The multi-xenobiotic resistance (MXR) efflux activity in hemocytes of Mytilus edulis is mediated by an ATP binding cassette transporter of class C (ABCC) principally inducible in eosinophilic granulocytes
Identifieur interne : 000345 ( France/Analysis ); précédent : 000344; suivant : 000346The multi-xenobiotic resistance (MXR) efflux activity in hemocytes of Mytilus edulis is mediated by an ATP binding cassette transporter of class C (ABCC) principally inducible in eosinophilic granulocytes
Auteurs : Damien Rioult [France] ; Jennifer Pasquier [France] ; Céline Boulange-Lecomte [France] ; Agnès Poret [France] ; Imane Abbas [Liban] ; Matthieu Marin [France] ; Christophe Minier [France] ; Frank Le Foll [France]Source :
- Aquatic toxicology [ 0166-445X ] ; 2014.
Descripteurs français
- Pascal (Inist)
- Wicri :
- geographic : France.
- topic : Migration, Milieu marin, Polluant.
English descriptors
- KwdEn :
Abstract
In marine and estuarine species, immunotoxic and/or immunomodulatory mechanisms are the crossroad of interactions between xenobiotics, microorganisms and physicochemical variations of the environment. In mussels, immunity relies exclusively on innate responses carried out by cells collectively called hemocytes and found in the open hemolymphatic circulatory system of these organisms. However, hemocytes do not form a homogenous population of immune cells since distinct subtypes of mussel blood cells can be distinguished by cytochemistry, flow cytometry or cell motility analysis. Previous studies have also shown that these cells are able to efflux xenobiotics by means of ATP binding cassette (ABC) transporter activities conferring a multixenobiotic resistance (MXR) phenotype. ABC transporters corresponding to vertebrate class B/P-glycoprotein (P-gp) and to class C/multidrug resistance related protein (MRP) are characterized in Mytilidae. Herein, we have investigated the relative contributions of ABCB- and ABCC-mediated efflux within the different hemocyte subpopulations of Mytilus edulis mussels, collected from areas differentially impacted by chemical contaminants in Normandy (France). RT-PCR analyses provide evidence for the presence of ABCB and ABCC transporters transcripts in hemocytes. Immunodetection of ABCB/P-gp with the monoclonal antibody UIC2 in living hemocytes revealed that expression was restricted to granular structures of spread cells. Efflux transporter activities, with calcein-AM as fluorescent probe, were measured by combining flow cytometry to accurate Coulter cell size measurements in order to get a cell-volume normalized fluorescence concentration. In these conditions, basal fluorescence levels were higher in hemocytes originating from Yport (control site) than in cells collected from the harbor of Le Havre, where mussels are more exposed to with persistent pollutants. By using specific ABCB/P-gp (verapamil, PSC833, zosuquidar) and ABCC/MRP (MK571) blockers, we show that MXR activity is only carried out by MRP-type transporters in M. edulis hemocytes. In addition, cell-type-gated flow cytometry and calculation of the MXR activity factor indicate that ABCC-efflux activity is higher and more inducible in eosinophilic granulocytes than in other hemocyte subtypes. We conclude that, in the hemocytes of M. edulis, MXR phenotype is mediated by an ABCC/MRP-type transporter activity principally supported by eosinophilic granulocytes. A role for ABC transporters in hemocyte migration is discussed.
