Hamilton Thais Rose dos Santos, de Castro Letícia Signori, Delgado Juliana de Carvalho, de Assis Patrícia Monken, Siqueira Adriano Felipe Perez, Mendes Camilla Mota, Goissis Marcelo Demarchi, Muiño-Blanco Teresa, Cebrián-Pérez José Álvaro, Nichi Marcílio, Visintin José Antonio, D'Ávila Assumpção Mayra Elena Ortiz
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.
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
Reproduction. 2016 Apr;151(4):379-90. doi: 10.1530/REP-15-0403. Epub 2016 Jan 25.
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.
活性氧的作用、鱼精蛋白缺陷和细胞凋亡被认为是精子DNA碎片化最重要的病因。本研究评估了诱导脂质过氧化易感性对天然精液特征的影响,并确定了圣伊内斯公羊精子样本中精子DNA碎片化和睾丸抗氧化防御所涉及的机制。在9周的时间里,每周从12只成年公羊(Ovis aries)采集精液。进行了精子分析(活力、群体活力、异常情况、膜和顶体状态、线粒体电位、DNA碎片化、脂质过氧化和细胞内自由基产生);鱼精蛋白缺乏;PRM1、TNP1和TNP2基因表达;以及谷胱甘肽过氧化物酶(GPx)、谷胱甘肽还原酶、过氧化氢酶(CAT)和超氧化物歧化酶活性的测定以及精浆中的免疫检测。根据用抗坏血酸和硫酸亚铁诱导后精子对脂质过氧化的易感性,将样本分为四组(低、中、高和非常高)。结果通过GLM检验和事后最小显著差异进行分析。我们观察到对诱导脂质过氧化易感性高的组中,天然GPx活性和CAT免疫检测增加。我们还发现对诱导脂质过氧化易感性最高的组中,总精子缺陷、顶体和膜损伤增加。此外,对脂质过氧化易感性较高的组中,线粒体膜电位低、易发生染色质碎片化以及PRM1 mRNA增加。公羊精子对脂质过氧化的易感性可能会损害精子质量,并通过氧化应激干扰氧化稳态,这可能是公羊精子染色质损伤的主要原因。