The wmN1 Enhancer Region of the Mouse Myelin Proteolipid Protein Gene (mPlp1) is Indispensable for Expression of an mPlp1-lacZ Transgene in Both the CNS and PNS
The wmN1 Enhancer Region of the Mouse Myelin Proteolipid Protein Gene (mPlp1) is Indispensable for Expression of an mPlp1-lacZ Transgene in Both the CNS and PNS
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DOI：
10.1007/s11064-019-02919-w
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发表时间：
2019-11-28
影响因子：
4.4
通讯作者：
Wight, Patricia A.
中科院分区：
文献类型：
--
作者：
Patyal, Pankaj;Kockara, Neriman T.;Wight, Patricia A.
髓磷脂蛋白脂蛋白基因（PLP1）编码中枢神经系统髓磷脂中最丰富的蛋白。PLP1基因的表达必须严格调控，人类x连锁白质营养不良是由PLP1拷贝数变化引起的，包括剂量升高和缺失。最近，我们使用Cre-lox方法发现，人类PLP1 （hPLP1）内含子1中的wmN1区域是促进小鼠高水平hPLP1- lacz转基因所必需的。目前的研究测试了从含有小鼠Plp1 (mPlp1) DNA的相关转基因中丢失wmN1区域是否会产生类似的结果。此外，我们还研究了mPlp1内含子1中另一个区域（ASE）丢失的影响。先前的研究表明，通过转染分析，ASE是促进mPlp1-lacZ高水平表达所必需的，但在小鼠中从原生基因中去除ASE则没有作用。这是由于mPlp1中另一个未包含在mPlp1- lacz结构中的调控元件的补偿，还是由于方法上的差异，目前尚不清楚。产生了两个携带mPLP(+)Z/FL的转基因小鼠系。亲本转基因利用mPlp1序列（近2.3 kb的5MODIFIER字母prime侧DNA到第2外显子的前37 bp）驱动lacZ报告磁带的表达。本研究证实mPLP(+)Z/FL在大脑少突胶质细胞、少突胶质前体细胞、嗅鞘细胞和神经元以及坐骨神经雪旺细胞中均有表达。亲本转基因中wmN1区域的缺失会抑制表达，而ASE的移除则没有影响。
The myelin proteolipid protein gene (PLP1) encodes the most abundant protein in CNS myelin. Expression of the gene must be strictly regulated, as evidenced by human X-linked leukodystrophies resulting from variations in PLP1 copy number, including elevated dosages as well as deletions. Recently, we showed that the wmN1 region in human PLP1 (hPLP1) intron 1 is required to promote high levels of an hPLP1-lacZ transgene in mice, using a Cre-lox approach. The current study tests whether loss of the wmN1 region from a related transgene containing mouse Plp1 (mPlp1) DNA produces similar results. In addition, we investigated the effects of loss of another region (ASE) in mPlp1 intron 1. Previous studies have shown that the ASE is required to promote high levels of mPlp1-lacZ expression by transfection analysis, but had no effect when removed from the native gene in mouse. Whether this is due to compensation by another regulatory element in mPlp1 that was not included in the mPlp1-lacZ constructs, or to differences in methodology, is unclear. Two transgenic mouse lines were generated that harbor mPLP(+)Z/FL. The parental transgene utilizes mPlp1 sequences (proximal 2.3 kb of 5MODIFIER LETTER PRIME-flanking DNA to the first 37 bp of exon 2) to drive expression of a lacZ reporter cassette. Here we demonstrate that mPLP(+)Z/FL is expressed in oligodendrocytes, oligodendrocyte precursor cells, olfactory ensheathing cells and neurons in brain, and Schwann cells in sciatic nerve. Loss of the wmN1 region from the parental transgene abolished expression, whereas removal of the ASE had no effect.