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黄颡鱼(Pelteobagrus fulvidraco)中IIb型钠依赖性磷酸盐共转运体(Slc34a2)亚型的特征及其在低磷条件下维生素D调节的表达

Characterization of the isoforms of type IIb sodium-dependent phosphate cotransporter (Slc34a2) in yellow catfish, Pelteobagrus fulvidraco, and their vitamin D-regulated expression under low-phosphate conditions.

作者信息

Chen Pei, Huang Yanqing, Bayir Abdulkadir, Wang Chunfang

机构信息

The College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.

Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, People's Republic of China.

出版信息

Fish Physiol Biochem. 2017 Feb;43(1):229-244. doi: 10.1007/s10695-016-0282-7. Epub 2016 Sep 12.

Abstract

In this study, two isoforms slc34a2 genes (type IIb sodium-dependent phosphate cotransporter), slc34a2a2 and slc34a2b, were cloned from intestine and kidney of yellow catfish (Pelteobagrus fulvidraco), with rapid amplification of cDNA ends. The structure differences and the regulation effects of dietary VD under low phosphorus were compared among three isoforms of slc34a2 in yellow catfish. The predicted Slc34a2a2 and Slc34a2b proteins match 65 % and 53.8 % sequence identity, with Slc34a2a1, respectively. The membrane-spanning domains were different among these three isoforms. Intestinal Slc34a2a1 and Slc34a2a2 proteins had eight and eleven transmembrane domains, while renal Slc34a2b protein had nine. The tissue distribution study showed that same as slc34a2a1, slc34a2a2 mRNA was mainly distributed in intestine and slc34a2b mRNA in kidney. The effect of vitamin D (VD) level on slc34a2 subfamily expression under low-phosphate conditions, induced by the addition of 0 (VD0), 324 (VD1), 1243 (VD2), 3621 (VD3), 8040 (VD4), or 22700 (VD5) IU VD/kg feed, was assessed by qPCR. The dose-responsive expression of intestinal slc34a2a2 and high expression of intestinal slc34a2a2 in VD5 together with peak expression of kidney slc34a2b in VD3 coincided with the accumulation of body phosphate content. These data suggested that appropriate level of dietary VD up-regulated slc34a2a1, slc34a2a2, and slc34a2b mRNA levels, which increased phosphate retention. In conclusion, the current study provided another possible approach to improve dietary phosphate utilization by adding appropriate level of VD to a low-phosphate diet to regulate intestinal and renal slc34a2 gene expression and thus minimize the excretion of phosphorus in yellow catfish.

摘要

在本研究中,通过cDNA末端快速扩增技术,从黄颡鱼(Pelteobagrus fulvidraco)的肠道和肾脏中克隆出两种IIb型钠依赖性磷酸盐共转运体(slc34a2)基因亚型,即slc34a2a2和slc34a2b。比较了黄颡鱼slc34a2三种亚型的结构差异以及低磷条件下日粮维生素D(VD)的调节作用。预测的Slc34a2a2和Slc34a2b蛋白与Slc34a2a1的序列同一性分别为65%和53.8%。这三种亚型的跨膜结构域不同。肠道中的Slc34a2a1和Slc34a2a2蛋白分别有8个和11个跨膜结构域,而肾脏中的Slc34a2b蛋白有9个。组织分布研究表明,与slc34a2a1一样,slc34a2a2 mRNA主要分布在肠道,slc34a2b mRNA主要分布在肾脏。通过qPCR评估了添加0(VD0)、324(VD1)、1243(VD2)、3621(VD3)、8040(VD4)或22700(VD5)IU VD/kg饲料诱导的低磷条件下VD水平对slc3亚家族表达的影响。肠道slc34a2a2的剂量反应性表达以及VD5组中肠道slc34a2a2的高表达,与VD3组中肾脏slc34a2b的峰值表达,均与鱼体磷含量的积累一致。这些数据表明,日粮中适当水平的VD上调了slc34a2a1、slc34a2a2和slc34a2b的mRNA水平,从而增加了磷的潴留。总之,本研究提供了另一种可能的方法,即通过在低磷日粮中添加适当水平的VD来调节肠道和肾脏slc34a2基因表达,从而减少黄颡鱼磷的排泄,提高日粮磷的利用率。

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