Identification and cataloging of genes induced by long-lasting long-term potentiation in awake rats
Identification and cataloging of genes induced by long-lasting long-term potentiation in awake rats
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DOI：
10.1046/j.1471-4159.2000.0742239.x
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发表时间：
2000-06-01
影响因子：
4.7
通讯作者：
Inokuchi, K
中科院分区：
文献类型：
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作者：
Matsuo, R;Murayama, A;Inokuchi, K
长时程增强（LTP）的维持需要从头基因表达。在这里，我们报告的直接分离，使用PCR-差异显示，基因的表达水平发生变化后，诱导长期的LTP在自由活动的清醒大鼠海马。使用480个引物组合的差异显示显示17个cDNA条带，显示了可重复的表达水平的变化。这些cDNA代表了至少10个不同的基因（称为RM 1 -10），所有这些基因在LTP诱导后75 min均显示上调，并在24 h内恢复到基础表达水平。这些基因中的三个仅从表达的序列标签（RM 1 -3）已知，两个是其通过LTP的上调尚未被描述的已知基因（GADD 153/ CHOP和ler 5），并且五个是其通过LTP的上调已经被报道的已知基因（MAPK磷酸酶、NGFI-A/zif 268、vesl-1 S/homer-1a、Ag 2和krox-20）。我们采用北方印迹法和半定量RT-PCR技术，研究了前两类基因在NMDA受体依赖性、组织特异性和发育调控方面的表达谱。所有这五个基因的上调是NMDA受体依赖性的，并与LTP的持久性相关，这表明这些基因可能在长期的LTP维持中发挥功能性作用。
Maintenance of long-term potentiation (LTP) requires de novo gene expression. Here we report the direct isolation, using PCR-differential display, of genes whose expression level was altered after induction of long-lasting LTP in the hippocampus of freely moving awake rats. Differential display using 480 primer combinations revealed 17 cDNA bands that showed a reproducible change in expression level. These cDNAs represented at least 10 different genes (termed RM1-10), all of which showed up-regulation at 75 min after LTP induction and a return to basal expression levels within 24 h. Three of these genes were known only from expressed sequence tags (RM1-3), two were known genes whose up-regulation by LTP has not been described (GADD 153/ CHOP and ler5), and five were known genes whose upregulation by LTP has already been reported (MAPK phosphatase, NGFI-A/zif268, vesl-1S/homer-1a, Ag2, and krox-20). We characterized the expression profiles of genes in the two former categories with respect to NMDA receptor dependency, tissue specificity, and developmental regulation using northern blotting and semiquantitative RT-PCR. The up-regulation of all five of these genes was NMDA receptor-dependent and correlated with the persistence of LTP, suggesting that these genes may play functional roles in prolonged LTP maintenance.