title: Induced lipid peroxidation in ram sperm: semen profile, DNA fragmentation and antioxidant status.
abstract: Action of reactive oxygen species, protamination failures and apoptosis are considered the most important etiologies of sperm DNA fragmentation. This study evaluated the effects of induced lipid peroxidation susceptibility on native semen profile and identified the mechanisms involved in sperm DNA fragmentation and testicular antioxidant defense on Santa Ines ram sperm samples. Semen was collected from 12 adult rams (Ovis aries) performed weekly over a 9-week period. Sperm analysis (motility, mass motility, abnormalities, membrane and acrosome status, mitochondrial potential, DNA fragmentation, lipid peroxidation and intracellular free radicals production); protamine deficiency; PRM1, TNP1 and TNP2 gene expression; and determination of glutathione peroxidase (GPx), glutathione reductase, catalase (CAT) and superoxide dismutase activity and immunodetection in seminal plasma were performed. Samples were distributed into four groups according to the sperm susceptibility to lipid peroxidation after induction with ascorbate and ferrous sulfate (low, medium, high and very high). The results were analyzed by GLM test and post hoc least significant difference. We observed an increase in native GPx activity and CAT immunodetection in groups with high susceptibility to induced lipid peroxidation. We also found an increase in total sperm defects, acrosome and membrane damages in the group with the highest susceptibility to induced lipid peroxidation. Additionally, the low mitochondrial membrane potential, susceptible to chromatin fragmentation and the PRM1 mRNA were increased in the group showing higher susceptibility to lipid peroxidation. Ram sperm susceptibility to lipid peroxidation may compromise sperm quality and interfere with the oxidative homeostasis by oxidative stress, which may be the main cause of chromatin damage in ram sperm.
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: Thais Rose dos Santos
authors.initials: TR
authors.lastname: Hamilton
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: Letícia Signori
authors.initials: LS
authors.lastname: de Castro
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: Juliana de Carvalho
authors.initials: Jde C
authors.lastname: Delgado
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: Patrícia Monken
authors.initials: PM
authors.lastname: de Assis
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: Adriano Felipe Perez
authors.initials: AF
authors.lastname: Siqueira
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: Camilla Mota
authors.initials: CM
authors.lastname: Mendes
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: Marcelo Demarchi
authors.initials: MD
authors.lastname: Goissis
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: Teresa
authors.initials: T
authors.lastname: Muiño-Blanco
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: José Álvaro
authors.initials: JÁ
authors.lastname: Cebrián-Pérez
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: Marcílio
authors.initials: M
authors.lastname: Nichi
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain.
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Visintin
authors.affiliation: Department of Animal ReproductionSchool of Veterinary Medicine and Animal Science, University of Sao Paulo. Avenida Prof. Dr. Orlando Marques de Paiva, 87, 05508 270, Cidade Universitária, Sao Paulo, BrazilGrupo de Biología y Fisiología de la ReproducciónFacultad de Veterinaria, Instituto de Investigación de Ciencias Ambientales de Aragón (IUCA), Department of Biochemistry and Molecular and Cell Biology, University of Zaragoza, Miguel Servet, 177, 50013 Zaragoza, Spain meoaa@usp.br.
authors.forename: Mayra Elena Ortiz
authors.initials: ME
authors.lastname: D'Ávila Assumpção
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D017209:Apoptosis; D015153:Blotting, Western; D002478:Cells, Cultured; D053938:DNA Fragmentation; D015227:Lipid Peroxidation; D008297:Male; D053078:Membrane Potential, Mitochondrial; D018384:Oxidative Stress; D012333:RNA, Messenger / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012661:Semen / Q000378:metabolism*; D055101:Semen Analysis / Q000379:methods*; D034561:Sheep, Domestic; D013094:Spermatozoa / Q000737:chemistry / Q000378:metabolism*; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
chemical_list: D000975:Antioxidants; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D017392:Thiobarbituric Acid Reactive Substances