Espejo-Porras Francisco, Fernández-Ruiz Javier, de Lago Eva
a Departamento de Bioquímica y Biología Molecular , Instituto Universitario de Investigación en Neuroquímica, Facultad de Medicina, Universidad Complutense , Madrid , Spain.
b Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED) , Madrid , Spain , and.
Amyotroph Lateral Scler Frontotemporal Degener. 2018 Aug;19(5-6):377-386. doi: 10.1080/21678421.2018.1425454. Epub 2018 Jan 15.
We have investigated the endocannabinoid system in the motor cortex of motor neuron disease (MND) patients.
Post-mortem samples from MND patients and controls were used for immunostaining and/or Western blotting analysis of endocannabinoid elements.
We did not find any evidence of neuronal losses in the motor cortex of MND patients, but elevations in glial markers Iba-1 and GFAP were evident. We found no changes in FAAH and MAGL enzymes and in the CB receptor, which correlated with the lack of cortical neuron death. By contrast, the Western blotting analysis of CB receptors proved an increase in the motor cortex corroborated by immunostaining, correlating with the elevated gliosis in these patients. Double-labeling analyses revealed that this elevated CB receptor immunostaining was located in GFAP-labelled astroglial cells. However, we also found CB receptor labeling in cortical neurons confirmed with double immunofluorescence with the neuronal marker MAP-2. This was also found in the spinal cord, using double-labeling with the spinal motor neuron marker choline-acetyl transferase. This happened in both patients and controls, despite these neurons experienced an important degeneration in patients reflected in reduced Nissl staining, TDP-43 immunostaining and CB receptor levels measured by Western blotting.
We have confirmed that CB receptors are elevated in the motor cortex of MND patients associated with the reactive gliosis. This phenomenon is previous to neuronal losses. We also found CB receptors in cortical and spinal motor neurons. These observations support that targeting this receptor may serve for developing neuroprotective therapies in MNDs.
我们研究了运动神经元病（MND）患者运动皮层中的内源性大麻素系统。
使用MND患者和对照组的尸检样本进行内源性大麻素相关元素的免疫染色和/或蛋白质印迹分析。
我们未在MND患者的运动皮层中发现任何神经元丢失的证据，但胶质细胞标志物Iba-1和GFAP明显升高。我们发现脂肪酸酰胺水解酶（FAAH）和单酰甘油脂肪酶（MAGL）以及CB受体没有变化，这与皮质神经元死亡的缺乏相关。相比之下，蛋白质印迹分析证明运动皮层中CB受体增加，免疫染色也证实了这一点，这与这些患者中胶质细胞增生增加相关。双重标记分析显示，这种升高的CB受体免疫染色位于GFAP标记的星形胶质细胞中。然而，我们也通过与神经元标志物MAP-2的双重免疫荧光证实了皮质神经元中有CB受体标记。在脊髓中也发现了这种情况，使用脊髓运动神经元标志物胆碱乙酰转移酶进行双重标记。患者和对照组均出现这种情况，尽管这些神经元在患者中经历了重要的退化，表现为尼氏染色减少、TDP-43免疫染色减少以及蛋白质印迹法检测的CB受体水平降低。
我们证实MND患者运动皮层中CB受体升高与反应性胶质细胞增生有关。这种现象先于神经元丢失。我们还在皮质和脊髓运动神经元中发现了CB受体。这些观察结果支持靶向该受体可能有助于开发针对运动神经元病的神经保护疗法。
