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Physiological and molecular characterization of methotrexate transport by Malpighian tubules of adult Drosophila melanogaster.

Chahine S., O'Donnell M.J.

A radioisotope tracer technique and quantitative PCR were used to study the mechanisms and regulation of transepithelial transport of the type II organic anion methotrexate (MTX) by the Malpighian tubules of Drosophila melanogaster. Transport of MTX was saturable and Na(+)-independent; the kinetic parameters J(max) and K(t) were 437fmolmin(-1) and 23.5microM, respectively. The transport of MTX was competitively inhibited by phenol red and probenecid; non-competitively inhibited by salicylate, verapamil and MK-571; and uncompetitively inhibited by Texas Red. Dietary exposure to 0.1mM MTX led to dramatic increases in gene expression for several members of the ABC family of transporters in both the Malpighian tubules and the gut. Our results suggest that multiple transporters are upregulated in response to dietary exposure to MTX. Increased levels of the protein products which may result from expression of these genes may enhance elimination of toxic compounds such as MTX or its metabolites.

J. Insect Physiol. 55:927-935(2009) [PubMed] [Europe PMC]

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