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Dietary Na+ depletion up-regulates NKCC1 expression and enhances electrogenic Cl− secretion in rat proximal colon

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Abstract

The corticosteroid hormone, aldosterone, markedly enhances K+ secretion throughout the colon, a mechanism critical to its role in maintaining overall K+ balance. Previous studies demonstrated that basolateral NKCC1 was up-regulated by aldosterone in the distal colon specifically to support K+ secretion—which is distinct from the more well-established role of NKCC1 in supporting luminal Cl− secretion. However, considerable segmental variability exists between proximal and distal colonic ion transport processes, especially concerning their regulation by aldosterone. Furthermore, delineating such region-specific effects has important implications for the management of various gastrointestinal pathologies. Experiments were therefore designed to determine whether aldosterone similarly up-regulates NKCC1 in the proximal colon to support K+ secretion. Using dietary Na+ depletion as a model of secondary hyperaldosteronism in rats, we found that proximal colon NKCC1 expression was indeed enhanced in Na+-depleted (i.e., hyperaldosteronemic) rats. Surprisingly, electrogenic K+ secretion was not detectable by short-circuit current (ISC) measurements in response to either basolateral bumetanide (NKCC1 inhibitor) or luminal Ba2+ (non-selective K+ channel blocker), despite enhanced K+ secretion in Na+-depleted rats, as measured by 86Rb+ fluxes. Expression of BK and IK channels was also found to be unaltered by dietary Na+ depletion. However, bumetanide-sensitive basal and agonist-stimulated Cl− secretion (ISC) were significantly enhanced by Na+ depletion, as was CFTR Cl− channel expression. These data suggest that NKCC1-dependent secretory pathways are differentially regulated by aldosterone in proximal and distal colon. Development of therapeutic strategies in treating pathologies related to aberrant colonic K+/Cl− transport—such as pseudo-obstruction or ulcerative colitis—may benefit from these findings.

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Abbreviations

ENaC:

Epithelial Na+ channel

NKCC1:

Na+/K+/2Cl− co-transporter 1

BK:

Large conductance K+ channel

IK:

Intermediate conductance K+ channel

CFTR:

Cystic fibrosis transmembrane conductance regulator

EDTA:

5 Ethylenediaminetetraacetic acid

DTT:

Dithiothreitol

PMSF:

Phenylmethylsulphonyl fluoride

CCH:

Carbachol

FSK:

Forskolin

ISC :

Short-circuit current

G.I:

Gastrointestinal

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Acknowledgements

We acknowledge the technical assistance rendered by Avinash Elangovan.

Funding

This study was supported by the National Institute of Health NIDDK grants DK104791 and DK112085 grants to VMR.

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Author notes

  1. Andrew J. Nickerson

    Present address: University of Pittsburgh, S929 Scaife Hall, 3550 Terrace Street, Pittsburgh, PA, USA

Authors and Affiliations

  1. Departments of Physiology, Pharmacology and Neuroscience, West Virginia University School of Medicine, Morgantown, WV, USA

    Andrew J. Nickerson

  2. Departments of Biochemistry and Molecular Medicine, West Virginia University School of Medicine, 1 Medical Center Drive, Morgantown, WV, 26506, USA

    Andrew J. Nickerson & Vazhaikkurichi M. Rajendran

  3. Department of Medicine, West Virginia University School of Medicine, Morgantown, WV, USA

    Vazhaikkurichi M. Rajendran

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Contributions

AJN conceived the idea, performed experiments, wrote manuscript. VMR conceived the idea and got funding. Both AJN and VMR discussed and edited the manuscript.

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Correspondence to Vazhaikkurichi M. Rajendran.

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Nickerson, A.J., Rajendran, V.M. Dietary Na+ depletion up-regulates NKCC1 expression and enhances electrogenic Cl− secretion in rat proximal colon. Cell. Mol. Life Sci. 80, 209 (2023). https://doi.org/10.1007/s00018-023-04857-x

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