PLP1-lacZ transgenic mice reveal that splice variants containing "human-specific" exons are relatively minor in comparison to the archetypal transcript and that an upstream regulatory element bolsters expression during early postnatal brain development.
PLP1-lacZ transgenic mice reveal that splice variants containing "human-specific" exons are relatively minor in comparison to the archetypal transcript and that an upstream regulatory element bolsters expression during early postnatal brain development.
复制标题
DOI：
10.3389/fncel.2022.1087145
复制
发表时间：
2022
影响因子：
5.3
通讯作者：
Wight, Patricia A.
中科院分区：
文献类型：
--
作者：
Patyal, Pankaj;Fil, Daniel;Hamdan, Hamdan;Wight, Patricia A.
关键词：
许多关于控制髓鞘蛋白脂质蛋白基因（PLP 1）的发育表达的机制的已知已经通过使用转基因动物模型获得。在这项研究中，我们分析了相关转基因的表达，这些转基因利用来自人类或小鼠的PLP 1基因组DNA来驱动lacZ报告基因的表达。人PLP 1（hPLP 1）序列跨越外显子2的近端6.2或2.7 kb的5′-侧翼DNA到内部位点，而来自小鼠的那些包含外显子2的近端2.3 kb的5′-侧翼DNA到类似位点。具有hPLP 1序列的转基因部分地根据它们具有的上游序列的量来命名[6.2hPLP（+）Z/FL和2.7hPLP（+）Z]。含有转基因的小鼠序列在此称为mPLP（+）Z，以表示PLP 1 DNA的物种来源。携带6.2hPLP（+）Z/FL转基因的小鼠被用作模型系统，以研究剪接变体的发育表达，所述剪接变体包含来自经典定义为PLP 1内含子1的补充外显子。虽然通过RT-PCR分析在脑中检测到剪接变体的表达，但它们相对于原型（经典）转录物以低得多的水平存在。此外，我们发现携带6.2hPLP（+）Z/FL转基因的小鼠在P2时在整个大脑中表现出广泛的表达，而mPLP（+）Z的表达在这个年龄是非常有限的。因此，我们用2.7hPLP（+）Z转基因产生了新的转基因小鼠品系，其含有与mPLP（+）Z中的hPLP 1序列正向同源的hPLP 1序列。在分析的7个品系中，6个品系在P21与P2相比在脑中显示出更高水平的2.7hPLP（+）Z表达;另一个品系在任一年龄均仅微弱地表达转基因。这种趋势，加上在P2时观察到的6.2hPLP（+）Z/FL的稳健表达，表明对6.2hPLP（+）Z/FL特异的5′侧翼PLP 1 DNA的远端3.5 kb含有对促进脑中出生后早期表达重要的调控元件。
Much of what is known about the mechanisms that control the developmental expression of the myelin proteolipid protein gene (PLP1) has been attained through use of transgenic animal models. In this study, we analyzed expression of related transgenes which utilize PLP1 genomic DNA from either human or mouse to drive expression of a lacZ reporter. Human PLP1 (hPLP1) sequence span either the proximal 6.2 or 2.7 kb of 5′-flanking DNA to an internal site in Exon 2, while those from mouse comprise the proximal 2.3 kb of 5′-flanking DNA to an analogous site in Exon 2. Transgenes with hPLP1 sequence were named, in part, to the amount of upstream sequence they have [6.2hPLP(+)Z/FL and 2.7hPLP(+)Z]. The transgene containing mouse sequence is referred to here as mPLP(+)Z, to denote the species origin of PLP1 DNA. Mice which harbor the 6.2hPLP(+)Z/FL transgene were used as a model system to investigate the developmental expression of splice variants that incorporate supplementary exons from what is classically defined as PLP1 intron 1. While expression of the splice variants were detected in brain through RT-PCR analysis, they are present at much lower levels relative to the archetypal (classic) transcript. Additionally, we show that mice which harbor the 6.2hPLP(+)Z/FL transgene demonstrate wide-ranging expression throughout brain at P2, whereas expression of mPLP(+)Z is quite limited at this age. Therefore, we generated new transgenic mouse lines with the 2.7hPLP(+)Z transgene, which contains hPLP1 sequence orthologous to just that in mPLP(+)Z. Of the seven lines analyzed, six showed higher levels of 2.7hPLP(+)Z expression in brain at P21 compared to P2; the other line expressed the transgene, only weakly, at either age. This trend, coupled with the robust expression observed for 6.2hPLP(+)Z/FL at P2, suggests that the distal 3.5 kb of 5′-flanking PLP1 DNA specific to 6.2hPLP(+)Z/FL contains regulatory element(s) important for promoting early postnatal expression in brain.
登录
查看更多内容
影响因子：
4.4
作者：
Patyal, Pankaj;Kockara, Neriman T.;Wight, Patricia A.
通讯作者：
Wight, Patricia A.
影响因子：
3.3
作者：
Sarret, C.;Combes, P.;Vaurs-Barriere, C.
通讯作者：
Vaurs-Barriere, C.
DOI：
10.1083/jcb.123.2.443
发表时间：
1993-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Wight PA;Duchala CS;Readhead C;Macklin WB
通讯作者：
Macklin WB
影响因子：
2.9
作者：
YOUNG, DC;KINGSLEY, SD;DUTKO, FJ
通讯作者：
DUTKO, FJ
DOI：
10.1073/pnas.84.16.5665
发表时间：
1987-08-01
影响因子：
11.1
作者：
NAVE, KA;LAI, C;MILNER, RJ
通讯作者：
MILNER, RJ