Affiliations:
- France, Liban
- Haute-Normandie, Hauts-de-France, Nord-Pas-de-Calais, Région Normandie
- Le Havre, Villeneuve d'Ascq
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SN3, Université des Sciences et Technologies de Lille</s1><s2>59655 Villeneuve d'Ascq</s2><s3>FRA</s3><sZ>6 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Hauts-de-France</region><region type="old region" nuts="2">Nord-Pas-de-Calais</region><settlement type="city">Villeneuve d'Ascq</settlement></placeName></affiliation></author><author><name sortKey="Minier, Christophe" sort="Minier, Christophe" uniqKey="Minier C" first="Christophe" last="Minier">Christophe Minier</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Ecotoxicology, UPRES EA 3222, IFRMP23, University of Le Havre</s1><s2>76058 Le Havre</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Région Normandie</region><region type="old region" nuts="2">Haute-Normandie</region><settlement type="city">Le Havre</settlement></placeName></affiliation></author><author><name sortKey="Le Foll, Frank" sort="Le Foll, Frank" uniqKey="Le Foll F" first="Frank" last="Le Foll">Frank Le Foll</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Ecotoxicology, UPRES EA 3222, IFRMP23, University of Le Havre</s1><s2>76058 Le Havre</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Région Normandie</region><region type="old region" nuts="2">Haute-Normandie</region><settlement type="city">Le Havre</settlement></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">14-0185695</idno><date when="2014">2014</date><idno type="stanalyst">PASCAL 14-0185695 INIST</idno><idno type="RBID">Pascal:14-0185695</idno><idno type="wicri:Area/PascalFrancis/Corpus">000008</idno><idno 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Havre</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Région Normandie</region><region type="old region" nuts="2">Haute-Normandie</region><settlement type="city">Le Havre</settlement></placeName></affiliation></author><author><name sortKey="Pasquier, Jennifer" sort="Pasquier, Jennifer" uniqKey="Pasquier J" first="Jennifer" last="Pasquier">Jennifer Pasquier</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Ecotoxicology, UPRES EA 3222, IFRMP23, University of Le Havre</s1><s2>76058 Le Havre</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Région Normandie</region><region type="old region" nuts="2">Haute-Normandie</region><settlement type="city">Le Havre</settlement></placeName></affiliation></author><author><name sortKey="Boulange Lecomte, Celine" sort="Boulange Lecomte, Celine" uniqKey="Boulange Lecomte C" first="Céline" last="Boulange-Lecomte">Céline Boulange-Lecomte</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Ecotoxicology, UPRES EA 3222, IFRMP23, University of Le Havre</s1><s2>76058 Le Havre</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Région Normandie</region><region type="old region" nuts="2">Haute-Normandie</region><settlement type="city">Le Havre</settlement></placeName></affiliation></author><author><name sortKey="Poret, Agnes" sort="Poret, Agnes" uniqKey="Poret A" first="Agnès" last="Poret">Agnès Poret</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Ecotoxicology, UPRES EA 3222, IFRMP23, University of Le Havre</s1><s2>76058 Le Havre</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Région Normandie</region><region type="old region" nuts="2">Haute-Normandie</region><settlement type="city">Le Havre</settlement></placeName></affiliation></author><author><name sortKey="Abbas, Imane" sort="Abbas, Imane" uniqKey="Abbas I" first="Imane" last="Abbas">Imane Abbas</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Research and Development Department, Lebanese Atomic Energy Commission - CNRS</s1><s2>Beirut</s2><s3>LBN</s3><sZ>5 aut.</sZ></inist:fA14><country>Liban</country><wicri:noRegion>Beirut</wicri:noRegion></affiliation></author><author><name sortKey="Marin, Matthieu" sort="Marin, Matthieu" uniqKey="Marin M" first="Matthieu" last="Marin">Matthieu Marin</name><affiliation wicri:level="3"><inist:fA14 i1="03"><s1>Laboratoire de Régulation des Signaux de Division, EA 4020, IFR 147, Bât. SN3, Université des Sciences et Technologies de Lille</s1><s2>59655 Villeneuve d'Ascq</s2><s3>FRA</s3><sZ>6 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Hauts-de-France</region><region type="old region" nuts="2">Nord-Pas-de-Calais</region><settlement type="city">Villeneuve d'Ascq</settlement></placeName></affiliation></author><author><name sortKey="Minier, Christophe" sort="Minier, Christophe" uniqKey="Minier C" first="Christophe" last="Minier">Christophe Minier</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Ecotoxicology, UPRES EA 3222, IFRMP23, University of Le Havre</s1><s2>76058 Le Havre</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Région Normandie</region><region type="old region" nuts="2">Haute-Normandie</region><settlement type="city">Le Havre</settlement></placeName></affiliation></author><author><name sortKey="Le Foll, Frank" sort="Le Foll, Frank" uniqKey="Le Foll F" first="Frank" last="Le Foll">Frank Le Foll</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Ecotoxicology, UPRES EA 3222, IFRMP23, University of Le Havre</s1><s2>76058 Le Havre</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region" nuts="2">Région Normandie</region><region type="old region" nuts="2">Haute-Normandie</region><settlement type="city">Le Havre</settlement></placeName></affiliation></author></analytic><series><title level="j" type="main">Aquatic toxicology</title><title level="j" type="abbreviated">Aquat. toxicol.