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Molecular identification, immunolocalization, and functional activity of a vacuolar-type H+-ATPase in bovine rumen epithelium

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Abstract

In this study, we have studied the expression, localization, and functionality of vacuolar-type H+-ATPase (vH+-ATPase) and Na+/K+-ATPase in the bovine rumen epithelium. Compared with the intracellular pH (pHi) of control rumen epithelial cells (REC; 7.06 ± 0.07), application of inhibitors selective for vH+-ATPase (foliomycin) and Na+/K+-ATPase (ouabain) reduced pHi by 0.10 ± 0.03 and 0.18 ± 0.03 pH-units, respectively, thereby verifying the existence of both functional proteins. Results from qRT-PCR and immunoblotting clearly confirm the expression of vH+-ATPase B subunit in REC. However, the amount of Na+/K+-ATPase mRNA and protein is tenfold and 11-fold of those of vH+-ATPase subunit B, respectively, reflecting a lower overall abundance of the latter in REC. Na+/K+-ATPase immunostaining has revealed the protein in the plasma membrane of all REC from the stratum basale to stratum granulosum, with the highest abundance in basal cells. In contrast, the vH+-ATPase B subunit has been detected in groups of cells only, mainly localized in the stratum spinosum and stratum granulosum of the epithelium. Furthermore, vH+-ATPase has been detected in the cell membrane and in intracellular pools. Thus, functional vacuolar-type H+ pumps are expressed in REC and probably play a role in the adaptation of epithelial transport processes.

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Abbreviations

REC:

Rumen epithelial cells

vH+-ATPase:

Vacuolar-type H+-ATPase

pHi :

Intracellular pH

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Acknowledgments

We gratefully acknowledge the valuable technical assistance of K. Wolf (Institute of Veterinary Physiology, Free University Berlin) with the Western blots. We also thank R. Brose, H. Pröhl, S. Dwars and K. Marquardt (FBN Dummerstorf) for their excellent technical assistance and Dr. Theresa Jones for linguistic corrections.

This study was supported by the DFG (M. Schweigel, SCHW 652).

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Authors and Affiliations

  1. Muscle Biology and Growth, Research Institute for the Biology of Farm Animals (FBN), Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany

    Elke Albrecht

  2. Department of Veterinary Physiology, Faculty of Veterinary Medicine, Free University of Berlin, Oertzenweg 19b, 14163, Berlin, Germany

    Martin Kolisek

  3. Reproductive Biology, Research Institute for the Biology of Farm Animals (FBN), Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany

    Torsten Viergutz

  4. Research Institute of Animal Production, Slovak Agricultural Research Centre, Nitra, Komenskeho 73, 041 81, Kosice, Slovakia

    Rudolf Zitnan

  5. Research Institute for the Biology of Farm Animals (FBN), Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany

    Monika Schweigel

  6. Department of Nutritional Physiology “Oskar Kellner”, Research Institute for the Biology of Farm Animals (FBN), Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany

    Monika Schweigel

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  1. Elke Albrecht
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  2. Martin Kolisek
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  3. Torsten Viergutz
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Correspondence to Monika Schweigel.

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Communicated by G. Heldmaier.

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Albrecht, E., Kolisek, M., Viergutz, T. et al. Molecular identification, immunolocalization, and functional activity of a vacuolar-type H+-ATPase in bovine rumen epithelium. J Comp Physiol B 178, 285–295 (2008). https://doi.org/10.1007/s00360-007-0221-0

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