Indraswari Fransisca, Wong Peter T H, Yap Elgin, Ng Y K, Dheen S Thameem
Department of Anatomy, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore.
Neurochem Int. 2009 Sep;55(4):235-42. doi: 10.1016/j.neuint.2009.03.005. Epub 2009 Mar 18.
Ischemia activates the synthesis of potentially damaging and protective proteins in the central nervous system. Dihydropyrimidinase-like 2 (Dpysl2), a protein involved in neuronal differentiation and axonal guidance, and alpha-spectrin 2 (Spna2), a protein involved in maintaining neuronal membrane integrity, were found altered in various nervous system diseases. Modifications of Dpysl2 and Spna2 proteins have been reported in focal ischemic stroke, but their significance is not yet established. Therefore, this study was aimed to investigate the temporal expression of Dpysl2 and Spna2 genes in normal and stroke rat brain and to characterize stroke brains for cell areas, apoptosis, and microglia cells. The middle cerebral artery of rat brain was occluded and the brain tissue was sectioned for in situ hybridization of Dpysl2 and Spna2 genes, TUNEL, and OX-42 immunofluorescence staining. Dpysl2 and Spna2 mRNA expression was quantified by real-time RT-PCR. Characterization of stroke brain for apoptosis and microglia cells showed apoptotic cells and activated microglia, mainly in the infarct core of ipsilateral cortex and striatum of stroke brain. Significant upregulation of Dpysl2 and Spna2 mRNA expression in the penumbra region after stroke was observed predominantly in injured swollen cells in the cortex and striatum. Upregulation of Dpysl2 and Spna2 expression in hypertrophic cells in the penumbra regions of cortex and striatum of stroke brain indicates an early neuronal defense mechanism involving active neuronal repair, regeneration and development, as these genes are known to be involved in neurite outgrowth and plasticity.
缺血会激活中枢神经系统中潜在的损伤性和保护性蛋白质的合成。二氢嘧啶酶样2(Dpysl2)是一种参与神经元分化和轴突导向的蛋白质,α-血影蛋白2(Spna2)是一种参与维持神经元膜完整性的蛋白质,在各种神经系统疾病中均发现其发生了改变。在局灶性缺血性卒中中已报道了Dpysl2和Spna2蛋白的修饰,但它们的意义尚未明确。因此,本研究旨在探讨Dpysl2和Spna2基因在正常和卒中大鼠脑中的时间表达,并对卒中脑的细胞区域、细胞凋亡和小胶质细胞进行特征描述。闭塞大鼠脑的大脑中动脉,并将脑组织切片用于Dpysl2和Spna2基因的原位杂交、TUNEL法以及OX-42免疫荧光染色。通过实时逆转录聚合酶链反应(RT-PCR)对Dpysl2和Spna2 mRNA表达进行定量分析。对卒中脑的细胞凋亡和小胶质细胞的特征描述显示,凋亡细胞和活化的小胶质细胞主要存在于卒中脑同侧皮质和纹状体的梗死核心区域。卒中后在半暗带区域观察到Dpysl2和Spna2 mRNA表达显著上调,主要见于皮质和纹状体中受损肿胀的细胞。卒中脑皮质和纹状体半暗带区域肥大细胞中Dpysl2和Spna2表达上调,表明存在一种早期神经元防御机制,涉及活跃的神经元修复、再生和发育,因为已知这些基因参与神经突生长和可塑性。