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1.
Heliyon ; 9(11): e22280, 2023 Nov.
Article in English | MEDLINE | ID: mdl-38034706

ABSTRACT

In the renal collecting ducts, chloride reabsorption occurs through both transcellular and paracellular pathways. Recent literature highlights a functional interplay between both pathways. We recently showed that in polarized inner medullary collecting duct cells, expression of the basolateral kidney anion exchanger 1 (kAE1) results in a decreased transepithelial electrical resistance (TEER), in a claudin-4 dependent pathway. Claudin-4 is a paracellular sodium blocker and chloride pore. Here, we show that kAE1 expression in mouse inner medullary collecting duct cells triggers WNK4, SPAK and claudin-4 phosphorylation. Expression of a functionally dead kAE1 E681Q mutant has no effect on phosphorylation of these proteins. Expression of a catalytically inactive WNK4 D321A or chloride-insensitive WNK4 L319F mutant abolishes kAE1 effect on TEER, supporting a contribution of WNK4 to the process. We propose that variations of the cytosolic pH and chloride concentration upon kAE1 expression alter WNK4 kinase activity and tight junction properties.

2.
Biochim Biophys Acta Biomembr ; 1862(6): 183238, 2020 06 01.
Article in English | MEDLINE | ID: mdl-32119864

ABSTRACT

Acid-secreting intercalated cells of the collecting duct express the chloride/bicarbonate kidney anion exchanger 1 (kAE1) as well as SLC26A7, two proteins that colocalize in the basolateral membrane. The latter protein has been reported to function either as a chloride/bicarbonate exchanger or a chloride channel. Both kAE1 and SLC26A7 are detected in the renal medulla, an environment hyper-osmotic to plasma. Individuals with mutations in the SLC4A1 gene encoding kAE1 and mice lacking Slc26a7 develop distal renal tubular acidosis (dRTA). Here, we aimed to (i) confirm that SLC26A7 can function as chloride/bicarbonate exchanger in Madin-Darby canine kidney (MDCK) cells, and (ii) examine the behavior of SLC26A7 relative to kAE1 wild type or carrying the dRTA mutation R901X in iso- or hyper-osmotic conditions mimicking the renal medulla. Although we found that SLC26A7 abundance increases in hyper-osmotic growth medium, it is reduced in low pH growth conditions mimicking acidosis when expressed at high levels in MDCK cells. In these cells, SLC26A7 exchange activity was independent from extracellular osmolarity. When SLC26A7 protein was co-expressed with kAE1 WT or the R901X dRTA mutant, the cellular chloride/bicarbonate exchange rate was not additive compared to when proteins are expressed individually, possibly reflecting a decreased overall protein expression. Furthermore, the cellular chloride/bicarbonate exchange rate was osmolarity-independent. Together, these results show that (i) in MDCK cells, SLC26A7 is a chloride/bicarbonate exchanger whose abundance is up-regulated by high osmolarity growth medium and (ii) acidic extracellular pH decreases the abundance of SLC26A7 protein.


Subject(s)
Chloride-Bicarbonate Antiporters/analysis , Hydrogen-Ion Concentration , Kidney/cytology , Osmolar Concentration , Animals , Antiporters/analysis , Cell Culture Techniques/methods , Culture Media/chemistry , Dogs , Epithelial Cells/chemistry , Gene Expression Regulation , Madin Darby Canine Kidney Cells , Sulfate Transporters/analysis
3.
Sci Rep ; 9(1): 3099, 2019 02 28.
Article in English | MEDLINE | ID: mdl-30816203

ABSTRACT

In the renal collecting duct, intercalated cells regulate acid-base balance by effluxing protons through the v-H+-ATPase, and bicarbonate via apical pendrin or the basolateral kidney anion exchanger 1 (kAE1). Additionally, collecting duct cells play an essential role in transepithelial absorption of sodium and chloride. Expression of kAE1 in polarized MDCK I cells was previously shown to decrease trans-epithelial electrical resistance (TEER), suggesting a novel role for kAE1 in paracellular permeability. In our study, we not only confirmed that inducible expression of kAE1 in mIMCD3 cells decreased TEER but we also observed (i) increased epithelial absolute permeability to both sodium and chloride, and (ii) that this effect was dependent on kAE1 activity. Further, kAE1 regulated tight junction properties through the tight junction protein claudin-4, a protein with which it physically interacts and colocalizes. These findings unveil a novel interaction between the junctional protein claudin-4 and the kidney anion exchanger, which may be relevant to ion and/or pH homeostasis.


