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Assessment of plasma membrane and chromatin structure of sperm from transgenic and non-transgenic boars

机译:转基因和非转基因公猪精子质膜和染色质结构的评估

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The aim of this study was to determine the apoptotic changes and chromatin damage in non-transgenic and transgenic boars carrying the human l1,2-fucosyltransferase gene. Five ejaculates were collected from six transgenic (TG) and six non-transgenic (NTG) boars. Five ejaculates were collected from six transgenic (TG) and six non-transgenic (NTG) boars both crossbreds of Polish Landrace and Large White. Two fluorescence methods were employed to measure apoptosis: an assay to assess the early changes in sperm membrane integrity using fluorophore YO-PRO-1 and an assay for phosphatidylserine (PS) translocation across the plasma membrane using fluorescein-labeled Annexin-V. The chromatin damage was assessed based on the sperm chromatin structure assay method. No significant differences in the proportion of all detected subpopulations of spermatozoa were found between TG and NTG boars. Similarly, the analysis of the chromatin structure revealed no statistical differences in the sperm chromatin damage between TG and NTG boars. In conclusion, the presence of the human l1,2-fucosyltransferase gene in the genome of TG boars did not cause any spermatogenesis process disturbances leading to the increased production of apoptotic spermatozoa. Moreover, the low level of sperm with damaged chromatin in TG boars confirms the high stability of the spermatogenesis process in the TG boars analyzed.
机译:这项研究的目的是确定携带人类11,2-岩藻糖基转移酶基因的非转基因和转基因公猪的凋亡变化和染色质损伤。从六只转基因(TG)和六只非转基因(NTG)公猪中收集了五个射精。从六只转基因公猪和六只非转基因公猪中收集了五种射精,这两种公猪都是波兰长白猪和大白猪的杂交种。两种荧光方法用于测量细胞凋亡:一种使用荧光团YO-PRO-1评估精子膜完整性早期变化的测定法,另一种使用荧光素标记的Annexin-V进行质膜磷脂酰丝氨酸(PS)转运的测定法。基于精子染色质结构测定方法评估染色质损伤。在TG和NTG公猪之间发现的所有精子亚群的比例没有显着差异。同样,对染色质结构的分析表明,TG和NTG公猪之间的精子染色质损伤无统计学差异。总之,TG公猪基因组中人类11,2-岩藻糖基转移酶基因的存在不会引起任何精子发生过程紊乱,从而导致凋亡性精子产生增加。此外,TG公猪中染色质受损的精子水平低,证实了所分析的TG公猪的生精过程具有很高的稳定性。

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