Erokhov Pavel A, Lyupina Yulia V, Radchenko Alexandra S, Kolacheva Anna A, Nikishina Yulia O, Sharova Natalia P
Laboratory of Biochemistry of Ontogenesis Processes, N.K. Koltsov Institute of Developmental Biology of Russian Academy of Sciences, Moscow, Russia.
Laboratory of Neural and Neuroendocrine Regulations, N.K. Koltsov Institute of Developmental Biology of Russian Academy of Sciences, Moscow, Russia.
Oncotarget. 2017 Aug 10;8(41):70941-70957. doi: 10.18632/oncotarget.20208. eCollection 2017 Sep 19.
The aim of this work was to detect changes in proteasome pools of brain parts of August rats with monoamine metabolism violations in comparison with that of control Wistar rats. To reveal active proteasome structures, a method of native electrophoresis for the analysis of crude tissue fractions was developed. By means of this method and following Western blotting, the most pronounced changes in reorganization of proteasome structures were detected in proteasome pool of the brain cortex of August rats. Main findings are the enhanced expression of immune proteasome subtypes containing proteolytic subunit LMP2 and activator PA28αβ as well as immune proteasome subtypes containing proteolytic subunit LMP7 and activator PA700 and simultaneously decreased expression of subtypes with subunit LMP2 and activator PA700 in the brain cortex of August rats compared to that of Wistar rats. These results were indirectly confirmed by SDS PAGE method followed by Western blotting, which showed the increased quantities of immune subunits and proteasome activators in the brain cortex of August rats compared to that of Wistar rats. Immune proteasomes were revealed by immunohistochemistry in neurons, but not in glial cells of August and Wistar rat cortex. The detected reorganization of proteasome pools is likely to be important for production of special peptides to provide the steady interaction between neurons and adaptation of central nervous system to conditions caused by monoamine metabolism deviations.
本研究旨在检测与对照Wistar大鼠相比,单胺代谢异常的八月龄大鼠脑区蛋白酶体库的变化。为揭示活性蛋白酶体结构,开发了一种用于分析粗组织组分的非变性电泳方法。通过该方法及后续的蛋白质印迹法,在八月龄大鼠大脑皮层的蛋白酶体库中检测到蛋白酶体结构重组最明显的变化。主要发现是,与Wistar大鼠相比,八月龄大鼠大脑皮层中含有蛋白水解亚基LMP2和激活剂PA28αβ的免疫蛋白酶体亚型以及含有蛋白水解亚基LMP7和激活剂PA700的免疫蛋白酶体亚型表达增强,同时含有亚基LMP2和激活剂PA700的亚型表达降低。SDS-PAGE法及后续蛋白质印迹法间接证实了这些结果,该方法显示与Wistar大鼠相比,八月龄大鼠大脑皮层中免疫亚基和蛋白酶体激活剂的数量增加。通过免疫组织化学在八月龄和Wistar大鼠皮层的神经元而非胶质细胞中发现了免疫蛋白酶体。检测到的蛋白酶体库重组可能对特殊肽的产生很重要,以提供神经元之间的稳定相互作用,并使中枢神经系统适应单胺代谢偏差引起的状况。