title: Similar uptake profiles of microcystin-LR and -RR in an in vitro human intestinal model.
abstract: Microcystins (MCs) are cyclic hepatotoxins produced by various species of cyanobacteria. Their structure includes two variable amino acids (AA) leading to more than 80 MC variants. In this study, we focused on the most common variant, microcystin-LR (MC-LR), and microcystin-RR (MC-RR), a variant differing by only one AA. Despite their structural similarity, MC-LR elicits higher liver toxicity than MC-RR partly due to a discrepancy in their uptake by hepatic organic anion transporters (OATP 1B1 and 1B3). However, even though ingestion is the major pathway of human exposure to MCs, intestinal absorption of MCs has been poorly addressed. Consequently, we investigated the cellular uptake of the two MC variants in the human intestinal cell line Caco-2 by immunolocalization using an anti-MC antibody. Caco-2 cells were treated for 30min to 24h with several concentrations (1-50μM) of both variants. We first confirmed the localization of OATP 3A1 and 4A1 at the cell membrane of Caco-2 cells. Our study also revealed a rapid uptake of both variants in less than 1h. The uptake profiles of the two variants did not differ in our immunostaining study neither with respect to concentration nor the time of exposure. Furthermore, we have demonstrated for the first time the nuclear localization of MC-RR and confirmed that of MC-LR. Finally, our results suggest a facilitated uptake and an active excretion of MC-LR and MC-RR in Caco-2 cells. Further investigation on the role of OATP 3A1 and 4A1 in MC uptake should be useful to clarify the mechanism of intestinal absorption of MCs and contribute in risk assessment of cyanotoxin exposure.
authors.affiliation: Anses, Fougères Laboratory, Contaminant Toxicology Unit, La Haute Marche, BP 90203, 35302 Fougères Cedex, France.
authors.forename: P
authors.initials: P
authors.lastname: Zeller
authors.forename: M
authors.initials: M
authors.lastname: Clément
authors.forename: V
authors.initials: V
authors.lastname: Fessard
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D018938:Caco-2 Cells; D000458:Cyanobacteria / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism / Q000382:microbiology; D008387:Marine Toxins; D052998:Microcystins / Q000378:metabolism*; D027361:Organic Anion Transporters / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D008387:Marine Toxins; D052998:Microcystins; D027361:Organic Anion Transporters; C479544:SLCO3A1 protein, human; C498174:SLCO4A1 protein, human; C063855:microcystin RR; C057862:cyanoginosin LR