Alterations in the intestine of Patagonian silverside (Odontesthes hatcheri) exposed to microcystin-LR: Changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity

Accumulation and toxicity of cyanobacterial toxins, particularly microcystin-LR (MCLR) have been extensively studied in fish and aquatic invertebrates. However, MCLR excretion mechanisms, which could reduce this toxin's effects, have received little attention. The Patagonian silverside, Odontes...

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Autores principales:	Bieczynski, F., Torres, W.D.C., Painefilu, J.C., Castro, J.M., Bianchi, V.A., Frontera, J.L., Paz, D.A., González, C., Martín, A., Villanueva, S.S.M., Luquet, C.M.
Formato:	JOUR
Materias:	Abcc transporters Cyanotoxin Glycoconjugates Intestinal sacs Lectin-histochemistry Multixenobiotic resistance Odontesthes hatcheri 2,4 dinitrophenyl s glutathione agglutinin calcein concanavalin A glutathione derivative glycoconjugate lectin microcystin LR multidrug resistance protein phosphoprotein phosphatase 1 unclassified drug verlukast dolichos biflorus agglutinin fluorescein derivative glutathione microcystin plant lectin propionic acid derivative quinoline derivative water pollutant cyanobacterium drug resistance ecotoxicology fish inhibition phytotoxicity protein toxin trophic structure animal experiment animal tissue Article Atherinopsidae body mass chemical composition concentration response controlled study Dolichos Dolichos biflorus enzyme activity female glycosylation histochemistry intestine cell intestine wall male nonhuman priority journal protein transport Triticum vulgaris wheat animal drug effects fluorescence microscopy intestine mucosa metabolism Smegmamorpha toxicity transport at the cellular level Argentina elongata Cyanobacteria Invertebrata Triticum aestivum Animals Biological Transport Concanavalin A Fluoresceins Glutathione Glycosylation Intestinal Mucosa Microcystins Microscopy, Fluorescence Multidrug Resistance-Associated Proteins Plant Lectins Propionates Quinolines Water Pollutants, Chemical
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_0166445X_v178_n_p106_Bieczynski
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_0166445X_v178_n_p106_Bieczynski
record_format	dspace
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Abcc transporters Cyanotoxin Glycoconjugates Intestinal sacs Lectin-histochemistry Multixenobiotic resistance Odontesthes hatcheri 2,4 dinitrophenyl s glutathione agglutinin calcein concanavalin A glutathione derivative glycoconjugate lectin microcystin LR multidrug resistance protein phosphoprotein phosphatase 1 unclassified drug verlukast dolichos biflorus agglutinin fluorescein derivative glutathione microcystin microcystin LR multidrug resistance protein plant lectin propionic acid derivative quinoline derivative water pollutant cyanobacterium drug resistance ecotoxicology fish inhibition phytotoxicity protein toxin trophic structure animal experiment animal tissue Article Atherinopsidae body mass chemical composition concentration response controlled study Dolichos Dolichos biflorus enzyme activity female glycosylation histochemistry intestine cell intestine wall male nonhuman Odontesthes hatcheri priority journal protein transport Triticum vulgaris wheat animal drug effects fluorescence microscopy glycosylation intestine mucosa metabolism Smegmamorpha toxicity transport at the cellular level water pollutant Argentina elongata Cyanobacteria Dolichos biflorus Invertebrata Odontesthes hatcheri Triticum aestivum Animals Biological Transport Concanavalin A Fluoresceins Glutathione Glycosylation Intestinal Mucosa Microcystins Microscopy, Fluorescence Multidrug Resistance-Associated Proteins Plant Lectins Propionates Quinolines Smegmamorpha Water Pollutants, Chemical
