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Immunolocalisation of nucleoside transporters in human placental trophoblast and endothelial cells: evidence for multiple transporter isoforms

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  • Molecular and Cellular Physiology
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Abstract

Polyclonal antibodies raised against the human erythrocyte nucleoside transporter were used to investigate the distribution of the nucleoside transporters in the placenta. Immunoblots of brush-border membranes isolated from the human syncytiotrophoblast revealed a cross-reactive species that co-migrated with the erythrocyte nucleoside transporter as a broad band of apparent M r 55,000. In contrast, no labelling was detected in basal membranes containing a similar number of equilibrative nucleoside transporters as assessed by nitrobenzylthioinosine (NBMPR)-binding. The absence of cross-reactive epitopes in basal membranes and their presence in brush-border membranes was confirmed by confocal immunofluorescence microscopy. These results suggest that at least two isoforms of the NBMPR-sensitive nucleoside transporter are present in the human placenta. The lumenal surfaces of fetal capillaries, small placental vessels and umbilical vein were also strongly labelled by the antibody, a finding that suggests that the high fetal-placental adenosine uptake previously reported is due to endothelial transporters.

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Abbreviations

NT :

The human erythrocyte nucleoside transporter

Anti-NT :

Polyclonal antibody against the nucleoside transporter

NBMPR :

Nitrobenzylthioinosine {6-[(4-nitrobenzyl)thio]-9-β-D-ribofuranosylpurine}.

GLUTl :

The human erythrocyte glucose transporter

PBS :

Phosphate-buffered saline

TBS :

TRIS-buffered saline

TTBS :

TRIS-buffered saline containing 0.2% Tween-20

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Authors and Affiliations

  1. Departmento of Fisiología y Biofísica, Universidad de Chile, Casilla, 70005, Santiago 7, Chile

    L. F. Barros & D. L. Yudilevich

  2. Department of Biochemistry and Molecular Biology, University of Leeds, LS2 9JT, Leeds, UK

    L. F. Barros, N. Beaumont & S. A. Baldwin

  3. Research School of Biosciences, University of Kent, CT2 7NJ, Canterbury, Kent, UK

    S. M. Jarvis

  4. Department of Physiology, University of Alberta, T6G 2H7, Edmonton, Canada

    J. D. Young

Authors
  1. L. F. Barros
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  2. D. L. Yudilevich
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  3. S. M. Jarvis
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  4. N. Beaumont
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  5. J. D. Young
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  6. S. A. Baldwin
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Barros, L.F., Yudilevich, D.L., Jarvis, S.M. et al. Immunolocalisation of nucleoside transporters in human placental trophoblast and endothelial cells: evidence for multiple transporter isoforms. Pflugers Arch. 429, 394–399 (1995). https://doi.org/10.1007/BF00374155

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  • DOI: https://doi.org/10.1007/BF00374155

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