Laboratório de Neuroquímica e Psicofarmacologia, Escola de Ciências da Saúde e da Vida, Pontificia Universidade, Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre, Brazil.
Programa de Pós-Graduação me Medician e Ciências da Saúde, Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre, Brazil.
Zebrafish. 2023 Oct;20(5):210-220. doi: 10.1089/zeb.2023.0002.
The A1 adenosine receptor is the most widely expressed P1 receptor in vertebrates, performing inhibitory tone of the nervous system. Increased levels of adenosine are crucial to promote tissue protection in threatening situations, such as convulsion and hypoxia. Zebrafish is an established model organism for studies on health and disease. In this study, we evaluated the functionality of A adenosine receptor through development of zebrafish (6-7-day-, 3-, 8-, and 24-month-old), assessing: (I) the effects of the agonist N-cyclopenthyladenosine (CPA) over locomotor parameters, (II) the anticonvulsant properties of CPA and adenosine in the pentylenetetrazol-induced seizure, and (III) the gene expression of through development. CPA promoted decreased distance traveled in the highest concentrations/doses tested (larvae: 75 to 500 μM; adults: 20 mg.kg), altered mean velocity (larvae: 50-500 μM; adults: 20 mg.kg) and time in the bottom zone of apparatus (adults: decrease in 20 mg.kg). Adenosine increased the latency of the larvae to reach stage II at 5 and 10 μM. CPA anticonvulsant effect against convulsive stage II was reached at 75 μM, although it decreased basal locomotor activity in larvae. For adults, CPA 10 mg.kg was effective as anticonvulsant without locomotory effects. Adenosine had minor anticonvulsant effects in the concentration tested (larvae: 5 and 10 μM). The level of gene expression of was stable in brain from adult animals (8- and 24-month-old animals). These results suggest that zebrafish has similar responses to CPA as mammals. To avoid confounding factors, such as locomotor effects, during any brain function investigation using A adenosine receptor as a target, the concentration below 75 μM or below the dose of 20 mg.kg of CPA is ideal for zebrafish at larval and adult stages, respectively.
A1 腺苷受体是脊椎动物中表达最广泛的 P1 受体，对神经系统起抑制作用。在威胁情况下，如惊厥和缺氧，腺苷水平的升高对于促进组织保护至关重要。斑马鱼是研究健康和疾病的成熟模式生物。在这项研究中，我们通过发育中的斑马鱼（6-7 天、3 个月、8 个月和 24 个月）评估了 A 腺苷受体的功能，评估了：（I）激动剂 N-环戊基腺苷（CPA）对运动参数的影响，（II）CPA 和腺苷在戊四氮诱导的惊厥中的抗惊厥作用，以及（III）通过发育的表达。CPA 在测试的最高浓度/剂量下促进了运动距离的减少（幼虫：75 至 500μM；成鱼：20mg/kg），改变了平均速度（幼虫：50-500μM；成鱼：20mg/kg）和仪器底部区域的时间（成鱼：20mg/kg 减少）。腺苷增加了幼虫达到 5 和 10μM 时达到 II 期的潜伏期。CPA 对抗惊厥 II 期的抗惊厥作用在 75μM 时达到，但在幼虫中降低了基础运动活性。对于成鱼，CPA 10mg/kg 作为抗惊厥剂是有效的，没有运动效果。腺苷在测试浓度下具有较小的抗惊厥作用（幼虫：5 和 10μM）。成年动物（8 个月和 24 个月大的动物）大脑中表达的基因水平稳定。这些结果表明，斑马鱼对 CPA 的反应与哺乳动物相似。为了避免使用 A 腺苷受体作为靶点进行任何脑功能研究时出现运动效应等混杂因素，在幼虫和成年阶段，CPA 的浓度低于 75μM 或低于 20mg/kg 的剂量是理想的。