</title><idno type="ISSN">0166-445X</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Aquatic toxicology</title><title level="j" type="abbreviated">Aquat. toxicol.</title><idno type="ISSN">0166-445X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>ABC transporter</term><term>Carrier protein</term><term>Defense mechanism</term><term>Detoxification</term><term>Dose activity relation</term><term>Edible mollusc</term><term>Flow cytometry</term><term>France</term><term>Granulocyte</term><term>Hemocyte</term><term>Leukocyte</term><term>Marine environment</term><term>Migration</term><term>Mussel</term><term>Mytilus edulis</term><term>Natural immunity</term><term>Pollutant</term><term>Xenobiotic</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Xénobiotique</term><term>Mécanisme défense</term><term>Hémocyte</term><term>Mytilus edulis</term><term>Transporteur ABC</term><term>Granulocyte</term><term>Moule (comestible)</term><term>France</term><term>Immunité naturelle</term><term>Cytométrie flux</term><term>Migration</term><term>Milieu marin</term><term>Détoxication</term><term>Relation dose réponse</term><term>Polluant</term><term>Protéine transport</term><term>Leucocyte</term><term>Mollusque comestible</term></keywords><keywords scheme="Wicri" type="geographic" xml:lang="fr"><term>France</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Migration</term><term>Milieu marin</term><term>Polluant</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">In marine and estuarine species, immunotoxic and/or immunomodulatory mechanisms are the crossroad of interactions between xenobiotics, microorganisms and physicochemical variations of the environment. In mussels, immunity relies exclusively on innate responses carried out by cells collectively called hemocytes and found in the open hemolymphatic circulatory system of these organisms. However, hemocytes do not form a homogenous population of immune cells since distinct subtypes of mussel blood cells can be distinguished by cytochemistry, flow cytometry or cell motility analysis. Previous studies have also shown that these cells are able to efflux xenobiotics by means of ATP binding cassette (ABC) transporter activities conferring a multixenobiotic resistance (MXR) phenotype. ABC transporters corresponding to vertebrate class B/P-glycoprotein (P-gp) and to class C/multidrug resistance related protein (MRP) are characterized in Mytilidae. Herein, we have investigated the relative contributions of ABCB- and ABCC-mediated efflux within the different hemocyte subpopulations of Mytilus edulis mussels, collected from areas differentially impacted by chemical contaminants in Normandy (France). RT-PCR analyses provide evidence for the presence of ABCB and ABCC transporters transcripts in hemocytes. Immunodetection of ABCB/P-gp with the monoclonal antibody UIC2 in living hemocytes revealed that expression was restricted to granular structures of spread cells. Efflux transporter activities, with calcein-AM as fluorescent probe, were measured by combining flow cytometry to accurate Coulter cell size measurements in order to get a cell-volume normalized fluorescence concentration. In these conditions, basal fluorescence levels were higher in hemocytes originating from Yport (control site) than in cells collected from the harbor of Le Havre, where mussels are more exposed to with persistent pollutants. By using specific ABCB/P-gp (verapamil, PSC833, zosuquidar) and ABCC/MRP (MK571) blockers, we show that MXR activity is only carried out by MRP-type transporters in M. edulis hemocytes. In addition, cell-type-gated flow cytometry and calculation of the MXR activity factor indicate that ABCC-efflux activity is higher and more inducible in eosinophilic granulocytes than in other hemocyte subtypes. We conclude that, in the hemocytes of M. edulis, MXR phenotype is mediated by an ABCC/MRP-type transporter activity principally supported by eosinophilic granulocytes. A role for ABC transporters in hemocyte migration is discussed.</div></front></TEI><affiliations><list><country><li>France</li><li>Liban</li></country><region><li>Haute-Normandie</li><li>Hauts-de-France</li><li>Nord-Pas-de-Calais</li><li>Région Normandie</li></region><settlement><li>Le Havre</li><li>Villeneuve d'Ascq</li></settlement></list><tree><country name="France"><region name="Région Normandie"><name sortKey="Rioult, Damien" sort="Rioult, Damien" uniqKey="Rioult D" first="Damien" last="Rioult">Damien Rioult</name></region><name sortKey="Boulange Lecomte, Celine" sort="Boulange Lecomte, Celine" uniqKey="Boulange Lecomte C" first="Céline" last="Boulange-Lecomte">Céline Boulange-Lecomte</name><name sortKey="Le Foll, Frank" sort="Le Foll, Frank" uniqKey="Le Foll F" first="Frank" last="Le Foll">Frank Le Foll</name><name sortKey="Marin, Matthieu" sort="Marin, Matthieu" uniqKey="Marin M" first="Matthieu" last="Marin">Matthieu Marin</name><name sortKey="Minier, Christophe" sort="Minier, Christophe" uniqKey="Minier C" first="Christophe" last="Minier">Christophe Minier</name><name sortKey="Pasquier, Jennifer" sort="Pasquier, Jennifer" uniqKey="Pasquier J" first="Jennifer" last="Pasquier">Jennifer Pasquier</name><name sortKey="Poret, Agnes" sort="Poret, Agnes" uniqKey="Poret A" first="Agnès" last="Poret">Agnès Poret</name></country><country name="Liban"><noRegion><name sortKey="Abbas, Imane" sort="Abbas, Imane" uniqKey="Abbas I" first="Imane" last="Abbas">Imane Abbas</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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