Department of Systems Medicine, Tor Vergata University of Rome, via Montpellier 1, 00133 Rome, Italy.
Santa Lucia Foundation, via del Fosso di Fiorano 64, 00143 Rome, Italy.
Int J Mol Sci. 2020 Feb 6;21(3):1073. doi: 10.3390/ijms21031073.
Dystonia pathophysiology has been partly linked to downregulation and dysfunction ofdopamine D2 receptors in striatum. We aimed to investigate the possible morpho-structuralcorrelates of D2 receptor downregulation in the striatum of a DYT1 Tor1a mouse model. Adultcontrol Tor1a+/+ and mutant Tor1a+/- mice were used. The brains were perfused and free-floatingsections of basal ganglia were incubated with polyclonal anti-D2 antibody, followed by secondaryimmune-fluorescent antibody. Confocal microscopy was used to detect immune-fluorescent signals.The same primary antibody was used to evaluate D2 receptor expression by western blot. The D2receptor immune-fluorescence appeared circumscribed in small disks (0.3-0.5 μm diameter), likelyrepresenting D2 synapse aggregates, densely distributed in the striatum of Tor1a+/+ mice. In theTor1a+/- mice the D2 aggregates were significantly smaller (μm2 2.4 ± SE 0.16, compared to μm2 6.73± SE 3.41 in Tor1a+/+) and sparse, with ~30% less number per microscopic field, value correspondentto the amount of reduced D2 expression in western blotting analysis. In DYT1 mutant mice thesparse and small D2 synapses in the striatum may be insufficient to "gate" the amount ofpresynaptic dopamine release diffusing in peri-synaptic space, and this consequently may result ina timing and spatially larger nonselective sphere of influence of dopamine action.
Dystonia 的病理生理学部分与纹状体中多巴胺 D2 受体的下调和功能障碍有关。我们旨在研究 DYT1 Tor1a 小鼠模型纹状体中 D2 受体下调的可能形态结构相关性。使用成年对照 Tor1a+/+ 和突变 Tor1a+/- 小鼠。对大脑进行灌流,并将基底神经节的游离切片与多克隆抗 D2 抗体孵育,然后用二级免疫荧光抗体孵育。使用共聚焦显微镜检测免疫荧光信号。用相同的一抗通过 Western blot 评估 D2 受体表达。D2 受体免疫荧光呈小圆盘状(直径约 0.3-0.5μm),可能代表 D2 突触聚集物,在 Tor1a+/+ 小鼠的纹状体中密集分布。在 Tor1a+/- 小鼠中,D2 聚集物明显更小(μm2 2.4±SE 0.16,与 Tor1a+/+ 的 μm2 6.73±SE 3.41 相比)且稀疏,每个显微镜视野的数量减少了约 30%,这与 Western blot 分析中 D2 表达减少的量相对应。在 DYT1 突变小鼠中,纹状体中稀疏和小的 D2 突触可能不足以“控制”扩散到突触间隙中的突触前多巴胺释放量,因此这可能导致多巴胺作用的非选择性影响范围在时间和空间上更大。