title: Homocysteine-induced changes in cell proliferation and differentiation in the chick embryo spinal cord: implications for mechanisms of neural tube defects (NTD).
abstract: Maternal hyperhomocysteinemia during pregnancy is associated with increased risk of NTD in the offspring. Our study investigated the effects of homocysteine (Hcy) on proliferation and neuronal differentiation of the spinal cord cells in a chick embryo model. Embryos were treated with 20μmol D-L Hcy/50μL saline solution at embryonic day 2 (E2) and analyzed at embryonic days 4 (E4) and 6 (E6). Control embryos received exclusively 50μL saline solution. We performed immunolocalization and flow cytometry analyses using antibodies anti-phosphohistone H3 (pH3), anti-proliferating cell nuclear antigen (PCNA), anti-β-tubulin III and anti-p53. Our results revealed that Hcy interferes in the proliferation of the neural cells, and that this effect is age-dependent and differed between Hcy-treated embryos with and without NTD. Also, Hcy induced a decrease of neuronal differentiation in the spinal cord at both embryonic ages. These findings contribute to clarifying the cellular bases of NTD genesis, under experimental hiperhomocysteinemia.
authors.affiliation: Departamento de Fisioterapia, Centro de Ciências da Saúde e do Esporte, UDESC, Rua Pascoal Simone 358, Coqueiros, Florianópolis, SC, 88080-350, Brazil; Centro Universitário Estácio de Sá Santa Catarina, Avenida Leoberto Leal 431, São José, SC, 88117-001, Brazil. Electronic address: karoline.bianchini@udesc.br.
authors.forename: Karoline
authors.initials: K
authors.lastname: Kobus-Bianchini
authors.affiliation: Programa de Pós-graduação em Biologia Celular e do Desenvolvimento, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, Florianópolis, SC, 88040-900, Brazil.
authors.forename: Gilian Fernando
authors.initials: GF
authors.lastname: Bourckhardt
authors.affiliation: Centro Universitário Católica de Santa Catarina, Rua Visconde de Taunay 427, Centro, Joinville, SC, 89203-005, Brazil.
authors.forename: Dib
authors.initials: D
authors.lastname: Ammar
authors.affiliation: Programa de Pós-graduação em Biologia Celular e do Desenvolvimento, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, Florianópolis, SC, 88040-900, Brazil; Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, Florianópolis, SC, 88040-900, Brazil.
authors.forename: Evelise Maria
authors.initials: EM
authors.lastname: Nazari
authors.affiliation: Programa de Pós-graduação em Biologia Celular e do Desenvolvimento, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, Florianópolis, SC, 88040-900, Brazil; Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, Florianópolis, SC, 88040-900, Brazil.
authors.forename: Yara Maria Rauh
authors.initials: YMR
authors.lastname: Müller
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002642:Chick Embryo; D006657:Histones / Q000378:metabolism; D006710:Homocysteine / Q000633:toxicity*; D009436:Neural Tube Defects; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology / Q000187:drug effects* / Q000196:embryology / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D006657:Histones; D018809:Proliferating Cell Nuclear Antigen; D014404:Tubulin; D016159:Tumor Suppressor Protein p53; D006710:Homocysteine
keywords: Chick embryos; Developmental toxicity; Hyperhomocysteinemia; Neural tube defects; Neuronal differentiation; Proliferation