title: Methoxychlor and Vinclozolin Induce Rapid Changes in Intercellular and Intracellular Signaling in Liver Progenitor Cells.
abstract: Methoxychlor (MXC) and vinclozolin (VIN) are well-recognized endocrine disrupting chemicals known to alter epigenetic regulations and transgenerational inheritance; however, non-endocrine disruption endpoints are also important. Thus, we determined the effects of MXC and VIN on the dysregulation of gap junctional intercellular communication (GJIC) and activation of mitogen-activated protein kinases (MAPKs) in WB-F344 rat liver epithelial cells. Both chemicals induced a rapid dysregulation of GJIC at non-cytotoxic doses, with 30 min EC50 values for GJIC inhibition being 10 µM for MXC and 126 µM for VIN. MXC inhibited GJIC for at least 24 h, while VIN effects were transient and GJIC recovered after 4 h. VIN induced rapid hyperphosphorylation and internalization of gap junction protein connexin43, and both chemicals also activated MAPK ERK1/2 and p38. Effects on GJIC were not prevented by MEK1/2 inhibitor, but by an inhibitor of phosphatidylcholine-specific phospholipase C (PC-PLC), resveratrol, and in the case of VIN, also, by a p38 inhibitor. Estrogen (ER) and androgen receptor (AR) modulators (estradiol, ICI 182,780, HPTE, testosterone, flutamide, VIN M2) did not attenuate MXC or VIN effects on GJIC. Our data also indicate that the effects were elicited by the parental compounds of MXC and VIN. Our study provides new evidence that MXC and VIN dysregulate GJIC via mechanisms involving rapid activation of PC-PLC occurring independently of ER- or AR-dependent genomic signaling. Such alterations of rapid intercellular and intracellular signaling events involved in regulations of gene expression, tissue development, function and homeostasis, could also contribute to transgenerational epigenetic effects of endocrine disruptors.
authors.affiliation: *Department of Experimental Phycology and Ecotoxicology, Institute of Botany, Brno 60200, Czech Republic; RECETOX, Faculty of Science, Masaryk University, Brno 62500, Czech Republic; Department of Pediatrics and Human Development, and Institute for Integrative Toxicology, Michigan State University, East Lansing, Michigan 48824; pavel.babica@centrum.cz.
authors.forename: Pavel
authors.initials: P
authors.lastname: Babica
authors.affiliation: Department of Pediatrics and Human Development, and Institute for Integrative Toxicology, Michigan State University, East Lansing, Michigan 48824; Department of Microbiology and Parasitology, Faculty of Medicine, National Autonomous University of Mexico, CdMx, 04510, Mexico;
authors.forename: Rimma
authors.initials: R
authors.lastname: Zurabian
authors.affiliation: Department of Pediatrics and Human Development, and Institute for Integrative Toxicology, Michigan State University, East Lansing, Michigan 48824;
authors.forename: Esha R
authors.initials: ER
authors.lastname: Kumar
authors.affiliation: Department of Pediatrics and Human Development, and Institute for Integrative Toxicology, Michigan State University, East Lansing, Michigan 48824;
authors.forename: Rajus
authors.initials: R
authors.lastname: Chopra
authors.affiliation: Department of Pediatrics and Human Development, and Institute for Integrative Toxicology, Michigan State University, East Lansing, Michigan 48824;
authors.forename: Maxwell J
authors.initials: MJ
authors.lastname: Mianecki
authors.affiliation: Department of Pediatrics and Human Development, and Institute for Integrative Toxicology, Michigan State University, East Lansing, Michigan 48824; Laboratory Animal Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu, Korea.
authors.forename: Joon-Suk
authors.initials: JS
authors.lastname: Park
authors.affiliation: *Department of Experimental Phycology and Ecotoxicology, Institute of Botany, Brno 60200, Czech Republic; RECETOX, Faculty of Science, Masaryk University, Brno 62500, Czech Republic;
authors.forename: Libor
authors.initials: L
authors.lastname: Jaša
authors.affiliation: Department of Pediatrics and Human Development, and Institute for Integrative Toxicology, Michigan State University, East Lansing, Michigan 48824;
authors.forename: James E
authors.initials: JE
authors.lastname: Trosko
authors.affiliation: Department of Pediatrics and Human Development, and Institute for Integrative Toxicology, Michigan State University, East Lansing, Michigan 48824;
authors.forename: Brad L
authors.initials: BL
authors.lastname: Upham
mesh_terms: D000818:Animals; D002450:Cell Communication / Q000187:drug effects; D002460:Cell Line; D018031:Connexin 43 / Q000378:metabolism; D017629:Gap Junctions / Q000187:drug effects; D007306:Insecticides / Q000633:toxicity*; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008731:Methoxychlor / Q000633:toxicity*; D010080:Oxazoles / Q000633:toxicity*; D051381:Rats; D011916:Rats, Inbred F344; D011944:Receptors, Androgen / Q000378:metabolism; D011960:Receptors, Estrogen / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013234:Stem Cells / Q000187:drug effects* / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D018031:Connexin 43; D007306:Insecticides; D010080:Oxazoles; D011944:Receptors, Androgen; D011960:Receptors, Estrogen; D048051:p38 Mitogen-Activated Protein Kinases; C025643:vinclozolin; D008731:Methoxychlor
keywords: endocrine disruptors; epigenetic toxicity; gap junctional intercellular communication; mitogen-activated protein kinases; non-genomic signaling.; phosphatidylcholine-specific phospholipase C