Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8552858 | Toxicology | 2018 | 10 Pages |
Abstract
Methylmercury (MeHg) is an environmental neurotoxicant that inhibits neuronal migration. This process requires several cyclic steps involving the formation of membrane protrusions (lamellipodia and filopodia) and focal adhesion turnover. FAK and Src are critical proteins that regulate both processes. The FAK-Src complex promotes the activation of Rac1 and Cdc42, two GTPases involved in the remodeling of the actin cytoskeletal network. Here, we studied the effect of MeHg (1, 10, 100, 500 and 1000Â nM) on cell migration, the formation of cell protrusions, focal adhesion location and the activation of FAK, Src, Rac1 and Cdc42 using the SH-SY5Y neuroblastoma cell line stimulated with PDGF-BB (PDGF). The data show that MeHg (1-500Â nM) inhibited PDGF-stimulated cell migration. In PDGF-stimulated cells, MeHg (100-1000Â nM) decreased protrusions and increased the size of the p-FAKY397 clusters. MeHg also inhibited PDGF-induced FAK and Src activation and, at 100Â nM, MeHg inhibited the activation of Rac1 and Cdc42. Altogether, the findings show that low concentrations of MeHg inhibit SH-SY5Y cell migration by disrupting the activation and disassembly of FAK. This negatively affects the activation of Src, Rac1 and Cdc42, all of which are critical proteins for the regulation of cell movement. These effects could be related to the MeHg-mediated inhibition of PDGF-induced formation of lamellipodia and filopodia, focal adhesion disassembly and PDGF-induced movement.
Keywords
MeHgHRPDock180RhoASRCEC50Platelet-derived growth factor BBCysSHP-2PAGEFAKCTCFNGFGEFRac1PDGFRCrkCDC42PDGF4.1 ezrin radixin moesinTRITC-phalloidinEGFRELMO1Proto-oncogene tyrosine-protein kinase SrcSDSFBSPTP-PESTFERMPAXPVDFPNDPaxillinBDNFBSACa2+DMSOMTTp130Casbovine serum albuminpolyacrylamide gel electrophoresisRIPA bufferradioimmunoprecipitation assay bufferanalysis of varianceANOVADimethyl sulfoxidegestation daypostnatal daysodium dodecyl sulfatefetal bovine serumCysteinebrain derived neurotrophic factorRas homolog gene family member Aguanine nucleotide exchange factornerve growth factorPKLcorrected total cell fluorescenceMethylmercuryCell migrationhalf maximal effective concentrationHorseradish peroxidasecell division control protein 42 homologplatelet-derived growth factor receptor βPolyvinylidene fluorideFocal adhesionsCalciumfocal adhesion kinaseEpidermal growth factor receptor
Related Topics
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Authors
A. José Alberto Hernández, V. Liliana Reyes, Damaris Albores-GarcÃa, RocÃo Gómez, Emma S. Calderón-Aranda,