Department of Systems Medicine, Tor Vergata University of Rome, 00133 Rome, Italy.
Department of Biology and Biotechnology "Charles Darwin", Sapienza University of Rome, 00185 Rome, Italy.
Int J Mol Sci. 2021 Mar 7;22(5):2691. doi: 10.3390/ijms22052691.
We aimed to investigate A2A receptors in the basal ganglia of a DYT1 mouse model of dystonia. A2A was studied in control Tor1a+/+ and Tor1a+/- knock-out mice. A2A expression was assessed by anti-A2A antibody immunofluorescence and Western blotting. The co-localization of A2A was studied in striatal cholinergic interneurons identified by anti-choline-acetyltransferase (ChAT) antibody. A2A mRNA and cyclic adenosine monophosphate (cAMP) contents were also assessed. In Tor1a+/+, Western blotting detected an A2A 45 kDa band, which was stronger in the striatum and the globus pallidus than in the entopeduncular nucleus. Moreover, in Tor1a+/+, immunofluorescence showed A2A roundish aggregates, 0.3-0.4 μm in diameter, denser in the neuropil of the striatum and the globus pallidus than in the entopeduncular nucleus. In Tor1a+/-, A2A Western blotting expression and immunofluorescence aggregates appeared either increased in the striatum and the globus pallidus, or reduced in the entopeduncular nucleus. Moreover, in Tor1a+/-, A2A aggregates appeared increased in number on ChAT positive interneurons compared to Tor1a+/+. Finally, in Tor1a+/-, an increased content of cAMP signal was detected in the striatum, while significant levels of A2A mRNA were neo-expressed in the globus pallidus. In Tor1a+/-, opposite changes of A2A receptors' expression in the striatal-pallidal complex and the entopeduncular nucleus suggest that the pathophysiology of dystonia is critically dependent on a composite functional imbalance of the indirect over the direct pathway in basal ganglia.
我们旨在研究多巴反应性肌张力障碍 1 型(DYT1)小鼠模型基底神经节中的 A2A 受体。在对照 Tor1a+/+和 Tor1a+/-敲除小鼠中研究 A2A。通过抗 A2A 抗体免疫荧光和 Western blot 评估 A2A 的表达。通过抗胆碱乙酰转移酶(ChAT)抗体鉴定纹状体胆碱能中间神经元来研究 A2A 的共定位。还评估了 A2A mRNA 和环磷酸腺苷(cAMP)含量。在 Tor1a+/+中,Western blot 检测到 A2A 45 kDa 带,在纹状体和苍白球中的表达强于在动眼神经核。此外,在 Tor1a+/+中,免疫荧光显示 A2A 呈圆形聚集体,直径为 0.3-0.4 μm,在纹状体和苍白球的神经丛中比在动眼神经核中更密集。在 Tor1a+/-中,A2A Western blot 表达和免疫荧光聚集体在纹状体和苍白球中增加,或在动眼神经核中减少。此外,在 Tor1a+/-中,与 Tor1a+/+相比,A2A 聚集体在 ChAT 阳性中间神经元上的数量增加。最后,在 Tor1a+/-中,在纹状体中检测到 cAMP 信号的含量增加,而在苍白球中则新表达了显著水平的 A2A mRNA。在 Tor1a+/-中,纹状体-苍白球复合体和动眼神经核中 A2A 受体表达的相反变化表明,肌张力障碍的病理生理学严重依赖于基底神经节中间接通路对直接通路的复合功能失衡。