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在小鼠出生后的早期发育过程中,肠神经系统中优先表达的是DM20,其表达似乎依赖于一个内含子增强子。

in the enteric nervous system is preferentially expressed during early postnatal development in mouse as DM20, whose expression appears reliant on an intronic enhancer.

作者信息

Patyal Pankaj, Fil Daniel, Wight Patricia A

机构信息

Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR, United States.

出版信息

Front Cell Neurosci. 2023 May 24;17:1175614. doi: 10.3389/fncel.2023.1175614. eCollection 2023.

Abstract

Recently, the myelin proteolipid protein gene () was shown to be expressed in the glia of the enteric nervous system (ENS) in mouse. However, beyond this, not much is known about its expression in the intestine. To address this matter, we investigated expression at the mRNA and protein levels in the intestine of mice at different ages (postnatal days 2, 9, 21, and 88). In this study, we show that expression preferentially occurs during early postnatal development, primarily as the DM20 isoform. Western blot analysis indicated that DM20 migrated according to its formula weight when isolated from the intestine. However, mobilities of both PLP and DM20 were faster than expected when procured from the brain. The 6.2hPLP(+)Z/FL transgene, which uses the first half of the human gene to drive expression of a reporter gene, recapitulated the developmental pattern observed with the native gene in the intestine, indicating that it can be used as a proxy for gene expression. As such, the relative levels of β-galactosidase (β-gal) activity emanating from the 6.2hPLP(+)Z/FL transgene suggest that expression is highest in the duodenum, and decreases successively along the segments, toward the colon. Moreover, removal of the wmN1 enhancer region from the transgene (located within intron 1) resulted in a dramatic reduction in both transgene mRNA levels and β-gal activity in the intestine, throughout development, suggesting that this region contains a regulatory element crucial for expression. This is consistent with earlier studies in both the central and peripheral nervous systems, indicating that it may be a common (if not universal) means by which gene expression is governed.

摘要

最近,髓磷脂蛋白脂蛋白基因()被证明在小鼠肠神经系统(ENS)的神经胶质细胞中表达。然而,除此之外,人们对其在肠道中的表达了解不多。为了解决这个问题,我们研究了不同年龄(出生后第2、9、21和88天)小鼠肠道中该基因在mRNA和蛋白质水平的表达情况。在本研究中,我们发现该基因的表达在出生后早期发育阶段优先发生,主要以DM20异构体的形式存在。蛋白质免疫印迹分析表明,从肠道分离出的DM20根据其分子量迁移。然而,从大脑中获取的PLP和DM20的迁移率都比预期的要快。6.2hPLP(+)Z/FL转基因利用人类该基因的前半部分来驱动报告基因的表达,重现了在肠道中观察到的与天然基因相同的发育模式,表明它可以用作该基因表达的替代物。因此,来自6.2hPLP(+)Z/FL转基因的β-半乳糖苷酶(β-gal)活性的相对水平表明,该基因在十二指肠中的表达最高,并沿各节段依次降低,直至结肠。此外,从转基因(位于内含子1内)中去除wmNl增强子区域导致在整个发育过程中肠道中转基因mRNA水平和β-gal活性都显著降低,这表明该区域含有对该基因表达至关重要的调控元件。这与中枢神经系统和外周神经系统早期的研究一致,表明这可能是一种常见(即使不是普遍)的调控该基因表达的方式。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/57ae/10244531/0947abd7d97d/fncel-17-1175614-g0001.jpg

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