Evaluation and Comparison of Vitamin D Responsive Gene Expression in Ovine, Canine and Equine Kidney.
Evaluation and Comparison of Vitamin D Responsive Gene Expression in Ovine, Canine and Equine Kidney.
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DOI：
10.1371/journal.pone.0162598
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Thompson KG
中科院分区：
文献类型：
--
作者：
Azarpeykan S;Dittmer KE;Marshall JC;Perera KC;Gee EK;Acke E;Thompson KG
本研究的目的是利用实时定量聚合酶链式反应(RT-qPCR)技术测定绵羊、犬和马肾脏中维生素D应答转录物和钙转运转录物(TRPV5、TRPV6、calD9k、calD28k、PMCA、NCX1、CYP27B1、CYP24A1和VDR)的相对丰度及其相互关系，并对这三个物种进行比较。肾组织样本是从10匹马、10只羊和5只狗死后采集的。针对每个基因设计了引物。对每个样本提取总RNA，合成cDNA，进行RT-qPCR。对RT-qPCR数据进行归一化处理并进行统计学比较。TRPV6、calD9k/calD28k和PMCA是绵羊、狗和马肾脏主动跨上皮钙转运的主要途径。结果表明，所有研究的基因都在所研究物种的肾脏组织中表达，尽管转录本的表达水平和相互之间的相关性在物种之间存在差异。在马肾中，所有维生素D反应转录本和钙转运转录本都与VDR高度相关，而在绵羊和狗中则不相关。与绵羊和狗相比，马和狗的肾脏表达模式和相关性不同。考虑到马的尿钙浓度较高，而血清1，25(OH)2D浓度较低，因此可以预期，在马的体内，细胞色素P27B1的表达将低于细胞色素P24A1，但情况似乎并非如此。这一发现表明，尽管血清维生素D浓度较低，但维生素D在马的钙代谢中仍然发挥着重要作用，特别是考虑到这些动物中VDR和维生素D反应转录本之间的强烈相关性。
The aim of this study was to determine the relative abundance and relationship of vitamin D responsive and calcium transporting transcripts (TRPV5, TRPV6, calD9k, calD28k, PMCA, NCX1, CYP27B1, CYP24A1, and VDR) in ovine, canine and, equine kidney using quantitative real-time PCR (RT-qPCR), and then perform a comparison between the three species. Renal tissue samples were harvested post-mortem from 10 horses, 10 sheep, and five dogs. Primers were designed for each gene. For each sample total RNA was extracted, cDNA synthesised, and RT-qPCR was performed. RT-qPCR data were normalised and statistical comparison was performed. Due to their consistent correlation with each other in each species, TRPV6, calD9k/calD28k, and PMCA appeared to be the main pathways involved in active transepithelial calcium transport in the kidney of sheep, dogs and horses. The results indicate that all of the studied genes were expressed in the renal tissue of studied species, although the expression levels and correlation of transcripts with each other were different from species to species. All vitamin D responsive and calcium transporting transcripts were highly correlated with VDR in equine kidney, but not in sheep and dogs. The CYP27B1 and CYP24A1 mRNAs showed a different renal expression pattern and correlation in horses compared with sheep and dogs. Given the high urinary calcium concentration and low serum 1,25(OH)2D concentration in horses, it could be expected that CYP27B1 expression would be lower than CYP24A1 in the horse, and this did not appear to be the case. The findings suggest that despite low serum vitamin D concentrations, vitamin D still plays a significant role in calcium metabolism in horses, especially given the strong correlations between VDR and vitamin D responsive transcripts in these animals.
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