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Characterization of glucose transport by cultured rabbit kidney proximal convoluted and proximal straight tubule cells

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Summary

Rabbit kidney proximal convoluted tubule (RPCT) and proximal straight tubule (RPST) cells were independently isolated and cultured. The kinetics of the sodium-dependent glucose transport was characterized by determining the uptake of the glucose analog alpha-methylglucopyranoside. Cell culture and assay conditions used in these experiments were based on previous experiments conducted on the renal cell line derived from the whole kidney of the Yorkshire pig (LLC-PK1). Results indicated the presence of two distinct sodium-dependent glucose transporters in rabbit renal cells: a relatively high-capacity, low-affinity transporter (Vmax=2.28±0.099 nmoles/mg protein min, Km=4.1±0.27 mM) in RPCT cells and a low-capacity, high-affinity transporter (Vmax=0.45±0.076 nmoles/mg protein min, Km=1.7±0.43 mM) in RPST cells. A relatively high-capacity, low-affinity transporter (Vmax=1.68±0.215 nmoles/mg protein min, Km=4.9±0.23 mM) was characterized in LLC-PK1 cells. Phlorizin inhibited the uptake of alpha-methylglucopyranoside in proximal convoluted, proximal straight, and LLC-PK1 cells by 90, 50, and 90%, respectively. Sodium-dependent glucose transport in all three cell types was specific for hexoses. These data are consistent with the kinetic heterogeneity of sodium-dependent glucose transport in the S1–S2 and S3 segments of the mammalian renal proximal tubule. The RPCT-RPST cultured cell model is novel, and this is the first report of sodium-dependent glucose transport characterization in primary cultures of proximal straight tubule cells. Our results support the use of cultured monolayers of RPCT and RPST cells as a model system to evaluate segment-specific differences in these renal cell types.

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Authors and Affiliations

  1. Department of Pathology, School of Medicine, University of Maryland, 21201, Baltimore, Maryland

    Pedro L. Del Valle & Anna Trifillis

  2. Lilly Research Laboratories, 46140, Greenfield, Indiana

    Charles E. Ruegg

  3. Department of Veterinary Medicine, University of Maryland, 20742, College Park, Maryland

    Andrew S. Kane

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  1. Pedro L. Del Valle
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  2. Anna Trifillis
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  3. Charles E. Ruegg
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  4. Andrew S. Kane
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Correspondence to Pedro L. Del Valle.

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Del Valle, P.L., Trifillis, A., Ruegg, C.E. et al. Characterization of glucose transport by cultured rabbit kidney proximal convoluted and proximal straight tubule cells. In Vitro Cell.Dev.Biol.-Animal 38, 218–227 (2002). https://doi.org/10.1290/1071-2690(2002)038<0218:COGTBC>2.0.CO;2

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  • DOI: https://doi.org/10.1290/1071-2690(2002)038<0218:COGTBC>2.0.CO;2

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