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Investigation of facilitative urea transporters in the human gastrointestinal tract

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 trans...

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Autores principales: Walpole, Caragh, McGrane, Alison, Al‐mousawi, Hashemeya, Winter, Desmond, Baird, Alan, Stewart, Gavin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6087735/
https://www.ncbi.nlm.nih.gov/pubmed/30101448
http://dx.doi.org/10.14814/phy2.13826
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author Walpole, Caragh
McGrane, Alison
Al‐mousawi, Hashemeya
Winter, Desmond
Baird, Alan
Stewart, Gavin
author_facet Walpole, Caragh
McGrane, Alison
Al‐mousawi, Hashemeya
Winter, Desmond
Baird, Alan
Stewart, Gavin
author_sort Walpole, Caragh
collection PubMed
description 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.
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spelling pubmed-60877352018-08-17 Investigation of facilitative urea transporters in the human gastrointestinal tract Walpole, Caragh McGrane, Alison Al‐mousawi, Hashemeya Winter, Desmond Baird, Alan Stewart, Gavin Physiol Rep Original Research 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. John Wiley and Sons Inc. 2018-08-12 /pmc/articles/PMC6087735/ /pubmed/30101448 http://dx.doi.org/10.14814/phy2.13826 Text en © 2018 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Research
Walpole, Caragh
McGrane, Alison
Al‐mousawi, Hashemeya
Winter, Desmond
Baird, Alan
Stewart, Gavin
Investigation of facilitative urea transporters in the human gastrointestinal tract
title Investigation of facilitative urea transporters in the human gastrointestinal tract
title_full Investigation of facilitative urea transporters in the human gastrointestinal tract
title_fullStr Investigation of facilitative urea transporters in the human gastrointestinal tract
title_full_unstemmed Investigation of facilitative urea transporters in the human gastrointestinal tract
title_short Investigation of facilitative urea transporters in the human gastrointestinal tract
title_sort investigation of facilitative urea transporters in the human gastrointestinal tract
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6087735/
https://www.ncbi.nlm.nih.gov/pubmed/30101448
http://dx.doi.org/10.14814/phy2.13826
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