Bcl2 (c) increase in primary neurons after incubation with dexamethasone (100 nM) for 1 h an effect prevented by the presence of the selective A2AR antagonist SCH 51280 (50 nM), n = 3–5. Data are shown as mean ± SEM of 5 or 6 mice per group; *P < 0.05 using an unpaired Student’s t test. Binding Parameters of the A2A Adenosine Receptor Antagonist [3H]SCH 58261 in Platelet Membranes and Potency of the A2A Adenosine Receptor Agonist HE-NECA to Increase Platelet cAMP and to Inhibit Platelet Aggregation. PLoS One 6, e21660, 10.1371/journal.pone.0021660 (2011). and V.L.B. However, the molecular targets operated by caffeine to afford these beneficial effects have not been defined. 1d), which is comparable to levels found in physiological aging27. In fact, in dogs, adenosine released during myocardial ischemia inhibits platelet aggregation, an effect that is antagonized by 8-phenyltheophylline.9 If one assumes that endogenous released adenosine during episodes of ischemia inhibits platelet aggregation and thromboembolization, it is conceivable that mild chronic caffeine consumption may lead to a paradoxical reduction in platelet aggregability.

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