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Down regulation of small intestinal ion transport in PDZK1- (CAP70/NHERF3) deficient mice

  • Gastrointestinal Function
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Abstract

The PDZ-binding protein PDZK1 (CAP70/PDZ-dc-1/NHERF3) in vitro binds to cystic fibrosis transmembrane conductance regulator (CFTR), the anion exchangers SLC26A3 and SLC26A6 and the Na+/H+ exchanger NHE3, all of which are major transport proteins for intestinal anion secretion and salt absorption. This study was undertaken to search for a role of PDZK1 in regulating electrolyte transport in native murine small intestine. Short circuit current (I SC) and \({\text{HCO}}^{ - }_{3} \) secretory rate \({\left( {J_{{{\text{HCO}}^{ - }_{3} }} } \right)}\) were measured to assess electrogenic anion secretion; 22Na+ fluxes to assess sodium absorption in isolated small intestine. NHE3, CFTR, as well as NHERF1, NHERF2, and PDZK1 messenger RNA (mRNA) expression levels, and NHE3 total enterocyte and brush border membrane (BBM) protein abundance were determined by quantitative polymerase chain reaction (PCR) and Western analysis. NHE3 localization was performed by immunohistochemistry. In pdzk1 −/− jejunal mucosa, basal net Na+ absorption as well as the inhibition of Na+ absorption by forskolin was significantly reduced. In pdzk1 −/− duodenal mucosa, identical basal I SC and \(J_{{{\text{HCO}}^{ - }_{3} }} ,\) but a significant, yet mild, reduction of forskolin-stimulated Δ\(J_{{{\text{HCO}}^{ - }_{3} }} \) and ΔI SC was observed compared to +/+ tissue. Tissue conductance, morphological features, and the ΔI SC and increase in 22Na+ absorption in response to luminal glucose was identical in pdzk1 +/+ and −/− small intestine, ruling out a general absorptive defect. While CFTR mRNA expression levels were unchanged, NHE3 mRNA expression levels were significantly increased in small intestinal mucosa of pdzk1 −/− mice. Total enterocyte and BBM abundance was not significantly different, suggesting an increased NHE3 turnover, possibly due to reduced NHE3 membrane retention time. Lack of the PDZ-adapter protein PDZK1 in murine small intestine causes a mild reduction in maximal CFTR activation, but a severe defect in electroneutral Na+ absorption.

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Acknowledgment

We acknowledge the help of many colleagues during the process of establishing the methods: Prof. Christian Lytle, UC Riverside, for the advice with immunohistochemistry, and Prof. Eugene Chang and Prof. Mark Musch for the advice with Western analysis, University of Chicago. The work was supported by the DFG grants Se 460/13-1/2, DFG Se 460/17-1 and Sonderforschungsbereich SFB 621/project C9 to the USA.

Parts of this manuscript contain data prepared by Jutta Hillesheim and Mingmin Chen in fulfillment of the requirements towards their doctoral theses.

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Authors and Affiliations

  1. Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Jutta Hillesheim, Brigitte Riederer, Biguang Tuo, Mingmin Chen, Michael Manns & Ursula Seidler

  2. Department of Physiology, University of Zürich, Zurich, Switzerland

    Jürg Biber

  3. Department of Medicine, Division of Digestive Diseases, Emory School of Medicine, Atlanta, GA, USA

    Chris Yun

  4. Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

    Olivier Kocher

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  1. Jutta Hillesheim
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Correspondence to Ursula Seidler.

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Jutta Hillesheim and Brigitte Riederer contributed equally to the work.

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Hillesheim, J., Riederer, B., Tuo, B. et al. Down regulation of small intestinal ion transport in PDZK1- (CAP70/NHERF3) deficient mice. Pflugers Arch - Eur J Physiol 454, 575–586 (2007). https://doi.org/10.1007/s00424-007-0239-x

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  • DOI: https://doi.org/10.1007/s00424-007-0239-x

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