Keywords

Rat Hippocampal Neuron, Oxygen And Glucose Deprivation, Drp1 Protein, Cytc, Caspase 3 Rat Hippocampal Neuron, Oxygen And Glucose Deprivation, Drp1 Protein, Cytochrome C (Cytc), Caspase 3

Abstract

Abstract: Objective To observe the changes in mitochondrial morphology and apoptosis-related proteins in rat hippocampal neurons after being treated with oxygen and glucose deprivation (OGD) for different times. Methods Primary cultured rat hippocampal neurons were randomly divided into 5 groups: blank control group (control group) and OGD-treated 30min group, 45min group, 60min group and 90min group. The mitochondrial morphology was observed using laser confocal microscopy; JC-1 Detect changes in mitochondrial membrane potential; Western blot detects intracytoplasmic phosphorylated mitochondrial power-related protein (P-Drp1 protein), cytochrome C (CytC) and cysteine-containing aspartate hydrolyzable protein 3 (caspase 3) protein expression. Results As the OGD treatment time prolongs, ¢Ù the expression of P-Drp1 protein decreases and the mitochondrial lysis rate gradually increases (P<0.05); ¢Ú the mitochondrial membrane potential decreases more seriously; ¢Û compared with the control group, CytC and caspase 3 in the 60min group and 90min group The protein expression was significantly increased (P<0.05). Conclusion As OGD treatment prolongs, mitochondrial fragmentation increases, membrane potential decreases, membrane permeability increases, and apoptosis occurs at 60 minutes after OGD treatment. ABSTRACT: Objective To evaluate the progressive effect of oxygen and glucose deprivation on fission level of mitochondria, transformation of fission-related protein and ultimately apoptosis-related biogenesis in rat hippocampal neurons. Methods Primary cultured neurons were divided into 5 groups randomly: normal control group (OGD 0min, 0min), OGD 30min group (30min), OGD 45min group (45min), OGD 60min group (60min), and OGD 90min group (90min). The neurons were exposed to oxygen and glucose deprivation (OGD) for 0min , 30min, 45min, 60min and 90min. Mitochondria were stained by Mitochondria tracker Deep Red and shot by laser confocal microscopy, and mitochondrial membrane potential was stained by JC-1 and examined by fluorescence microscopy. Assays of P-Drp1, CytC and caspase 3 were quantified by Western blot. Results ¢Ù With prolonged OGD treatment, P-Drp1 protein expression in neurons was obviously decreased and mitochondrial fission rate gradually increased (P<0.05). ¢Ú Mitochondrial transmembrane potential decreased more greatly. ¢Û Cytochrome C (CytC) and caspase 3 proteins had significantly high expression in 60min and 90min groups as compared with control group (P<0.05). Conclusion As OGD treatment prolongs, mitochondrial fission increases, mitochondrial transmembrane potential decreases, and membrane permeability increases, which occur with apoptosis at OGD 60 minutes time point of OGD treatment   For further details regarding this article and its research, please do not hesitate to contact our team for assistance. Original research done by Li Xuehan, Liu Xueliang, Wang Ping, Lu Zhiyu, Wu Yong, Chen Xiu  

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