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Immunolocalisation of an ABC transporter, P-glycoprotein, in the eggshells and cuticles of free-living and parasitic stages of Haemonchus contortus

Abstract

Recent data have suggested that P-glycoprotein (Pgp), working as membrane efflux “pumps”, plays a major role in the transport of anthelmintic drugs in parasitic nematodes of ruminants. Flow cytometry analyses has shown that active Pgp is probably present in the external layers of Haemonchus contortus eggshells, following staining with the mouse monoclonal anti-human MDR1 antibody UIC2, which binds to Pgp in its active conformation. We evaluated the presence and distribution of this protein in the envelopes (eggshells and cuticles) of H. contortus and compared the various stages (eggs, L1–L2 larvae, L3 larvae, adult male and female worms). Electrophoresis revealed a 170-kDa band, corresponding to the molecular weight of Pgp in all stages. Indirect immunofluorescence staining with UIC2 showed Pgp to be located in the external layer of eggshells or cuticles. Transmission electron microscopy was used to localise Pgp more accurately in the three layers of the eggshells and cuticles. The conformation and biological functions of this protein, which we did not expect to find in such structures, remain to be determined.

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Acknowledgements

We thank Dr Yan Van Wyk, Dr Gilles Aumont and Professor Roger Prichard for kindly providing H. contortus isolates, Dr Laurence Guilloteau for helping with immunological staining experiments, Mr. Limouzin and his team for the careful maintenance of animals and Marie-Colette Fauré and her team for their invaluable help in tracking down references. We also thank the “Region Centre” for financing this research.

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Correspondence to Dominique Kerboeuf.

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Riou, M., Koch, C., Delaleu, B. et al. Immunolocalisation of an ABC transporter, P-glycoprotein, in the eggshells and cuticles of free-living and parasitic stages of Haemonchus contortus. Parasitol Res 96, 142–148 (2005). https://doi.org/10.1007/s00436-005-1345-3

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Keywords

  • Adult Worm
  • Ivermectin
  • Intestinal Cell
  • Parasitic Stage
  • Female Worm