MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord Cells

The neurotoxicity caused by methylmercury (MeHg) is well documented; however, the developmental neurotoxicity in spinal cord is still not fully understood. Here we investigated whether MeHg affects the spinal cord layers development. Chicken embryos at E3 were treated in ovo with 0.1 μg MeHg/50 μL s...

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Main Authors: Fabiana F. Ferreira, Dib Ammar, Gilian F. Bourckhardt, Karoline Kobus-Bianchini, Yara M. R. Müller, Evelise M. Nazari
Format: Article
Language:English
Published: Wiley 2015-01-01
Series:Journal of Toxicology
Online Access:http://dx.doi.org/10.1155/2015/532691
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author Fabiana F. Ferreira
Dib Ammar
Gilian F. Bourckhardt
Karoline Kobus-Bianchini
Yara M. R. Müller
Evelise M. Nazari
author_facet Fabiana F. Ferreira
Dib Ammar
Gilian F. Bourckhardt
Karoline Kobus-Bianchini
Yara M. R. Müller
Evelise M. Nazari
author_sort Fabiana F. Ferreira
collection DOAJ
description The neurotoxicity caused by methylmercury (MeHg) is well documented; however, the developmental neurotoxicity in spinal cord is still not fully understood. Here we investigated whether MeHg affects the spinal cord layers development. Chicken embryos at E3 were treated in ovo with 0.1 μg MeHg/50 μL saline solution and analyzed at E10. Thus, we performed immunostaining using anti-γ-H2A.X to recognize DNA double-strand breaks and antiphosphohistone H3, anti-p21, and anti-cyclin E to identify cells in proliferation and cell cycle proteins. Also, to identify neuronal cells, we used anti-NeuN and anti-βIII-tubulin antibodies. After the MeHg treatment, we observed the increase on γ-H2A.X in response to DNA damage. MeHg caused a decrease in the proliferating cells and in the thickness of spinal cord layers. Moreover, we verified that MeHg induced an increase in the number of p21-positive cells but did not change the cyclin E-positive cells. A significantly high number of TUNEL-positive cells indicating DNA fragmentation were observed in MeHg-treated embryos. Regarding the neuronal differentiation, MeHg induced a decrease in NeuN expression and did not change the expression of βIII-tubulin. These results showed that in ovo MeHg exposure alters spinal cord development by disturbing the cell proliferation and death, also interfering in early neuronal differentiation.
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spelling doaj-art-c77f0eb0a6fe4cc8b7315c6fb8b7a3942025-02-03T01:26:46ZengWileyJournal of Toxicology1687-81911687-82052015-01-01201510.1155/2015/532691532691MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord CellsFabiana F. Ferreira0Dib Ammar1Gilian F. Bourckhardt2Karoline Kobus-Bianchini3Yara M. R. Müller4Evelise M. Nazari5Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, 88040-900 Florianópolis, SC, BrazilDepartamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, 88040-900 Florianópolis, SC, BrazilDepartamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, 88040-900 Florianópolis, SC, BrazilDepartamento de Fisioterapia, Centro de Ciências da Saúde e do Esporte, UDESC, Rua Pascoal Simone 358, Coqueiros, 88080-350 Florianópolis, SC, BrazilDepartamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, 88040-900 Florianópolis, SC, BrazilDepartamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, UFSC, Campus Universitário, Trindade, 88040-900 Florianópolis, SC, BrazilThe neurotoxicity caused by methylmercury (MeHg) is well documented; however, the developmental neurotoxicity in spinal cord is still not fully understood. Here we investigated whether MeHg affects the spinal cord layers development. Chicken embryos at E3 were treated in ovo with 0.1 μg MeHg/50 μL saline solution and analyzed at E10. Thus, we performed immunostaining using anti-γ-H2A.X to recognize DNA double-strand breaks and antiphosphohistone H3, anti-p21, and anti-cyclin E to identify cells in proliferation and cell cycle proteins. Also, to identify neuronal cells, we used anti-NeuN and anti-βIII-tubulin antibodies. After the MeHg treatment, we observed the increase on γ-H2A.X in response to DNA damage. MeHg caused a decrease in the proliferating cells and in the thickness of spinal cord layers. Moreover, we verified that MeHg induced an increase in the number of p21-positive cells but did not change the cyclin E-positive cells. A significantly high number of TUNEL-positive cells indicating DNA fragmentation were observed in MeHg-treated embryos. Regarding the neuronal differentiation, MeHg induced a decrease in NeuN expression and did not change the expression of βIII-tubulin. These results showed that in ovo MeHg exposure alters spinal cord development by disturbing the cell proliferation and death, also interfering in early neuronal differentiation.http://dx.doi.org/10.1155/2015/532691
spellingShingle Fabiana F. Ferreira
Dib Ammar
Gilian F. Bourckhardt
Karoline Kobus-Bianchini
Yara M. R. Müller
Evelise M. Nazari
MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord Cells
Journal of Toxicology
title MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord Cells
title_full MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord Cells
title_fullStr MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord Cells
title_full_unstemmed MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord Cells
title_short MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord Cells
title_sort mehg developing exposure causes dna double strand breaks and elicits cell cycle arrest in spinal cord cells
url http://dx.doi.org/10.1155/2015/532691
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