spellingShingle	Abcc transporters Cyanotoxin Glycoconjugates Intestinal sacs Lectin-histochemistry Multixenobiotic resistance Odontesthes hatcheri 2,4 dinitrophenyl s glutathione agglutinin calcein concanavalin A glutathione derivative glycoconjugate lectin microcystin LR multidrug resistance protein phosphoprotein phosphatase 1 unclassified drug verlukast dolichos biflorus agglutinin fluorescein derivative glutathione microcystin microcystin LR multidrug resistance protein plant lectin propionic acid derivative quinoline derivative water pollutant cyanobacterium drug resistance ecotoxicology fish inhibition phytotoxicity protein toxin trophic structure animal experiment animal tissue Article Atherinopsidae body mass chemical composition concentration response controlled study Dolichos Dolichos biflorus enzyme activity female glycosylation histochemistry intestine cell intestine wall male nonhuman Odontesthes hatcheri priority journal protein transport Triticum vulgaris wheat animal drug effects fluorescence microscopy glycosylation intestine mucosa metabolism Smegmamorpha toxicity transport at the cellular level water pollutant Argentina elongata Cyanobacteria Dolichos biflorus Invertebrata Odontesthes hatcheri Triticum aestivum Animals Biological Transport Concanavalin A Fluoresceins Glutathione Glycosylation Intestinal Mucosa Microcystins Microscopy, Fluorescence Multidrug Resistance-Associated Proteins Plant Lectins Propionates Quinolines Smegmamorpha Water Pollutants, Chemical Bieczynski, F. Torres, W.D.C. Painefilu, J.C. Castro, J.M. Bianchi, V.A. Frontera, J.L. Paz, D.A. González, C. Martín, A. Villanueva, S.S.M. Luquet, C.M. Alterations in the intestine of Patagonian silverside (Odontesthes hatcheri) exposed to microcystin-LR: Changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity
topic_facet	Abcc transporters Cyanotoxin Glycoconjugates Intestinal sacs Lectin-histochemistry Multixenobiotic resistance Odontesthes hatcheri 2,4 dinitrophenyl s glutathione agglutinin calcein concanavalin A glutathione derivative glycoconjugate lectin microcystin LR multidrug resistance protein phosphoprotein phosphatase 1 unclassified drug verlukast dolichos biflorus agglutinin fluorescein derivative glutathione microcystin microcystin LR multidrug resistance protein plant lectin propionic acid derivative quinoline derivative water pollutant cyanobacterium drug resistance ecotoxicology fish inhibition phytotoxicity protein toxin trophic structure animal experiment animal tissue Article Atherinopsidae body mass chemical composition concentration response controlled study Dolichos Dolichos biflorus enzyme activity female glycosylation histochemistry intestine cell intestine wall male nonhuman Odontesthes hatcheri priority journal protein transport Triticum vulgaris wheat animal drug effects fluorescence microscopy glycosylation intestine mucosa metabolism Smegmamorpha toxicity transport at the cellular level water pollutant Argentina elongata Cyanobacteria Dolichos biflorus Invertebrata Odontesthes hatcheri Triticum aestivum Animals Biological Transport Concanavalin A Fluoresceins Glutathione Glycosylation Intestinal Mucosa Microcystins Microscopy, Fluorescence Multidrug Resistance-Associated Proteins Plant Lectins Propionates Quinolines Smegmamorpha Water Pollutants, Chemical
description	Accumulation and toxicity of cyanobacterial toxins, particularly microcystin-LR (MCLR) have been extensively studied in fish and aquatic invertebrates. However, MCLR excretion mechanisms, which could reduce this toxin's effects, have received little attention. The Patagonian silverside, Odontesthes hatcheri, is an omnivorous-planktivorous edible fish, which has been shown to digest cyanobacterial cells absorbing MCLR and eliminating the toxin within 48 h without suffering significant toxic effects. We studied the effects of MCLR on glycoconjugate composition and the possible role of multidrug resistance associated proteins (Abcc) in MCLR export from the cells in O. hatcheri intestine. We treated O. hatcheri with 5 μg MCLR g−1 body mass administered with the food. Twenty four hours later, the intestines of treated and control fish were processed for lectin-histochemistry using concanavalin A (ConA), Triticum vulgaris agglutinin (WGA), and Dolichos biflorus agglutinin (DBA). MCLR affected the distribution of glycoconjugates by augmenting the proportion of ConA-positive at the expense of WGA-positive cells. We studied MCLR effects on the transport of the Abcc-like substrates 2,4-dinitrophenyl-S-glutathione (DNP-SG) and calcein in ex vivo intestine preparations (everted and no-everted sacs and strips). In treated