title: Ethanol Attenuates Histiotrophic Nutrition Pathways and Alters the Intracellular Redox Environment and Thiol Proteome during Rat Organogenesis.
abstract: Ethanol (EtOH) is a reactive oxygen-generating teratogen involved in the etiology of structural and functional developmental defects. Embryonic nutrition, redox environment, and changes in the thiol proteome following EtOH exposures (1.56.0 mg/ml) were studied in rat whole embryo culture. Glutathione (GSH) and cysteine (Cys) concentrations with their respective intracellular redox potentials (Eh) were determined using high-performance liquid chromatography. EtOH reduced GSH and Cys concentrations in embryo (EMB) and visceral yolk sac (VYS) tissues, and also in yolk sac and amniotic fluids. These changes produced greater oxidation as indicated by increasingly positive Eh values. EtOH reduced histiotrophic nutrition pathway activities as measured by the clearance of fluorescin isothiocyanate (FITC)-albumin from culture media. A significant decrease in total FITC clearance was observed at all concentrations, reaching approximately 50% at the highest dose. EtOH-induced changes to the thiol proteome were measured in EMBs and VYSs using isotope-coded affinity tags. Decreased concentrations for specific proteins from cytoskeletal dynamics and endocytosis pathways (α-actinin, α-tubulin, cubilin, and actin-related protein 2); nuclear translocation (Ran and RanBP1); and maintenance of receptor-mediated endocytosis (cubilin) were observed. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis also identified a decrease in ribosomal proteins in both EMB and VYS. Results show that EtOH interferes with nutrient uptake to reduce availability of amino acids and micronutrients required by the conceptus. Intracellular antioxidants such as GSH and Cys are depleted following EtOH and Eh values increase. Thiol proteome analysis in the EMB and VYS show selectively altered actin/cytoskeleton, endocytosis, ribosome biogenesis and function, nuclear transport, and stress-related responses.
authors.affiliation: *Department of Environmental Health Sciences, University of Michigan, Ann Arbor, Michigan 48109;
authors.forename: Joseph L
authors.initials: JL
authors.lastname: Jilek
authors.affiliation: *Department of Environmental Health Sciences, University of Michigan, Ann Arbor, Michigan 48109;
authors.forename: Karilyn E
authors.initials: KE
authors.lastname: Sant
authors.affiliation: *Department of Environmental Health Sciences, University of Michigan, Ann Arbor, Michigan 48109;
authors.forename: Katherine H
authors.initials: KH
authors.lastname: Cho
authors.affiliation: Biotechnology Core Facility Branch, Centers for Disease Control, Atlanta, Georgia 30333; and.
authors.forename: Matthew S
authors.initials: MS
authors.lastname: Reed
authors.affiliation: Biotechnology Core Facility Branch, Centers for Disease Control, Atlanta, Georgia 30333; and.
authors.forename: Jan
authors.initials: J
authors.lastname: Pohl
authors.affiliation: Department of Physiology and Developmental Biology, College of Life Sciences, Brigham Young University, Provo, Utah 84602.
authors.forename: Jason M
authors.initials: JM
authors.lastname: Hansen
authors.affiliation: *Department of Environmental Health Sciences, University of Michigan, Ann Arbor, Michigan 48109; charris@umich.edu.
authors.forename: Craig
authors.initials: C
authors.lastname: Harris
mesh_terms: D000818:Animals; D003545:Cysteine / Q000032:analysis; D000431:Ethanol / Q000633:toxicity*; D005260:Female; D047109:Fetal Development / Q000187:drug effects; D005333:Fetus / Q000737:chemistry / Q000187:drug effects; D005978:Glutathione / Q000032:analysis; D008297:Male; D053858:Metabolic Networks and Pathways / Q000187:drug effects*; D038081:Organogenesis / Q000187:drug effects* / Q000502:physiology; D010084:Oxidation-Reduction / Q000187:drug effects*; D011247:Pregnancy; D020543:Proteome / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D020543:Proteome; D013438:Sulfhydryl Compounds; D000431:Ethanol; D005978:Glutathione; D003545:Cysteine
keywords: cysteine; embryo; ethanol; glutathione; histiotrophic nutrition; organogenesis; redox environment; redox potential; thiol proteome; visceral yolk sac