Li Shenyang, Greuel Brian T, Meng Fanxue, Pereira Glauber B, Pitts Adria, Dobretsova Anna, Wight Patricia A
Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
Gene. 2009 May 1;436(1-2):30-6. doi: 10.1016/j.gene.2009.02.003. Epub 2009 Feb 13.
Although the myelin proteolipid protein gene (Plp1) is highly expressed in the central nervous system encoding the most abundant myelin protein in oligodendrocytes, it is also expressed in other tissues, including testis. Transgenic studies with mice that harbor Plp1-lacZ fusion genes suggest that Leydig cells are the source of Plp1 gene expression in testis. However, virtually nothing is known about Plp1 gene regulation in Leydig cells, which is the focus of this study. The first intron contains both positive and negative regulatory elements that are important in regulating Plp1 gene expression in oligodendrocytes. To test whether these elements are functional in Leydig cells, a battery of Plp1-lacZ fusion genes with partial deletion of Plp1 intron 1 sequence was transfected into the mouse Leydig cell line, TM3. Results presented here suggest that an enhancer, which is very potent in oligodendrocytes, is only nominally active in TM3 cells. The intron also contains several negative regulatory elements that are operative in TM3 cells. Moreover a new exon (exon 1.2) was identified within the first 'intron' resulting in novel splice variants in TM3 cells. Western blot analysis suggests that these splice variants, along with those containing another alternatively spliced exon (exon 1.1) derived from intron 1 sequence, give rise to multiple Plp1 gene products in the mouse testis.
尽管髓磷脂蛋白脂蛋白基因(Plp1)在中枢神经系统中高度表达,该基因编码少突胶质细胞中最丰富的髓磷脂蛋白,但它也在包括睾丸在内的其他组织中表达。对携带Plp1 - lacZ融合基因的小鼠进行的转基因研究表明,睾丸间质细胞是睾丸中Plp1基因表达的来源。然而,关于睾丸间质细胞中Plp1基因的调控,实际上我们几乎一无所知,而这正是本研究的重点。第一个内含子包含正负调控元件,这些元件在少突胶质细胞中对Plp1基因表达的调控起着重要作用。为了测试这些元件在睾丸间质细胞中是否具有功能,将一系列部分缺失Plp1内含子1序列的Plp1 - lacZ融合基因转染到小鼠睾丸间质细胞系TM3中。此处呈现的结果表明,一个在少突胶质细胞中非常有效的增强子在TM3细胞中仅具有微弱的活性。该内含子还包含几个在TM3细胞中起作用的负调控元件。此外,在第一个“内含子”中鉴定出一个新的外显子(外显子1.2),导致TM3细胞中出现新的剪接变体。蛋白质印迹分析表明,这些剪接变体,连同那些包含另一个源自内含子1序列的可变剪接外显子(外显子1.1)的变体,在小鼠睾丸中产生多种Plp1基因产物。