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000276854 0247_ $$2doi$$a10.1073/pnas.1611684114
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000276854 0247_ $$2ISSN$$a1091-6490
000276854 037__ $$aDZNE-2025-00367
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000276854 1001_ $$00000-0001-9893-0473$$aMilatz, Susanne$$b0
000276854 245__ $$aMosaic expression of claudins in thick ascending limbs of Henle results in spatial separation of paracellular Na+ and Mg2+ transport.
000276854 260__ $$aWashington, DC$$bNational Acad. of Sciences$$c2017
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000276854 520__ $$aThe thick ascending limb (TAL) of Henle's loop drives paracellular Na+, Ca2+, and Mg2+ reabsorption via the tight junction (TJ). The TJ is composed of claudins that consist of four transmembrane segments, two extracellular segments (ECS1 and -2), and one intracellular loop. Claudins interact within the same (cis) and opposing (trans) plasma membranes. The claudins Cldn10b, -16, and -19 facilitate cation reabsorption in the TAL, and their absence leads to a severe disturbance of renal ion homeostasis. We combined electrophysiological measurements on microperfused mouse TAL segments with subsequent analysis of claudin expression by immunostaining and confocal microscopy. Claudin interaction properties were examined using heterologous expression in the TJ-free cell line HEK 293, live-cell imaging, and Förster/FRET. To reveal determinants of interaction properties, a set of TAL claudin protein chimeras was created and analyzed. Our main findings are that (i) TAL TJs show a mosaic expression pattern of either cldn10b or cldn3/cldn16/cldn19 in a complex; (ii) TJs dominated by cldn10b prefer Na+ over Mg2+, whereas TJs dominated by cldn16 favor Mg2+ over Na+; (iii) cldn10b does not interact with other TAL claudins, whereas cldn3 and cldn16 can interact with cldn19 to form joint strands; and (iv) further claudin segments in addition to ECS2 are crucial for trans interaction. We suggest the existence of at least two spatially distinct types of paracellular channels in TAL: a cldn10b-based channel for monovalent cations such as Na+ and a spatially distinct site for reabsorption of divalent cations such as Ca2+ and Mg2.
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000276854 650_7 $$2Other$$aFRET
000276854 650_7 $$2Other$$aclaudin interaction
000276854 650_7 $$2Other$$amicroperfusion
000276854 650_7 $$2Other$$aparacellular ion transport
000276854 650_7 $$2Other$$atight junction
000276854 650_7 $$2NLM Chemicals$$aClaudins
000276854 650_7 $$09NEZ333N27$$2NLM Chemicals$$aSodium
000276854 650_7 $$0I38ZP9992A$$2NLM Chemicals$$aMagnesium
000276854 650_2 $$2MeSH$$aAnimals
000276854 650_2 $$2MeSH$$aClaudins: genetics
000276854 650_2 $$2MeSH$$aClaudins: metabolism
000276854 650_2 $$2MeSH$$aHEK293 Cells
000276854 650_2 $$2MeSH$$aHumans
000276854 650_2 $$2MeSH$$aLoop of Henle: metabolism
000276854 650_2 $$2MeSH$$aLoop of Henle: physiology
000276854 650_2 $$2MeSH$$aMagnesium: metabolism
000276854 650_2 $$2MeSH$$aMice, Inbred C57BL
000276854 650_2 $$2MeSH$$aMice, Knockout
000276854 650_2 $$2MeSH$$aRats, Sprague-Dawley
000276854 650_2 $$2MeSH$$aSodium: metabolism
000276854 650_2 $$2MeSH$$aTight Junctions: metabolism
000276854 7001_ $$00000-0002-2910-6728$$aHimmerkus, Nina$$b1
000276854 7001_ $$aWulfmeyer, Vera Christine$$b2
000276854 7001_ $$aDrewell, Hoora$$b3
000276854 7001_ $$aMutig, Kerim$$b4
000276854 7001_ $$aHou, Jianghui$$b5
000276854 7001_ $$0P:(DE-2719)9003035$$aBreiderhoff, Tilman$$b6$$udzne
000276854 7001_ $$aMüller, Dominik$$b7
000276854 7001_ $$aFromm, Michael$$b8
000276854 7001_ $$aBleich, Markus$$b9
000276854 7001_ $$aGünzel, Dorothee$$b10
000276854 773__ $$0PERI:(DE-600)1461794-8$$a10.1073/pnas.1611684114$$gVol. 114, no. 2$$n2$$pE219 - E227$$tProceedings of the National Academy of Sciences of the United States of America$$v114$$x0027-8424$$y2017
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