为了研究表达N-甲基-D-天冬氨酸受体(NMDAR)的突触和突触外神经元中24小时表达的长链非编码RNA(lncRNAs),和正常的神经元培养,通过微阵列分析。
将来自胚胎(E18天)Sprague-Dawley大鼠的皮质神经元用于原代神经元培养。阻断NMDAR活化,然后将细胞孵育6小时。提取总RNA,量化,并分析纯度和完整性。双链cDNA合成,其次是分位数归一化,定量聚合酶链反应验证,和数据分析。通过Pearson相关性分析转录因子与lncRNAs之间的相互作用。
在大鼠皮质神经元培养物的突触和突触外NMDAR激活24小时后获得lncRNA谱。总的来说,251个lncRNAs持续上调,335被下调,与正常神经元相比,突触外NMDAR激活后。突触NMDAR激活后,只有9个lncRNAs上调,2个下调。
突触和突触外NMDAR激活后lncRNAs的差异表达表明lncRNAs可能是突触外NMDAR诱导的神经变性的原因。
To study the 24-hour expression of long noncoding RNAs (lncRNAs) in synaptic and extrasynaptic neurons expressing N-methyl-D-aspartate receptor (NMDAR), and normal neuronal cultures, via microarray analysis.
Cortical neurons from embryonic (day E18) Sprague-Dawley rats were used for primary neuronal culture. NMDAR activation was blocked and the cells were then incubated for 6 hours. Total RNA was extracted, quantified, and analyzed for purity and integrity. Double-stranded cDNA was synthesized, followed by quantile normalization, quantitative polymerase chain reaction validation, and data analysis. The interactions between transcription factors and lncRNAs were analyzed by Pearson correlation.
The lncRNA profiles were obtained after synaptic and extrasynaptic NMDAR activation of rat cortical neuron cultures for 24 hours. In total, 251 lncRNAs were consistently upregulated, and 335 were downregulated, after extrasynaptic NMDAR activation compared with normal neurons. After synaptic NMDAR activation, only 9 lncRNAs were upregulated and 2 were downregulated.
Differential expression of lncRNAs after synaptic and extrasynaptic NMDAR activation suggests that lncRNAs may be responsible for extrasynaptic NMDAR-induced neurodegeneration.