preparations, CDNB together with MCLR (113 μg MCLR g−1 intestine, equivalent to 1.14 μmol L−1 when applied in the bath) or the Abcc inhibitor, MK571 was applied for one hour, during which DNP-SG was measured in the bath every 10 min in order to calculate mass-specific DNP-SG transport rate. MCLR significantly inhibited DNP-SG transport (p < 0.05), especially in middle intestine (47 and 24%, for luminal and serosal transport, respectively). In middle intestine strips, MCLR and MK571inhibited DNP-SG transport in a concentration dependent fashion (IC50 3.3 and 0.6 μmol L−1, respectively). In middle intestine strips incubated with calcein-AM (0.25 μmol L−1), calcein efflux was inhibited by MCLR (2.3 μmol L−1) and MK571 (3 μmol L−1) by 38 and 27%, respectively (p < 0.05). Finally, middle intestine segments were incubated with different concentrations of MCLR applied alone or together with 3 μM MK571. After one hour, protein phosphatase 1 (PP1) activity, the main target of MCLR, was measured. 2.5 μM MCLR did not produce any significant effect, while the same amount plus MK571 inhibited PP1 activity (p < 0.05). This effect was similar to that of 5 μM MCLR. Our results suggest that in O. hatcheri enterocytes MCLR is conjugated with GSH via GST and then exported to the intestinal lumen through Abcc-like transporters. This mechanism would protect the cell from MCLR toxicity, limiting toxin transport into the blood, which is probably mediated by basolateral Abccs. From an ecotoxicological point of view, elimination of MCLR through this mechanism would reduce the amount of toxin available for trophic transference. © 2016
format	JOUR
author	Bieczynski, F. Torres, W.D.C. Painefilu, J.C. Castro, J.M. Bianchi, V.A. Frontera, J.L. Paz, D.A. González, C. Martín, A. Villanueva, S.S.M. Luquet, C.M.
author_facet	Bieczynski, F. Torres, W.D.C. Painefilu, J.C. Castro, J.M. Bianchi, V.A. Frontera, J.L. Paz, D.A. González, C. Martín, A. Villanueva, S.S.M. Luquet, C.M.
author_sort	Bieczynski, F.
title	Alterations in the intestine of Patagonian silverside (Odontesthes hatcheri) exposed to microcystin-LR: Changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity
title_short	Alterations in the intestine of Patagonian silverside (Odontesthes hatcheri) exposed to microcystin-LR: Changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity
title_full	Alterations in the intestine of Patagonian silverside (Odontesthes hatcheri) exposed to microcystin-LR: Changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity
title_fullStr	Alterations in the intestine of Patagonian silverside (Odontesthes hatcheri) exposed to microcystin-LR: Changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity
title_full_unstemmed	Alterations in the intestine of Patagonian silverside (Odontesthes hatcheri) exposed to microcystin-LR: Changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity
title_sort	alterations in the intestine of patagonian silverside (odontesthes hatcheri) exposed to microcystin-lr: changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity
url	http://hdl.handle.net/20.500.12110/paper_0166445X_v178_n_p106_Bieczynski
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spelling	todo:paper_0166445X_v178_n_p106_Bieczynski2023-10-03T15:04:01Z Alterations in the intestine of Patagonian silverside (Odontesthes hatcheri) exposed to microcystin-LR: Changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity Bieczynski, F. Torres, W.D.C. Painefilu, J.C. Castro, J.M. Bianchi, V.A. Frontera, J.L. Paz, D.A. González, C. Martín, A. Villanueva, S.S.M. Luquet, C.M. Abcc transporters Cyanotoxin Glycoconjugates Intestinal sacs Lectin-histochemistry Multixenobiotic resistance Odontesthes hatcheri 2,4 dinitrophenyl s glutathione agglutinin calcein concanavalin A glutathione derivative glycoconjugate lectin microcystin LR multidrug resistance protein phosphoprotein phosphatase 1 unclassified drug verlukast dolichos biflorus agglutinin fluorescein derivative glutathione microcystin microcystin LR multidrug resistance protein plant lectin propionic acid derivative quinoline derivative water pollutant cyanobacterium drug resistance ecotoxicology fish inhibition phytotoxicity protein toxin trophic structure animal