Expression and function of the lipocalin-2 (24p3/NGAL) receptor in rodent and human intestinal epithelia.

The lipocalin 2//NGAL/24p3 receptor (NGAL-R/24p3-R) is expressed in rodent distal nephron where it mediates protein endocytosis. The mechanisms of apical endocytosis and transcytosis of proteins and peptides in the intestine are poorly understood. In the present study, the expression and localizatio...

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Autores principales:	Christian Langelueddecke, Eleni Roussa, Robert A Fenton, Frank Thévenod
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/84fbaa96374645859a233a5fedd2ef87
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spelling	oai:doaj.org-article:84fbaa96374645859a233a5fedd2ef872021-11-18T09:01:08ZExpression and function of the lipocalin-2 (24p3/NGAL) receptor in rodent and human intestinal epithelia.1932-620310.1371/journal.pone.0071586https://doaj.org/article/84fbaa96374645859a233a5fedd2ef872013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23940770/?tool=EBIhttps://doaj.org/toc/1932-6203The lipocalin 2//NGAL/24p3 receptor (NGAL-R/24p3-R) is expressed in rodent distal nephron where it mediates protein endocytosis. The mechanisms of apical endocytosis and transcytosis of proteins and peptides in the intestine are poorly understood. In the present study, the expression and localization of rodent 24p3-R (r24p3-R) and human NGAL-R (hNGAL-R) was investigated in intestinal segments by immunofluorescence and confocal laser scanning microscopy, immunohistochemistry and immunoblotting. r24p3-R/hNGAL-R was also studied in human Caco-2 BBE cells and CHO cells transiently transfected with r24p3-R by immunofluorescence microscopy, RT-PCR and immunoblotting of plasma membrane enriched vesicles (PM). To assay function, endocytosis/transcytosis of putative ligands phytochelatin (PC₃), metallothionein (MT) and transferrin (Tf) was assayed by measuring internalization of fluorescence-labelled ligands in Caco-2 BBE cells grown on plastic or as monolayers on Transwell inserts. The binding affinity of Alexa 488-PC₃ to colon-like Caco-2 BBE PM was quantified by microscale thermophoresis (MST). r24p3-R/hNGAL-R expression was detected apically in all intestinal segments but showed the highest expression in ileum and colon. Colon-like, but not duodenum-like, Caco-2 BBE cells expressed hNGAL-R on their surface. Colon-like Caco-2 BBE cells or r24p3-R transfected CHO cells internalized fluorescence-labelled PC₃ or MT with half-maximal saturation at submicromolar concentrations. Uptake of PC₃ and MT (0.7 µM) by Caco-2 BBE cells was partially blocked by hNGAL (500 pM) and an EC₅₀ of 18.6 ± 12.2 nM was determined for binding of Alexa 488-PC₃ to PM vesicles by MST. Transwell experiments showed rapid (0.5-2 h) apical uptake and basolateral delivery of fluorescent PC₃/MT/Tf (0.7 µM). Apical uptake of ligands was significantly blocked by 500 pM hNGAL. hNGAL-R dependent uptake was more prominent with MT but transcytosis efficiency was reduced compared to PC₃ and Tf. Hence, r24p3-R/hNGAL-R may represent a high-affinity multi-ligand receptor for apical internalization and transcytosis of intact proteins/peptides by the lower intestine.Christian LangelueddeckeEleni RoussaRobert A FentonFrank ThévenodPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 8, p e71586 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Christian Langelueddecke Eleni Roussa Robert A Fenton Frank Thévenod Expression and function of the lipocalin-2 (24p3/NGAL) receptor in rodent and human intestinal epithelia.
description	The lipocalin 2//NGAL/24p3 receptor (NGAL-R/24p3-R) is expressed in rodent distal nephron where it mediates protein endocytosis. The mechanisms of apical endocytosis and transcytosis of proteins and peptides in the intestine are poorly understood. In the present study, the expression and localization of rodent 24p3-R (r24p3-R) and human NGAL-R (hNGAL-R) was investigated in intestinal segments by immunofluorescence and confocal laser scanning microscopy, immunohistochemistry and immunoblotting. r24p3-R/hNGAL-R was also studied in human Caco-2 BBE cells and CHO cells transiently transfected with r24p3-R by immunofluorescence microscopy, RT-PCR and immunoblotting of plasma membrane enriched vesicles (PM). To assay function, endocytosis/transcytosis of putative ligands phytochelatin (PC₃), metallothionein (MT) and transferrin (Tf) was assayed by measuring internalization of fluorescence-labelled ligands in Caco-2 BBE cells grown on plastic or as monolayers on Transwell inserts. The binding affinity of Alexa 488-PC₃ to colon-like Caco-2 BBE PM was quantified by microscale thermophoresis (MST). r24p3-R/hNGAL-R expression was detected apically in all intestinal segments but showed the highest expression in ileum and colon. Colon-like, but not duodenum-like, Caco-2 BBE cells expressed hNGAL-R on their surface. Colon-like Caco-2 BBE cells or r24p3-R transfected CHO cells internalized fluorescence-labelled PC₃ or MT with half-maximal saturation at submicromolar concentrations. Uptake of PC₃ and MT (0.7 µM) by Caco-2 BBE cells was partially blocked by hNGAL (500 pM) and an EC₅₀ of 18.6 ± 12.2 nM was determined for binding of Alexa 488-PC₃ to PM vesicles by MST. Transwell experiments showed rapid (0.5-2 h) apical uptake and basolateral delivery of fluorescent PC₃/MT/Tf (0.7 µM). Apical uptake of ligands was significantly blocked by 500 pM hNGAL. hNGAL-R dependent uptake was more prominent with MT but transcytosis efficiency was reduced compared to PC₃ and Tf. Hence, r24p3-R/hNGAL-R may represent a high-affinity multi-ligand receptor for apical internalization and transcytosis of intact proteins/peptides by the lower intestine.
format	article
author	Christian Langelueddecke Eleni Roussa Robert A Fenton Frank Thévenod
author_facet	Christian Langelueddecke Eleni Roussa Robert A Fenton Frank Thévenod
author_sort	Christian Langelueddecke
title	Expression and function of the lipocalin-2 (24p3/NGAL) receptor in rodent and human intestinal epithelia.
title_short	Expression and function of the lipocalin-2 (24p3/NGAL) receptor in rodent and human intestinal epithelia.
title_full	Expression and function of the lipocalin-2 (24p3/NGAL) receptor in rodent and human intestinal epithelia.
title_fullStr	Expression and function of the lipocalin-2 (24p3/NGAL) receptor in rodent and human intestinal epithelia.
title_full_unstemmed	Expression and function of the lipocalin-2 (24p3/NGAL) receptor in rodent and human intestinal epithelia.
title_sort	expression and function of the lipocalin-2 (24p3/ngal) receptor in rodent and human intestinal epithelia.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/84fbaa96374645859a233a5fedd2ef87
work_keys_str_mv	AT christianlangelueddecke expressionandfunctionofthelipocalin224p3ngalreceptorinrodentandhumanintestinalepithelia AT eleniroussa expressionandfunctionofthelipocalin224p3ngalreceptorinrodentandhumanintestinalepithelia AT robertafenton expressionandfunctionofthelipocalin224p3ngalreceptorinrodentandhumanintestinalepithelia AT frankthevenod expressionandfunctionofthelipocalin224p3ngalreceptorinrodentandhumanintestinalepithelia
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Expression and function of the lipocalin-2 (24p3/NGAL) receptor in rodent and human intestinal epithelia.

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