TAAR1-EAAT2通路抑制乳鼠星形胶质细胞的谷氨酸摄取能力

发布时间：2018-05-07 01:00

本文选题：多巴胺 + 谷氨酸　；参考：《细胞与分子免疫学杂志》2017年07期

【摘要】：目的研究多巴胺(DA)对原代星形胶质细胞谷氨酸(Glu)摄取能力的影响,以及DA通过痕量胺相关受体1-兴奋性氨基酸转运体2(TAAR1-EAAT2)信号通路对星形胶质细胞Glu摄取能力的影响。方法采用Amplex Red谷氨酸测定试剂盒测定经过DA处理的原代星形胶质细胞对Glu摄取含量的变化,反转录PCR检测TAAR1、EAAT2 mRNA水平,Western blot法检测TAAR1、EAAT2蛋白水平;采用TAAR1小干扰RNA(siRNA)和TAAR1质粒转染DA处理后的原代星形胶质细胞,Western blot法检测EAAT2表达水平并采用Amplex Red谷氨酸测定试剂盒测定培养上清液Glu含量。结果 DA处理的原代星形胶质细胞EAAT2水平降低,TAAR1水平增加,培养上清液Glu含量上升。DA处理经TAAR1 siRNA转染后的原代星形胶质细胞,上调EAAT2水平,培养上清液中Glu含量减少;DA处理经TAAR1质粒转染后的原代星形胶质细胞,则EAAT2降低,培养上清液中Glu的含量增加。结论 DA通过影响星形胶质细胞TAAR1-EAAT2信号通路,减弱细胞Glu摄取能力,引起细胞外Glu蓄积,从而损伤星形胶质细胞的功能。
[Abstract]:Objective to study the effect of dopamine (DA) on the uptake of glutamate (Glu1) in primary astrocytes and the effect of DA on the Glu uptake of astrocytes through the signal pathway of trace amine-associated receptor 1 excitatory amino acid transporter 2TAAR1-EAAT2. Methods Amplex Red assay kit was used to detect the changes of Glu uptake by DA treated primary astrocytes, and reverse transcription PCR was used to detect the level of TAAR1 / EAAT2 mRNA and the protein level of TAAR1 / EAAT2 was detected by Western blot. The expression of EAAT2 was detected by TAAR1 small interference RNA-siRNAs and TAAR1 plasmid transfection into DA treated primary astrocytes by Western blot, and the Glu content of culture supernatant was measured by Amplex Red assay kit. Results the EAAT2 level of primary astrocytes treated with DA decreased the level of tact AR1 increased, the Glu content of culture supernatant increased. DA treated primary astrocytes transfected with TAAR1 siRNA increased the level of EAAT2. The content of Glu in culture supernatant was decreased and the content of Glu in supernatant was increased when EAAT2 was decreased when DA was treated with TAAR1 plasmid transfected astrocytes. Conclusion DA can decrease the ability of Glu uptake and induce the accumulation of extracellular Glu through affecting the TAAR1-EAAT2 signaling pathway of astrocytes, thus impairing the function of astrocytes.
【作者单位】：温州医科大学附属第一医院浙江省神经老化与疾病研究重点实验室;温州医科大学附属第一医院外科实验室;温州医科大学附属第一医院神经外科;
【基金】：国家自然科学基金(81671042,81300308)
【分类号】：R741

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