experiment animal tissue Article Atherinopsidae body mass chemical composition concentration response controlled study Dolichos Dolichos biflorus enzyme activity female glycosylation histochemistry intestine cell intestine wall male nonhuman Odontesthes hatcheri priority journal protein transport Triticum vulgaris wheat animal drug effects fluorescence microscopy glycosylation intestine mucosa metabolism Smegmamorpha toxicity transport at the cellular level water pollutant Argentina elongata Cyanobacteria Dolichos biflorus Invertebrata Odontesthes hatcheri Triticum aestivum Animals Biological Transport Concanavalin A Fluoresceins Glutathione Glycosylation Intestinal Mucosa Microcystins Microscopy, Fluorescence Multidrug Resistance-Associated Proteins Plant Lectins Propionates Quinolines Smegmamorpha Water Pollutants, Chemical Accumulation and toxicity of cyanobacterial toxins, particularly microcystin-LR (MCLR) have been extensively studied in fish and aquatic invertebrates. However, MCLR excretion mechanisms, which could reduce this toxin's effects, have received little attention. The Patagonian silverside, Odontesthes hatcheri, is an omnivorous-planktivorous edible fish, which has been shown to digest cyanobacterial cells absorbing MCLR and eliminating the toxin within 48 h without suffering significant toxic effects. We studied the effects of MCLR on glycoconjugate composition and the possible role of multidrug resistance associated proteins (Abcc) in MCLR export from the cells in O. hatcheri intestine. We treated O. hatcheri with 5 μg MCLR g−1 body mass administered with the food. Twenty four hours later, the intestines of treated and control fish were processed for lectin-histochemistry using concanavalin A (ConA), Triticum vulgaris agglutinin (WGA), and Dolichos biflorus agglutinin (DBA). MCLR affected the distribution of glycoconjugates by augmenting the proportion of ConA-positive at the expense of WGA-positive cells. We studied MCLR effects on the transport of the Abcc-like substrates 2,4-dinitrophenyl-S-glutathione (DNP-SG) and calcein in ex vivo intestine preparations (everted and no-everted sacs and strips). In treated preparations, CDNB together with MCLR (113 μg MCLR g−1 intestine, equivalent to 1.14 μmol L−1 when applied in the bath) or the Abcc inhibitor, MK571 was applied for one hour, during which DNP-SG was measured in the bath every 10 min in order to calculate mass-specific DNP-SG transport rate. MCLR significantly inhibited DNP-SG transport (p < 0.05), especially in middle intestine (47 and 24%, for luminal and serosal transport, respectively). In middle intestine strips, MCLR and MK571inhibited DNP-SG transport in a concentration dependent fashion (IC50 3.3 and 0.6 μmol L−1, respectively). In middle intestine strips incubated with calcein-AM (0.25 μmol L−1), calcein efflux was inhibited by MCLR (2.3 μmol L−1) and MK571 (3 μmol L−1) by 38 and 27%, respectively (p < 0.05). Finally, middle intestine segments were incubated with different concentrations of MCLR applied alone or together with 3 μM MK571. After one hour, protein phosphatase 1 (PP1) activity, the main target of MCLR, was measured. 2.5 μM MCLR did not produce any significant effect, while the same amount plus MK571 inhibited PP1 activity (p < 0.05). This effect was similar to that of 5 μM MCLR. Our results suggest that in O. hatcheri enterocytes MCLR is conjugated with GSH via GST and then exported to the intestinal lumen through Abcc-like transporters. This mechanism would protect the cell from MCLR toxicity, limiting toxin transport into the blood, which is probably mediated by basolateral Abccs. From an ecotoxicological point of view, elimination of MCLR through this mechanism would reduce the amount of toxin available for trophic transference. © 2016 Fil:Paz, D.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Luquet, C.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_0166445X_v178_n_p106_Bieczynski

Alterations in the intestine of Patagonian silverside (Odontesthes hatcheri) exposed to microcystin-LR: Changes in the glycosylation pattern of the intestinal wall and inhibition of multidrug resistance proteins efflux activity

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