Pendrin gene ablation alters ENaC subcellular distribution and open probability

Pech, Vladimir; Wall, Susan M.; Nanami, Masayoshi; Bao, Hui‐Fang; Kim, Young‐Hee; Lazo‐Fernandez, Yoskaly; Yue, Qiang; Pham, Truyen D.; Eaton, Douglas C.; Verlander, Jill W.

doi:10.1152/ajprenal.00564.2014

Cited by 35 publications

(66 citation statements)

References 43 publications

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“…Perfusion-fixed mouse kidneys were labeled immunocytochemically (36) or immunohistochemically (37) using the following antibodies: c-Kit (Cell Signaling Technology, Cat. # 3074), V-ATPase A-subunit (36), EPS8 (Santa Cruz, sc-390257), Fosb (Santa Cruz, sc-398595), Foxpl (Santa Cruz, sc-398811), Hoxd8 (Santa Cruz, sc-515357), Scin (Santa Cruz, sc-376136), Tfap2b (Santa Cruz, sc-390119), AE4 (38), pendrin (37), AQP2 (Knepper laboratory, K5007).…”

Section: Methodsmentioning

confidence: 99%

Transcriptomes of major renal collecting-duct cell types in mouse identified by single-cell RNA-Seq

Chen

Lee

Chou

et al. 2017

Preprint

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Prior RNA sequencing (RNA-Seq) studies have identified complete transcriptomes for most renal epithelial cell types. The exceptions are the cell types that make up the renal collecting duct, namely intercalated cells (ICs) and principal cells (PCs), which account for only a small fraction of the kidney mass, but play critical physiological roles in the regulation of blood pressure, extracellular fluid volume and extracellular fluid composition. To enrich these cell types, we used fluorescence-activated cell sorting (FACS) that employed well established lectin cell surface markers for PCs and type B ICs, as well as a newly identified cell surface marker for type A ICs, viz. c-Kit. Single-cell RNA-Seq using the 1C- and PC-enriched populations as input enabled identification of complete transcriptomes of A-ICs, B-ICs and PCs. The data were used to create a freely-accessible online gene-expression database for collecting duct cells. This database allowed identification of genes that are selectively expressed in each cell type including cell-surface receptors, transcription factors, transporters and secreted proteins. The analysis also identified a small fraction of hybrid cells expressing both aquapor¡n-2 and either anion exchanger 1 or pendrin transcripts. In many cases, mRNAs for receptors and their ligands were identified in different cells (e.g. Notch2 chiefly in PCs vs Jag1 chiefly in ICs) suggesting signaling crosstalk among the three cell types. The identified patterns of gene expression among the three types of collecting duct cells provide a foundation for understanding physiological regulation and pathophysiology in the renal collecting duct.SIGNIFICANCE STATEMENTA long-term goal in mammalian biology is to identify the genes expressed in every cell type of the body. In kidney, the expressed genes (“transcriptome”) of all epithelial cell types have already been identified with the exception of the cells that make up the renal collecting duct, responsible for regulation of blood pressure and body fluid composition. Here, a technique called "single-cell RNA-Seq" was used in mouse to identify transcriptomes for the major collecting-duct cell types: type A intercalated cells, type B intercalated cells and principal cells. The information was used to create a publicly-accessible online resource. The data allowed identification of genes that are selectively expressed in each cell type, informative for cell-level understanding of physiology and pathophysiology.

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Section: Methodsmentioning

confidence: 99%

Transcriptomes of major renal collecting-duct cell types in mouse identified by single-cell RNA-Seq

Chen

Lee

Chou

et al. 2017

Preprint

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“…The existence of such cross-talk was expected given that in vitro, variations in extracellular pH or Cldirectly affect ENaC activity (11,12), suggesting that changes in acid/base transporter activity in intercalated cells can impact ENaC-mediated Na + reabsorption. Indeed, studies in WT and pendrin-null mice have demonstrated that both the cell surface expression and the open probability of ENaC are upregulated by pendrin (32,59). Pendrin modulates ENaC abundance and activity by, at least in part, increasing the luminal concentration of HCO 3 and/or pH (57); the underlying molecular mechanisms have yet to be determined.…”

