Walpole Caragh, McGrane Alison, Al-Mousawi Hashemeya, Winter Desmond, Baird Alan, Stewart Gavin
School of Biology & Environmental Science, University College Dublin, Dublin, Ireland.
Institute for Clinical Outcomes Research and Education, St. Vincent's University Hospital, Dublin, Ireland.
Physiol Rep. 2018 Aug;6(15):e13826. doi: 10.14814/phy2.13826.
The symbiotic relationship between humans and their intestinal microbiome is supported by urea nitrogen salvaging. Previous studies have shown that colonic UT-B urea transporters play a significant role in this important physiological process. This current study investigated UT-A and UT-B urea transporter expression along the human gastrointestinal tract. Initial end-point PCR experiments determined that UT-A RNA was predominantly expressed in the small intestine, while UT-B RNA was expressed in stomach, small intestine, and colon. Using western blotting experiments, a strong 40-60 kDa UT-B signal was found to be abundant in both ileum and colon. Importantly, this signal was deglycosylated by PNGaseF enzyme treatment to a core protein of 30 kDa in both tissues. Further immunolocalization studies revealed UT-B transporter proteins were present at the apical membrane of the villi in the ileum, but predominantly at the basolateral membrane of the colonic surface epithelial cells. Finally, a blind scoring immunolocalization study suggested that there was no significant difference in UT-B abundance throughout the colon (NS, ANOVA, N = 5-21). In conclusion, this current study suggested UT-B to be the main human intestinal urea transporter. Intriguingly, these data suggested that the same UT-B isoform was present in all intestinal epithelial cells, but that the precise cellular location varied.
尿素氮的回收利用支持了人类与其肠道微生物群之间的共生关系。先前的研究表明,结肠UT-B尿素转运体在这一重要生理过程中发挥着重要作用。本研究调查了UT-A和UT-B尿素转运体在人类胃肠道中的表达情况。最初的终点PCR实验确定,UT-A RNA主要在小肠中表达,而UT-B RNA在胃、小肠和结肠中表达。通过蛋白质印迹实验发现,在回肠和结肠中均大量存在一条强的40-60 kDa的UT-B信号。重要的是,用PNGaseF酶处理后,该信号在两种组织中均被去糖基化至30 kDa的核心蛋白。进一步的免疫定位研究表明,UT-B转运蛋白存在于回肠绒毛的顶端膜,但主要存在于结肠表面上皮细胞的基底外侧膜。最后,一项盲法评分免疫定位研究表明,整个结肠中UT-B的丰度没有显著差异(无显著性差异,方差分析,N = 5-21)。总之,本研究表明UT-B是人类肠道主要的尿素转运体。有趣的是,这些数据表明,相同的UT-B同工型存在于所有肠道上皮细胞中,但精确的细胞定位有所不同。