P-glycoprotein (P-gp) in the monogonont rotifer, Brachionus koreanus: Molecular characterization and expression in response to pharmaceuticals

Abstract

P-glycoprotein is involved in the efflux of diverse chemicals, including hydrophobic compounds and pharmaceuticals as a first line of defense. Here, we firstly identified and characterized the P-gp (Bk-P-gp) gene in the rotifer, Brachionus koreanus. Bk-P-gp was highly conserved in genomic organization compared to the human P-gp gene. Messenger RNA expression of Bk-P-gp revealed that it would be regulated by temperature change via 14 heat shock response elements in its promoter region. Bk-P-gp showed a high similarity of motifs/domains compared to those of vertebrates in its amino acid sequences. To check whether Bk-P-gp would be inducible, we exposed B. koreanus to six pharmaceuticals including antibiotics for use in aquaculture and observed dose- and time-dependency on transcripts of Bk-P-gp for 24 h over a wide range of concentration. Efflux assay and membrane topology supported its conserved function for transportation of a number of chemicals upon cellular damage. To reveal the effect of pharmaceuticals on the rotifer, we measured survival rate and population growth rate after exposure to six pharmaceuticals. In an acute toxicity test, both NOEC and LC50 values for all the pharmaceuticals were high for 24 h. ATP, CBZ, SMX, and TMP markedly inhibited the population growth of B. koreanus after exposure up to 100 mg/L for 10 days. In this paper, we demonstrated that various pharmaceuticals can retard growth rate with up-regulation of the P-gp gene as a cellular defense system. This finding provides a better understanding of molecular mechanisms involved in pharmaceutical-mediated cellular damage in B. koreanus.