Section: Interactions Between Principal Cells and Type B Cellsmentioning

confidence: 99%

Versatility of NaCl transport mechanisms in the cortical collecting duct

Edwards

Crambert

2017

American Journal of Physiology-Renal Physiology

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The cortical collecting duct (CCD) forms part of the aldosterone-sensitive distal nephron and plays an essential role in maintaining the NaCl balance and acid-base status. The CCD epithelium comprises principal cells as well as different types of intercalated cells. Until recently, transcellular Na transport was thought to be restricted to principal cells, whereas (acid-secreting) type A and (bicarbonate-secreting) type B intercalated cells were associated with the regulation of acid-base homeostasis. This review describes how this traditional view has been upended by several discoveries in the past decade. A series of studies has shown that type B intercalated cells can mediate electroneutral NaCl reabsorption by a mechanism involving Na-dependent and Na-independent Cl/[Formula: see text] exchange, and that is energetically driven by basolateral vacuolar H-ATPase pumps. Other research indicates that type A intercalated cells can mediate NaCl secretion, through a bumetanide-sensitive pathway that is energized by apical H,K-ATPase type 2 pumps operating as Na/K exchangers. We also review recent findings on the contribution of the paracellular route to NaCl transport in the CCD. Last, we describe cross-talk processes, by which one CCD cell type impacts Na/Cl transport in another cell type. The mechanisms that have been identified to date demonstrate clearly the interdependence of NaCl and acid-base transport systems in the CCD. They also highlight the remarkable versatility of this nephron segment.

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“…Ϫ exchange, because this cell type is found primarily in the CNT of the mouse kidney (86) and because mouse CNT is very difficult to perfuse in vitro, CNT transport properties are not well understood. Principal cells mediate the absorption of Na ϩ primarily through the epithelial Na ϩ channel, ENaC (53). However, in the CCD and most probably in the CNT as well, significant Cl Ϫ absorption occurs through transcellular transport across type B and non-A, non-B intercalated cells (64), such as through apical electroneutral Na ϩ -independent, Cl Ϫ /HCO 3 Ϫ exchange (31,63,86,87).…”

Section: Intercalated Cell Subtypes and Their Role In Acid-base Regulmentioning

confidence: 99%

“…In particular, Na ϩ /H ϩ exchanger isoform 3 (NHE3), ␣1-Na-K-ATPase, Na ϩ -K ϩ -2Cl Ϫ cotransporter (NKCC2), ENaC, and NCC abundance was similar in lysates from wild-type and Slc26a4 null mice following a balanced, NaCl-replete diet (32). However, following dietary NaCl restriction or with an aldosterone infusion, pendrin gene ablation reduces ␣-, ␤and ␥-ENaC subunit abundance in the kidney (32,50,53). In particular, pendrin gene ablation blunts the aldosterone-induced increment in 70-kDa ␥-ENaC abundance.…”

Section: Pendrin Modulates the Aldosterone-induced Increment In Enac mentioning

confidence: 99%

“…Since pendrin gene ablation reduces ENaC subunit abundance, we explored ENaC function in wild-type and pendrin null mice. Administration of aldosterone in vivo markedly stimulates Na ϩ absorption in the rat and mouse CCD, which is nearly eliminated with the application of an ENaC inhibitor (53,74). However, the aldosterone-induced increment in Na ϩ absorption observed in the CCD is dependent on pendrin expression.…”

Section: Pendrin Modulates the Aldosterone-induced Increment In Enac mentioning

confidence: 99%

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The role of pendrin in blood pressure regulation

Wall

2016

American Journal of Physiology-Renal Physiology

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Pendrin is a Na(+)-independent Cl(-)/HCO3(-) exchanger found in the apical regions of type B and non-A, non-B intercalated cells within the aldosterone-sensitive region of the nephron, i.e., the distal convoluted tubule (DCT), the connecting tubule (CNT), and the cortical collecting duct (CCD). Type B intercalated cells mediate Cl(-) absorption and HCO3(-) secretion primarily through pendrin-mediated Cl(-)/HCO3(-) exchange. This exchanger is upregulated with angiotensin II administration and in models of metabolic alkalosis, such as following administration of aldosterone or NaHCO3. In the absence of pendrin-mediated HCO3(-) secretion, an enhanced alkalosis is observed following aldosterone or NaHCO3 administration. However, probably of more significance is the role of pendrin in the pressor response to aldosterone. Pendrin mediates Cl(-) absorption and modulates aldosterone-induced Na(+) absorption mediated by the epithelial Na channel (ENaC). Pendrin changes ENaC activity by changing both channel open probability (Po) and surface density (N), at least partly by altering luminal HCO3(-) and ATP concentration. Thus aldosterone and angiotensin II stimulate pendrin expression and function, which stimulates ENaC activity, thereby contributing to the pressor response of these hormones. However, pendrin may modulate blood pressure partly through its extrarenal effects. For example, pendrin is expressed in the adrenal medulla, where it modulates catecholamine release. The increase in catecholamine release observed with pendrin gene ablation likely contributes to the increment in vascular contractile force observed in the pendrin null mouse. This review summarizes the signaling mechanisms that regulate pendrin abundance and function as well as the contribution of pendrin to distal nephron function.

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Pendrin gene ablation alters ENaC subcellular distribution and open probability

Cited by 35 publications

References 43 publications

Transcriptomes of major renal collecting-duct cell types in mouse identified by single-cell RNA-Seq

Transcriptomes of major renal collecting-duct cell types in mouse identified by single-cell RNA-Seq

Versatility of NaCl transport mechanisms in the cortical collecting duct

The role of pendrin in blood pressure regulation

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