Ferrari Federica, Gorini Antonella, Villa Roberto Federico
Laboratory of Pharmacology and Molecular Medicine of Central Nervous System, Department of Biology and Biotechnology, University of Pavia, Via Ferrata, 9, 27100, Pavia, Italy.
Neurochem Res. 2015 Jan;40(1):172-85. doi: 10.1007/s11064-014-1482-0. Epub 2014 Nov 30.
Functional proteomics was used to characterize age-related changes in energy metabolism of different neuronal pathways within the cerebellar cortex of Wistar rats aged 2, 6, 12, 18, and 24 months. The "large" synaptosomes, derived from the glutamatergic mossy fibre endings which make synaptic contact with the granule cells of the granular layer, and the "small" synaptosomes, derived from the pre-synaptic terminals of granule cells making synaptic contact with the dendrites of Purkinje cells, were isolated by a combined differential/gradient centrifugation technique. Because most brain disorders are associated with bioenergetic changes, the maximum rate (Vmax) of selected enzymes of glycolysis, Krebs' cycle, glutamate and amino acids metabolism, and acetylcholine catabolism were evaluated. The results show that "large" and "small" synaptosomes possess specific and independent metabolic features. This study represents a reliable model to study in vivo (1) the physiopathological molecular mechanisms of some brain diseases dependent on energy metabolism, (2) the responsiveness to noxious stimuli, and (3) the effects of drugs, discriminating their action sites at subcellular level on specific neuronal pathways.
功能蛋白质组学被用于表征2、6、12、18和24月龄Wistar大鼠小脑皮质内不同神经通路能量代谢的年龄相关变化。通过联合差速/梯度离心技术分离出“大”突触体，其来源于与颗粒层颗粒细胞形成突触联系的谷氨酸能苔藓纤维末梢；以及“小”突触体，其来源于与浦肯野细胞树突形成突触联系的颗粒细胞突触前终末。由于大多数脑部疾病都与生物能量变化有关，因此评估了糖酵解、三羧酸循环、谷氨酸和氨基酸代谢以及乙酰胆碱分解代谢中所选酶的最大反应速率（Vmax）。结果表明，“大”和“小”突触体具有特定且独立的代谢特征。本研究是一个可靠的模型，可用于在体内研究：（1）某些依赖能量代谢的脑部疾病的生理病理分子机制；（2）对有害刺激的反应性；（3）药物的作用，在亚细胞水平区分它们在特定神经通路上的作用位点。
