Beilharz E J, Zhukovsky E, Lanahan A A, Worley P F, Nikolich K, Goodman L J
Lynx Therapeutics, Hayward, California 94545, USA.
J Neurosci. 1998 Dec 1;18(23):9780-9. doi: 10.1523/JNEUROSCI.18-23-09780.1998.
Synaptic activity induces a rapid transcriptional response that is essential for the establishment of long-term neuronal plasticity. Using a differential cloning technique, we have identified a gene induced by seizure activity in the brain as RB3. RB3 is a recently cloned gene belonging to the stathmin family of phosphoproteins. Like SCG10, RB3 is brain-specific, although in situ hybridization results show that the expression of RB3 is more ubiquitous than is that of SCG10. Using genomic DNA sequencing, we show that the 27 amino acid sequence unique to the RB3" transcript is encoded by an alternatively spliced exon, exon 2'. Using a peptide antibody raised against exon 2' to detect RB3" and an anti-Flag antibody to detect an epitope-tagged version of RB3, we show that both proteins are localized to the Golgi apparatus of transfected COS7 cells. Of particular interest, RB3 mRNA, but not SCG10 mRNA, is rapidly induced in the dentate gyrus granule layer of the hippocampus after electrically induced seizure activity as well as stimuli leading to long-term potentiation (LTP). In addition, RB3 mRNA is induced in pheochromocytoma (PC12) cells treated with 250 ng/ml NGF. These results suggest that RB3 may play a role in activity-induced neuronal plasticity and neuronal differentiation.
突触活动会引发快速的转录反应,这对于长期神经元可塑性的建立至关重要。利用差异克隆技术,我们在大脑中鉴定出一个由癫痫活动诱导的基因,即RB3。RB3是最近克隆的一个基因,属于磷蛋白的stathmin家族。与SCG10一样,RB3具有脑特异性,不过原位杂交结果显示,RB3的表达比SCG10更为广泛。通过基因组DNA测序,我们发现RB3“转录本特有的27个氨基酸序列由一个可变剪接外显子——外显子2'编码。使用针对外显子2'产生的肽抗体来检测RB3”,并用抗Flag抗体检测RB3的表位标记版本,我们发现这两种蛋白质都定位于转染的COS7细胞的高尔基体。特别有趣的是,在电诱导癫痫活动以及导致长时程增强(LTP)的刺激后,海马齿状回颗粒层中RB3 mRNA会迅速被诱导,而SCG10 mRNA则不会。此外,用250 ng/ml NGF处理的嗜铬细胞瘤(PC12)细胞中也会诱导RB3 mRNA。这些结果表明,RB3可能在活动诱导的神经元可塑性和神经元分化中发挥作用。