Subject(s)
Anion Exchange Protein 1, Erythrocyte/physiology , Claudin-4/metabolism , Kidney Tubules, Collecting/cytology , Tight Junctions/metabolism , Animals , Biological Transport , Cell Line , Cell Membrane Permeability , Chlorides/metabolism , Electric Impedance , Kidney/metabolism , Mice , Sodium/metabolism
4.
Biochem Cell Biol ; 97(3): 234-242, 2019 06.
Article in English | MEDLINE | ID: mdl-30208280

ABSTRACT

In the kidney, the collecting duct (CD) is composed of at least four cell types: principal, type-A intercalated cells (IC), type-B IC, and non-A and non-B IC. Although this heterogeneous composition has been recognized since the end of the nineteenth century, the physiological role of the various cell types in the CD continues to be deciphered as of today. Principal and ICs are essential in ion-water balance and acid-base homeostasis, respectively. However, recent research has revealed a striking interplay and overlap between the specific functions of these cell types. This review summarizes the recent findings on CD cells and their role in multiple pathophysiologies.


Subject(s)
Kidney Tubules/metabolism , Kidney/metabolism , Animals , Humans
5.
J Am Soc Nephrol ; 28(5): 1507-1520, 2017 May.
Article in English | MEDLINE | ID: mdl-27932475

ABSTRACT

Distal nephron acid secretion is mediated by highly specialized type A intercalated cells (A-ICs), which contain vacuolar H+-ATPase (V-type ATPase)-rich vesicles that fuse with the apical plasma membrane on demand. Intracellular bicarbonate generated by luminal H+ secretion is removed by the basolateral anion-exchanger AE1. Chronically reduced renal acid excretion in distal renal tubular acidosis (dRTA) may lead to nephrocalcinosis and renal failure. Studies in MDCK monolayers led to the proposal of a dominant-negative trafficking mechanism to explain AE1-associated dominant dRTA. To test this hypothesis in vivo, we generated an Ae1 R607H knockin mouse, which corresponds to the most common dominant dRTA mutation in human AE1, R589H. Compared with wild-type mice, heterozygous and homozygous R607H knockin mice displayed incomplete dRTA characterized by compensatory upregulation of the Na+/HCO3- cotransporter NBCn1. Red blood cell Ae1-mediated anion-exchange activity and surface polypeptide expression did not change. Mutant mice expressed far less Ae1 in A-ICs, but basolateral targeting of the mutant protein was preserved. Notably, mutant mice also exhibited reduced expression of V-type ATPase and compromised targeting of this proton pump to the plasma membrane upon acid challenge. Accumulation of p62- and ubiquitin-positive material in A-ICs of knockin mice suggested a defect in the degradative pathway, which may explain the observed loss of A-ICs. R607H knockin did not affect type B intercalated cells. We propose that reduced basolateral anion-exchange activity in A-ICs inhibits trafficking and regulation of V-type ATPase, compromising luminal H+ secretion and possibly lysosomal acidification.


Subject(s)
Acidosis, Renal Tubular/enzymology , Anion Exchange Protein 1, Erythrocyte/physiology , Kidney Tubules, Collecting/cytology , Kidney Tubules, Collecting/enzymology , Vacuolar Proton-Translocating ATPases/physiology , Animals , Anion Exchange Protein 1, Erythrocyte/genetics , Male , Mice , Models, Biological
6.
Am J Physiol Cell Physiol ; 310(9): C764-72, 2016 05 01.
Article in English | MEDLINE | ID: mdl-26984737

ABSTRACT

Mutations in the SLC4A1 gene coding for kidney anion exchanger 1 (kAE1) cause distal renal tubular acidosis (dRTA). We investigated the fate of the most common truncated dominant dRTA mutant kAE1 R901X. In renal epithelial cells, we found that kAE1 R901X is less abundant than kAE1 wild-type (WT) at the plasma membrane. Although kAE1 WT and kAE1 R901X have similar half-lives, the decreased abundance of kAE1 R901X at the surface is due to an increased endocytosis rate and a decreased recycling rate of endocytosed proteins. We propose that, in polarized renal epithelial cells, the apically mistargeted kAE1 R901X mutant is endocytosed faster than kAE1 WT and its recycling to the basolateral membrane is delayed. This resets the equilibrium, such that kAE1 R901X resides predominantly in an endomembrane compartment, thereby likely participating in development of dRTA disease.


Subject(s)
Anion Exchange Protein 1, Erythrocyte/metabolism , Epithelial Cells/metabolism , Kidney/metabolism , Animals , Anion Exchange Protein 1, Erythrocyte/genetics , Cell Line , Cell Membrane/metabolism , HEK293 Cells , Humans , Kidney/cytology , Mutation/genetics , Structure-Activity Relationship , Swine
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