Ben-Ari Shani, Toiber Debra, Sas Aldema S, Soreq Hermona, Ben-Shaul Yoram
Department of Biological Chemistry and Israel Center for Neuronal Computation, The Hebrew University of Jerusalem, Jerusalem, Israel.
J Neurochem. 2006 Apr;97 Suppl 1:24-34. doi: 10.1111/j.1471-4159.2006.03725.x.
Alternative splicing configurations and acetylcholinesterase (AChE) gene expression are both modified in neurons under stress. However, it is unclear if these phenomena are functionally interrelated. Using a home-made spotted microarray focused on splicing-associated transcripts, we tested the effects of excess 3' splice variants of human AChE on splicing-related gene expression in semi-differentiated neuronal P19 cells. Of the tested transcripts, 17.3% and 20.2% showed modified expression levels (log2 of the ratio<-0.3 or>0.3) in transfected P19 cells overexpressing the stress-inducible AChE-R variant or the synaptic AChE-S protein, respectively. Multiple transcripts encoding serine-arginine rich (SR) and SR-related splicing regulators were suppressed in cells expressing either of these variants, whereas the gene groups including splicing-related helicases and transcripts involved in apoptosis displayed variant-specific changes. Our findings are compatible with the assumption that both neuronal overexpression and alternative splicing of pre-AChE mRNA may be causally involved in initiating global changes in neuronal alternative splicing, causing subsequent modifications in the expression patterns of numerous target genes.
在应激状态下，神经元中的可变剪接构型和乙酰胆碱酯酶（AChE）基因表达均会发生改变。然而，尚不清楚这些现象在功能上是否相互关联。我们使用一种自制的聚焦于剪接相关转录本的点阵微阵列，测试了人AChE过量的3'剪接变体对半分化神经元P19细胞中剪接相关基因表达的影响。在测试的转录本中，分别有17.3%和20.2%在过表达应激诱导型AChE-R变体或突触型AChE-S蛋白的转染P19细胞中显示出表达水平的改变（比值的log2<-0.3或>0.3）。在表达这两种变体之一的细胞中，多个编码富含丝氨酸-精氨酸（SR）和SR相关剪接调节因子的转录本受到抑制，而包括剪接相关解旋酶和参与凋亡的转录本在内的基因组则表现出变体特异性变化。我们的研究结果与以下假设相符：AChE前体mRNA的神经元过表达和可变剪接都可能因果性地参与引发神经元可变剪接的全局变化，进而导致众多靶基因表达模式的后续改变。
