Laboratory of Molecular Neurobiology, Academy of Biology and Biotechnology, Southern Federal University, 194/1 Stachky pr., Rostov-on-Don, 344090, Russia.
Institute of Arid Zones, Southern Scientific Center of Russian Academy of Sciences, 41 Chekhov prosp., Rostov-on-Don, 344006, Russia.
Mol Neurobiol. 2017 Aug;54(6):4172-4188. doi: 10.1007/s12035-016-9964-5. Epub 2016 Jun 21.
After ischemic stroke, cell damage propagates from infarct core to surrounding tissues (penumbra). To reveal proteins involved in neurodegeneration and neuroprotection in penumbra, we studied protein expression changes in 2-mm ring around the core of photothrombotic infarct induced in the rat brain cortex by local laser irradiation after administration of Bengal Rose. The ultrastructural study showed edema and degeneration of neurons, glia, and capillaries. Morphological changes gradually decreased across the penumbra. Using the antibody microarrays, we studied changes in expression of >200 neuronal proteins in penumbra 4 or 24 h after focal photothrombotic infarct. Diverse cellular subsystems were involved in the penumbra tissue response: signal transduction pathways such as protein kinase Bα/GSK-3, protein kinase C and its β1 and β2 isoforms, Wnt/β-catenin (axin1, GSK-3, FRAT1), Notch/NUMB, DYRK1A, TDP43; mitochondria quality control (Pink1, parkin, HtrA2); ubiquitin-mediated proteolysis (ubiquilin-1, UCHL1); axon outgrowth and guidance (NAV-3, CRMP2, PKCβ2); vesicular trafficking (syntaxin-8, TMP21, Munc-18-3, synip, ALS2, VILIP1, syntaxin, synaptophysin, synaptotagmin); biosynthesis of neuromediators (tryptophan hydroxylase, monoamine oxidase B, glutamate decarboxylase, tyrosine hydroxylase, DOPA decarboxylase, dopamine transporter); intercellular interactions (N-cadherin, PMP22); cytoskeleton (neurofilament 68, neurofilament-M, doublecortin); and other proteins (LRP1, prion protein, β-amyloid). These proteins are involved in neurodegeneration or neuroprotection. Such changes were most expressed 4 h after photothrombotic impact. Immunohistochemical and Western blot studies of expression of monoamine oxidase B, UCHL1, DYRK1A, and Munc-18-3 confirmed the proteomic data. These data provide the integral view on the penumbra response to photothrombotic infarct. Some of these proteins can be potential targets for ischemic stroke therapy.
缺血性中风后，细胞损伤从梗塞核心向周围组织（半影区）传播。为了揭示半影区中神经退行性变和神经保护相关的蛋白，我们在给予 Bengal Rose 后，通过局部激光照射诱导大鼠大脑皮质光血栓性梗塞的核心 2 毫米环内，研究了蛋白表达的变化。超微结构研究显示神经元、神经胶质和毛细血管水肿和变性。形态学变化逐渐穿过半影区减少。使用抗体微阵列，我们研究了 4 或 24 小时后局部光血栓性梗塞后半影区中 >200 种神经元蛋白表达的变化。多种细胞亚系统参与了半影组织的反应：信号转导途径，如蛋白激酶 Bα/GSK-3、蛋白激酶 C 及其β1 和β2 同工型、Wnt/β-catenin（轴蛋白 1、GSK-3、FRAT1）、Notch/NUMB、DYRK1A、TDP43；线粒体质量控制（Pink1、parkin、HtrA2）；泛素介导的蛋白水解（泛素蛋白 1、UCHL1）；轴突生长和导向（NAV-3、CRMP2、PKCβ2）；囊泡运输（突触素-8、TMP21、Munc-18-3、synip、ALS2、VILIP1、突触素、突触小体、突触结合蛋白）；神经递质合成（色氨酸羟化酶、单胺氧化酶 B、谷氨酸脱羧酶、酪氨酸羟化酶、多巴胺脱羧酶、多巴胺转运体）；细胞间相互作用（N-钙粘蛋白、PMP22）；细胞骨架（神经丝 68、神经丝-M、双皮质素）；和其他蛋白（LRP1、朊病毒蛋白、β-淀粉样蛋白）。这些蛋白参与神经退行性变或神经保护。这些变化在光血栓性影响后 4 小时表达最为明显。单胺氧化酶 B、UCHL1、DYRK1A 和 Munc-18-3 的免疫组织化学和 Western blot 研究证实了蛋白质组学数据。这些数据提供了对半影区对光血栓性梗塞反应的整体看法。其中一些蛋白可能成为缺血性中风治疗的潜在靶点。
