AP4M1 is abnormally expressed in oxygen-glucose deprived hippocampal neurons
AP4M1 is abnormally expressed in oxygen-glucose deprived hippocampal neurons
复制标题
AP4M1 在缺氧-葡萄糖的海马神经元中异常表达
DOI：
10.1016/j.neulet.2014.01.034
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发表时间：
2014
影响因子：
2.5
通讯作者：
Sheng G. Y.
中科院分区：
文献类型：
--
作者：
Zhang J.;Cheng X. Y.;Sheng G. Y.
AP 4 M1突变与常染色体隐性遗传性脑瘫综合征有关。但其致病机制尚不明确。本研究的目的是研究损伤神经元中AP 4 M1表达是否以及如何变化。原代培养的海马神经元用于本实验。它们经受氧-葡萄糖剥夺（OGD），导致细胞凋亡，模拟脑缺血。神经元特异性烯醇化酶（NSE）免疫荧光标记证实神经元纯度大于90%。实时荧光定量PCR和蛋白质印迹法检测基因表达。用MAP 2或Tau-1标记AP 4 M1，观察其分布。我们发现，在OGD组和假手术组之间，手术后即刻的AP 4 M1蛋白水平相似。再灌注12 h后表达下调，24 h时表达更明显。实时PCR显示类似的结果，除了在OGD后立即能够检测到mRNA的下调。免疫荧光标记显示，正常神经元中AP 4 M1主要分布于树突，而OGD后AP 4 M1重新分布于轴突。总之，在缺氧缺糖海马神经元中，AP 4 M1不仅在mRNA和蛋白水平上下调，而且从树突向轴突重新分布
AP4M1 mutations have been suggested to be associated with autosomal recessive cerebral palsy syndrome. But the pathogenic mechanism remains uncertain. The purpose of this study is to investigate whether and how AP4M1 expression is changed in injured neurons. Primary cultured hippocampal neurons were prepared for this experiment. They were subjected to oxygen–glucose deprivation (OGD) leading to apoptosis, mimicking brain ischemia. Neuron-specific enolase (NSE) was labeled immunofluorescently to confirm that the purity of neuron was higher than 90%. Real-time PCR and western blotting were performed to measure the gene expression. AP4M1 was labeled with MAP2 or Tau-1 to observe the distribution. We found that the AP4M1 protein levels immediately after the procedure were similar between the OGD group and the sham group. However, down-regulation was observed 12 h after the reperfusion, and became more notable at 24 h. The real-time PCR showed similar results, except that the down-regulation of mRNA was able to be detected immediately after the OGD. Immunofluorescent labeling revealed AP4M1 distributed in the dendrites of normal neurons, but it redistributed to the axons after the OGD procedure. In conclusion, AP4M1 is not only down-regulated at both the mRNA and protein levels, but also redistributed from dendrites to axons in oxygen–glucose deprived hippocampal neurons