Mussel blood cells, resistant to the cytotoxic effects of okadaic acid, do not express cell membrane p-glycoprotein activity (multixenobiotic resistance).
Okadaic acid (OA) is a dinoflagellate toxin, accumulating in shellfish and causing diarrhetic shellfish poisoning (DSP) in humans. OA is a highly cytotoxic agent in most cell lines because of its inhibiting properties of protein phosphatases. So far, the cytotoxicity of OA in mussels, the main vectors of DSP, has not been investigated. In this paper, the viability of mussel (Mytilus edulis) blood cells incubated in 10 nM-1 microM OA was studied. After 72 h of exposure, viability was reduced to 54% in 1 microM OA compared with 88% in control cells. This yielded a LC50 of >1 microM for OA, which is 30-1000-times higher compared with other cell types. It was hypothesised that P-glycoprotein (p-gp) activity (multixenobiotic resistance, MXR) contributed to the resistance to OA. Vincristine and rhodamine B was used as p-gp substrates and verapamil or staurosporine (ST) as inhibitors of p-gp transport. However, no indications of cell membrane p-gp activity were detected. Instead, experimental observations led to the conclusion that a MXR transport system was present within lysosomal membranes. Various concentrations of OA did not affect the dynamics of vincristine in blood cells. As a positive control for the assay, p-gp activity was measured in mussel gill tissue. The efflux of rhodamine B was reduced by verapamil, which is, considered evidence for cell membrane p-gp activity, thus the accuracy of the method was confirmed. Rhodamine B efflux was also reduced by OA in gill tissue, which suggested that OA is either a competitive substrate or inhibitor of p-gp activity. When the volume of the lysosomal compartment was measured in blood cells pre-exposed to OA, a significant increase was detected compared with control cells. It was proposed that uptake and storage of OA within the lysosomal system might protect mussel blood cells from the cytotoxic effects of this compound.
Svensson
Susanne
S
Department of Zoology/Zoophysiology, Göteborg University, Box 463, SE-405 30 Göteborg, Sweden. susanne.svensson@zool.gu.se
Särngren
Anders
A
Förlin
Lars
L
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis*; D000704:Analysis of Variance; D000818:Animals; D001681:Biological Assay; D049872:Bivalvia / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D018432:Drug Resistance, Multiple; D006434:Hemocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism; D019319:Okadaic Acid / Q000633:toxicity*; D012235:Rhodamines; D014750:Vincristine; D015262:Xenobiotics
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D012235:Rhodamines; D015262:Xenobiotics; D019319:Okadaic Acid; D014750:Vincristine; C029773:rhodamine B