Elucidation of the distal convoluted tubule transcriptome identifies new candidate genes involved in renal Mg2+ handling
Elucidation of the distal convoluted tubule transcriptome identifies new candidate genes involved in renal Mg2+ handling
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DOI：
10.1152/ajprenal.00322.2013
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发表时间：
2013-12-01
影响因子：
4.2
通讯作者：
Hoenderop, Joost G. J.
中科院分区：
文献类型：
--
作者：
de Baaij, Jeroen H. F.;Koerkamp, Marian J. Groot;Hoenderop, Joost G. J.
肾脏在维持Mg 2+稳态中起着关键作用。具体而言，远曲小管（DCT）有助于肾Mg 2+处理的微调。近年来，遗传性Mg 2+转运障碍有助于确定DCT Mg 2+稳态的重要参与者。然而，涉及DCT介导的Mg 2+重吸收的几种蛋白质仍有待发现，这个复杂的肾单位片段的完整表达谱可能有助于发现新的Mg 2+相关基因。在这里，我们报告Mg 2+敏感的DCT转录组的表达。为此，将在DCT特异性小清蛋白启动子下表达增强的绿色荧光蛋白的转基因小鼠进行Mg 2+缺乏或Mg 2+富集的饮食。随后，复杂对象参数分析仪和分选仪允许，第一次，增强的绿色荧光蛋白阳性DCT细胞的隔离。通过DNA微阵列分析其RNA提取物，比较高Mg 2+与低Mg 2+，以鉴定Mg 2+调节基因。基于统计学显著性和至少2倍的变化，46个基因显示差异表达。几个已知的亲镁基因，如瞬时受体电位阳离子通道，亚家族M，成员6（Trpm 6），和Parvalbumin，在低饮食Mg 2+上调。此外，还发现了可能参与肾Mg 2+处理的新基因。为了证实所选择的候选基因受膳食Mg 2+可用性的调节，通过RT-PCR分析确定溶质载体家族41，成员3（Slc 41 a3）、蝶呤-4 α-甲醇胺脱氢酶/肝细胞核因子-1 α的二聚化辅因子（Pcbd 1）、TBC 1结构域家族，成员4（Tbc 1d 4）和尿调蛋白（Umod）的表达水平。事实上，所有四个基因在喂食Mg 2+缺乏饮食的小鼠的DCT中显示出显著的上调。通过阐明Mg 2+敏感的DCT转录组，已经鉴定了肾Mg 2+处理的新候选基因。
The kidney plays a key role in the maintenance of Mg2+ homeostasis. Specifically, the distal convoluted tubule (DCT) is instrumental in the fine-tuning of renal Mg2+ handling. In recent years, hereditary Mg2+ transport disorders have helped to identify important players in DCT Mg2+ homeostasis. Nevertheless, several proteins involved in DCT-mediated Mg2+ reabsorption remain to be discovered, and a full expression profile of this complex nephron segment may facilitate the discovery of new Mg2+-related genes. Here, we report Mg2+-sensitive expression of the DCT transcriptome. To this end, transgenic mice expressing enhanced green fluorescent protein under a DCT-specific parvalbumin promoter were subjected to Mg2+-deficient or Mg2+-enriched diets. Subsequently, the Complex Object Parametric Analyzer and Sorter allowed, for the first time, isolation of enhanced green fluorescent protein-positive DCT cells. RNA extracts thereof were analyzed by DNA microarrays comparing high versus low Mg2+ to identify Mg2+ regulatory genes. Based on statistical significance and a fold change of at least 2, 46 genes showed differential expression. Several known magnesiotropic genes, such as transient receptor potential cation channel, subfamily M, member 6 (Trpm6), and Parvalbumin, were upregulated under low dietary Mg2+. Moreover, new genes were identified that are potentially involved in renal Mg2+ handling. To confirm that the selected candidate genes were regulated by dietary Mg2+ availability, the expression levels of solute carrier family 41, member 3 (Slc41a3), pterin-4 alpha-carbinolamine dehydratase/dimerization cofactor of hepatocyte nuclear factor-1 alpha (Pcbd1), TBC1 domain family, member 4 (Tbc1d4), and uromodulin (Umod) were determined by RT-PCR analysis. Indeed, all four genes show significant upregulation in the DCT of mice fed a Mg2+-deficient diet. By elucidating the Mg2+-sensitive DCT transcriptome, new candidate genes in renal Mg2+ handling have been identified.