Blockade of Adenosine A2A and ATP P2Y1 Receptors Prevents ILIB Associated Neuronal Toxicity

Abstract

Event Abstract Back to Event Blockade of Adenosine A2A and ATP P2Y1 Receptors Prevents ILIB Associated Neuronal Toxicity Ana Patricia-Simoes1*, Carla G. Silva1, Joao Andre Duarte1, Paula M. Canas1 and Rodrigo A. Cunha1 1 University of Coimbra, Center for Neuroscience of Coimbra, Institute of Biochemistry, Portugal Interleukin-1beta (IL1β) and tumor necrosis factor-α (TNF-α) are pro-inflammatory cytokines known to aggravate neuronal damage caused by different insults. This effect is achieved through cytokine-induced activation (phosphorylation) of stress kinases such as JNK and p38. We have previously shown that A2A or P2Y1 receptors (A2AR and P2Y1R) blockade affords neuroprotection under several neurotoxic conditions. Moreover, A2AR blockade prevents both neuroinflammation-induced increases of IL1β levels and consequent depression of long-term potentiation. Here, we investigated if A2AR and P2Y1R can also control inflammation-induced exacerbation of neuronal damage.Hippocampal neuronal cultures, 7-8 DIV, were exposed to IL1β (1, 10, 100 ng/ml) or to TNF-α (1, 5 ng/ml) for different times. JNK and p38 phosphorylation levels were analysed by Western blot and immunocytochemistry (ICC). These cytokines effect on glutamate-mediated excitotoxicity was studied by adding glutamate (100 μM) together with IL-1β (100 ng/ml) or TNF-α (5 ng/ml) in the cell culture medium, during 25 min, and evaluating cell viability 24 hrs later by syto-13/propidium iodide (PI) staining and lactate dehydrogenase (LDH) assay. Western blot and ICC analysis revealed that either IL1β (100 ng/ml; 15 min) or TNF-α (5 ng/ml; 10 min) activated p38 (IL1β: 189±14%, n=8, p<0.05; TNF-α: 385±51%, n=9, p<0.05) and that IL1β also activated JNK (138±9%, n=5, p<0.01). Blockade of A2AR, with the antagonist SCH58261 (50 nM), or P2Y1R, with antagonist MRS2179 (10 μM), prevented and attenuated, respectively, IL1β-induced activation of p38 (A2AR: 108.0±20%, n=5, p<0.05; P2Y1R: 148±19%, n=7) and prevented IL1β-induced activation of JNK (A2AR: 74±8%, n=5, p<0.01; P2Y1R: 82±6%, n=4, p<0.01). However, neither was able to prevent TNF-α-induced p38 activation. Moreover, Syto-13 and PI staining showed that cell viability was reduced by glutamate (21±3%, n=6), which was exacerbated by IL1β (51±14%, n=7). Blockade of A2AR or P2Y1R diminishes the exacerbation of glutamate toxicity by IL1β (17±9% and 22±18% of nonviable cells, n=3-4, respectively). Likewise, IL1β and glutamate induced LDH leakage (160±29% of control, n=3) which was attenuated by either A2AR or P2Y1R antagonist (97±25% and 117±16% of control, n=3, respectively). Values are mean±SEM of n experiments.These results suggest that the neuroprotection afforded by blockade of A2ARs or P2Y1Rs might also involve the particular control of IL1β direct effects on neurons. Conference: 11th Meeting of the Portuguese Society for Neuroscience, Braga, Portugal, 4 Jun - 6 Jun, 2009. Presentation Type: Poster Presentation Topic: Neurodegenerative Disorders Citation: Patricia-Simoes A, Silva CG, Andre Duarte J, Canas PM and Cunha RA (2009). Blockade of Adenosine A2A and ATP P2Y1 Receptors Prevents ILIB Associated Neuronal Toxicity. Front. Neurosci. Conference Abstract: 11th Meeting of the Portuguese Society for Neuroscience. doi: 10.3389/conf.neuro.01.2009.11.090 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: 10 Aug 2009; Published Online: 10 Aug 2009. * Correspondence: Ana Patricia-Simoes, University of Coimbra, Center for Neuroscience of Coimbra, Institute of Biochemistry, Alicante, Portugal, anapatriciafrsimoes@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 Ana Patricia-Simoes Carla G Silva Joao Andre Duarte Paula M Canas Rodrigo A Cunha Google Ana Patricia-Simoes Carla G Silva Joao Andre Duarte Paula M Canas Rodrigo A Cunha Google Scholar Ana Patricia-Simoes Carla G Silva Joao Andre Duarte Paula M Canas Rodrigo A Cunha PubMed Ana Patricia-Simoes Carla G Silva Joao Andre Duarte Paula M Canas Rodrigo A Cunha 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.