Demonstration of the existence of mRNAs encoding N1/cif and N2/cit sodium/nucleoside cotransporters in rat brain
Demonstration of the existence of mRNAs encoding N1/cif and N2/cit sodium/nucleoside cotransporters in rat brain
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DOI：
10.1016/s0169-328x(96)00244-6
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发表时间：
1996-12-01
期刊：
影响因子：
--
通讯作者：
Parkinson, FE
中科院分区：
文献类型：
--
作者：
Anderson, CM;Xiong, W;Parkinson, FE
核苷转运可能参与细胞外腺苷水平的调节，腺苷是中枢神经系统中的抑制性神经调节剂。先前的报告为大鼠大脑中 Na+ 依赖性核苷转运提供了功能证据。我们从大鼠大脑的各个区域分离了总 RNA，并使用逆转录酶 PCR (RT-PCR) 测试了两个最近克隆的 Na+/核苷协同转运蛋白的 mRNA 的存在。通过 RT-PCR 扩增 309 bp DNA 产物，在每个大脑区域的样本中检测到嘧啶选择性 Na+/核苷协同转运蛋白 mRNA (rCNT1) 的信使 RNA。 Southern印迹和序列分析证实该产物源自rCNT1 mRNA。通过扩增 235 bp DNA 产物（其序列与已发表的序列相同），在整个大脑中检测到嘌呤选择性 Na+/核苷协同转运蛋白 mRNA（rCNT2，也称为 SPNT）。这些实验证明大鼠大脑中存在 rCNT1 和 rCNT2 mRNA。
Nucleoside transport may be involved in the regulation of extracellular levels of adenosine, an inhibitory neuromodulator in the central nervous system. Previous reports have provided functional evidence for Na+-dependent nucleoside transport in rat brain. We isolated total RNA from various regions of rat brain and tested for the presence of mRNA for two recently cloned Na+/nucleoside cotransporters using reverse transcriptase PCR (RT-PCR). Messenger RNA for a pyrimidine-selective Na+/nucleoside cotransporter mRNA (rCNT1) was detected in samples from each brain region tested by RT-PCR amplification of a 309-bp DNA product. Southern blot and sequence analysis confirmed that this product was derived from rCNT1 mRNA. A purine-selective Na+/nucleoside cotransporter mRNA (rCNT2, also termed SPNT) was detected throughout brain by amplifying a 235-bp DNA product, the sequence of which was identical to that published. These experiments demonstrate the presence of both rCNT1 and rCNT2 mRNA in rat brain.