Antisense oligonucleotide suppression of Na+/Ca2+ exchanger activity in primary neurons from rat brain
Antisense oligonucleotide suppression of Na+/Ca2+ exchanger activity in primary neurons from rat brain
复制标题
DOI：
10.1016/s0304-3940(00)01524-x
复制
发表时间：
2000-11-10
影响因子：
2.5
通讯作者：
Michaelis, ML
中科院分区：
文献类型：
--
作者：
Ranciat-McComb, NS;Bland, KS;Michaelis, ML
基于Na+/Ca 2+交换器（NCX）的三种亚型中的保守序列的反义（AS）寡脱氧核苷酸被用于降低这种Ca 2+转运体在原代神经元培养物中的表达。两种AS寡核苷酸应用在12-24小时内使NCX活性降低了约40%，并且正义（S）或错义（MS）寡核苷酸都不改变NCX活性。通过免疫印迹和酶联免疫吸附试验（ELISA）证实NCX表达减少。静息[Ca 2 +]（i）水平在AS处理的神经元中高20%，并且在N-甲基-D-天冬氨酸（NMDA）激活Ca 2+内流后显示出缓慢恢复至基线水平。这些结果表明，NCX起着重要的作用，在维持神经元Ca 2+稳态和恢复基线Ca 2+水平后去极化。(C)2000爱思唯尔科学爱尔兰有限公司保留所有权利。
An antisense (AS) oligodeoxynucleotide based on a conserved sequence in the three isoforms of the Na+/Ca2+ exchanger (NCX) was used to decrease expression of this Ca2+ transporter in primary neuronal cultures. Two AS oligo applications decreased NCX activity by similar to 40% within 12-24 h, and neither sense (S) or missense (MS) oligos altered NCX activity. The reduced NCX expression was confirmed by immunoblots and enzyme-linked immunosorbent assays (ELISAs). Resting [Ca2+](i) levels were 20% higher in AS-treated neurons and showed a slower return to baseline levels following activation of Ca2+ influx by N-methyl-D-aspartate (NMDA). These results suggest that NCX plays a significant role in maintaining neuronal Ca2+ homeostasis and in restoring baseline Ca2+ levels following depolarization. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.