Souza Elisson Terêncio, Silva Cláudio Vieira, Travençolo Bruno Augusto Nassif, Alves Benner Geraldo, Beletti Marcelo Emílio
Laboratory of Biology of Reproduction, Institute of Biomedical Sciences, Federal University of Uberlândia, CEP 38400-902, Uberlândia, MG, Brazil.
Faculty of Computer Science, Federal University of Uberlândia, CEP 38400-902, Uberlândia, MG, Brazil.
Reprod Biol. 2018 Jun;18(2):177-181. doi: 10.1016/j.repbio.2018.04.001. Epub 2018 Apr 26.
Alterations in sperm chromatin have been related with subfertility in several mammals. In this study, chromatin alteration types (Base, Basal half, Central axis, Dispersed, and Whole) were assessed by toluidine blue (TB) staining, 6-diamidino-2-fenilindole (DAPI) and anti-protamine 1 antibody (anti-PR1) labeling in sperm samples of fertile and subfertile bulls. Semen samples were obtained from bulls kept in Artificial Insemination Center (fertile bulls) or from bulls subjected to scrotal insulation (subfertile bulls). The percentage of chromatin alterations identified by TB was similar (P > 0.05) in semen samples of fertile and subfertile bulls. In contrast, a greater (P < 0.01) chromatin decondensation and heterogeneity were recorded in semen samples of subfertile bulls. In DAPI and anti-PR1 methods, the subfertile bulls samples had a higher (P < 0.05) percentage of alteration in the base as well as overall chromatin alterations (P < 0.05). Moreover, the chromatin alterations recorded with TB, DAPI, and anti-PR1 were compared in semen samples of fertile and subfertile bulls. In fertile bulls, the overall chromatin alterations were similar (P > 0.05) among the methods In contrast, semen samples of subfertile bulls had a higher (P < 0.05) percentage of overall chromatin alterations when labeled with DAPI. In conclusion, our findings shown that all dye tested had specific sperm stainability and can be feasible to monitor subfertility condition in bulls. Also, different chromatin alteration types in sperm samples of fertile and suberftile bulls were recorded.
精子染色质的改变与多种哺乳动物的生育力低下有关。在本研究中,通过甲苯胺蓝(TB)染色、6-二脒基-2-苯基吲哚(DAPI)和抗鱼精蛋白1抗体(抗-PR1)标记,对生育力正常和低下的公牛精子样本中的染色质改变类型(碱基、基部一半、中心轴、分散和整体)进行了评估。精液样本取自人工授精中心的公牛(生育力正常的公牛)或接受阴囊隔热处理的公牛(生育力低下的公牛)。TB鉴定出的染色质改变百分比在生育力正常和低下的公牛精液样本中相似(P>0.05)。相比之下,生育力低下的公牛精液样本中记录到更大程度(P<0.01)的染色质解聚和异质性。在DAPI和抗-PR1方法中,生育力低下的公牛样本在碱基改变以及整体染色质改变方面的百分比更高(P<0.05)。此外,还比较了生育力正常和低下的公牛精液样本中用TB、DAPI和抗-PR1记录的染色质改变。在生育力正常的公牛中,各方法之间的整体染色质改变相似(P>0.05)。相比之下,用DAPI标记时,生育力低下的公牛精液样本中整体染色质改变的百分比更高(P<0.05)。总之,我们的研究结果表明,所有测试染料都具有特定的精子染色性,并且可用于监测公牛的生育力低下状况。此外,还记录了生育力正常和低下的公牛精子样本中不同的染色质改变类型。