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Na-dependent D-Glucose Transport by Intestinal Brush Border Membrane Vesicles from Gilthead Sea Bream (Sparus aurata)

Abstract

Brush border membrane vesicles (BBMV) enriched in sucrase, maltase and alkaline phosphatase, and impoverished in Na+-K+-ATPase, were isolated from proximal and distal intestine of the gilthead sea bream (Sparus aurata) by a MgCl2 precipitation method. Vesicles were suitable for the study of the characteristics of D-glucose apical transport. Only one D-glucose carrier was found in vesicles from each intestinal segment. In both cases, the D-glucose transport system was sodium-dependent, phlorizin-sensitive, significantly inhibited by D-glucose, D-galactose, α-methyl-D-glucose, 3-O-methyl-D-glucose and 2-deoxy-D-glucose, and showed stereospecificity. Apparent affinity constants of D-glucose transport (Kt) were 0.24  ±  0.03 mM in proximal and 0.18  ±  0.03 mM in distal intestine. Maximal rate of influx (Jmax) was 47.3  ±  2.2 pmols. mg−1 protein for proximal and 27.3  ±  3.6 pmols. mg−1 protein for distal intestine. Specific phlorizin binding and relative abundance of an anti-SGLT1 reactive protein were significantly higher in proximal than in distal BBMV. These results suggest the presence of the same D-glucose transporter along the intestine, with a higher density in the proximal portion. This transporter is compatible with the sodium-dependent D-glucose carrier described for other fish and with the SGLT1 of higher vertebrates.

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Acknowledgments

This work has been supported by Spanish government (grant MAR97-0408-00-02 of CICYT) and Generalitat de Catalunya (grant 2001-SGR-00142 of CIRIT, and Centre de Referència en Aqüicultura). M. Sala-Rabanal was a recipient of a Formació de Personal Investigador grant from the Generalitat de Catalunya. The authors thank Dr. Carles Garriga and Anna Pérez-Bosque for their assistance in the performance of the Western blot and the immunohistochemichal analyses, respectively.

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Correspondence to M.A. Gallardo.

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Sala-Rabanal, M., Gallardo, M., Sánchez, J. et al. Na-dependent D-Glucose Transport by Intestinal Brush Border Membrane Vesicles from Gilthead Sea Bream (Sparus aurata). J Membrane Biol 201, 85–96 (2004). https://doi.org/10.1007/s00232-004-0710-y

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  • DOI: https://doi.org/10.1007/s00232-004-0710-y

Keywords

  • SGLT1
  • Proximal intestine
  • Distal intestine
  • Brush border membrane vesicles
  • Freezing
  • Fish