Effects of microcystin-LR on the expression of P-glycoprotein in Jenynsia multidentata.
The multixenobiotic resistance phenomenon (MXR) related to the P-glycoprotein multidrug transporter protein (P-gp) has been identified and characterized in several aquatic organisms. In the present work, we prove the presence of a P-gp in liver, gills and brain of Jenynsia multidentata by Western Blot and RT-PCR. A 170 kDa protein has been found in liver and gills while in brain a approximately 80 kDa protein has been detected. The partial nucleotide sequence obtained in this autochthonous fish showed high similarity ranging from 83% to 92% with other fishes. In addition, P-gp expression in this fish was evaluated after time and dose-dependent exposures to the cyanotoxin microcystin-LR. Individuals were exposed to MC-LR at concentrations of 2, 5 and 10 microg L(-1) for 24h and for 6, 12 and 24h at 2 microg L(-1) MC-LR. Changes in P-gp expression were observed in liver, gills and brain. However, this response was tissue specific. Only in gills of J. multidentata P-gp expression, measured either by real-time RT-PCR or Western Blot, was significantly higher compared to controls at most tested times and doses. A 3-fold increase with respect to controls was found at 12h by RT-PCR and after 24h by Western Blot. In dose-dependent experiments the maximum P-gp expression was observed at 2 microg L(-1) MC-LR, measured by both RT-PCR and Western Blot. In the liver, P-gp protein levels were significantly increased after 24h of exposure, at every toxin dose tested. Thus, probably longer exposures would show also significant increases in this tissue. Considering these results we can propose that P-gp belongs to the defence system involved in the response to MC-LR in J. multidentata.
Amé
María Valeria
MV
Universidad Nacional de Córdoba-CONICET, Facultad Ciencias Químicas, Dto. Bioquímica Clínica-CIBICI, Haya de la Torre esq. Medina Allende, Ciudad Universitaria, 5000-Córdoba, Argentina. vame@mail.fcq.unc.edu.ar
Baroni
María Verónica
MV
Galanti
Lucas Nicolás
LN
Bocco
José Luis
JL
Wunderlin
Daniel Alberto
DA
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D003532:Cyprinodontiformes / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects*; D005880:Gills / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008387:Marine Toxins; D052998:Microcystins / Q000633:toxicity*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D008387:Marine Toxins; D052998:Microcystins; C057862:cyanoginosin LR