Department of Biological Sciences, Key Laboratory of Molecular Biophysics of Ministry of Education, Huazhong University of Science & Technology School of Life Science & Technology, Wuhan, Hubei Province 430074, China.
Brain Res. 2011 May 16;1390:33-40. doi: 10.1016/j.brainres.2011.03.046. Epub 2011 Mar 23.
The SLC4A10 gene, which is highly expressed in the mammalian brain, contains two known alternative splicing units, inserts A and B, and is theoretically capable of producing four NBCn2 splice variants: NBCn2-A, -B, -C, and -D. By immunoprecipitation and western blotting, a previous study showed the putative NBCn2-D to be expressed predominantly in the subcortex (SCX) and medulla (MD) of mouse brain. However, no evidence has been provided, in any species, for the existence of a full-length transcript encoding NBCn2-D. In the present study, we report for the first time the cloning of the full-length cDNAs encoding NBCn2-D from mouse SCX and MD. Based on the frequency of bacterial colonies obtained after PCR, we conclude that in SCX, the NBCn2-A transcript is dominant, whereas in MD, NBCn2-B is dominant. NBCn2-D is the least abundant transcript in each of these two brain regions. An analysis based upon the present PCR data as well as the previous immunoprecipitation/western-blot data suggests the following prevalence of NBCn2 variants in total mouse brain: NBCn2-A (83%), NBCn2-B (10%), NBCn2-C (5%), and NBCn2-D (2%). We also estimate the prevalence of each variant in each of the five brain regions (i.e., cerebral cortex, SCX, cerebellum, hippocampus, and MD). We hypothesize that the expression of different NBCn2 splice variants is characteristic of specific tissue/cells.
SLC4A10 基因在哺乳动物大脑中高度表达,包含两个已知的选择性剪接单元,插入 A 和 B,理论上能够产生四种 NBCn2 剪接变体:NBCn2-A、-B、-C 和 -D。通过免疫沉淀和 Western blot,之前的一项研究表明,假定的 NBCn2-D 主要在小鼠大脑的皮质下区 (SCX) 和延髓 (MD) 中表达。然而,在任何物种中,都没有提供存在编码 NBCn2-D 的全长转录本的证据。在本研究中,我们首次报道了从小鼠 SCX 和 MD 克隆全长编码 NBCn2-D 的 cDNA。基于 PCR 后获得的细菌菌落频率,我们得出结论,在 SCX 中,NBCn2-A 转录本占优势,而在 MD 中,NBCn2-B 占优势。NBCn2-D 在这两个脑区中都是最不丰富的转录本。基于当前 PCR 数据以及之前的免疫沉淀/Western blot 数据的分析表明,NBCn2 变体在整个小鼠脑中的流行情况如下:NBCn2-A(83%)、NBCn2-B(10%)、NBCn2-C(5%)和 NBCn2-D(2%)。我们还估计了每个变体在五个脑区(即大脑皮层、SCX、小脑、海马体和 MD)中的流行情况。我们假设不同 NBCn2 剪接变体的表达是特定组织/细胞的特征。