首页> 外文期刊>Nucleic Acids Research >Mixed spermatogenic germ cell nuclear extracts exhibit high base excisionrepair activity
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Mixed spermatogenic germ cell nuclear extracts exhibit high base excisionrepair activity

机译:混合的生精生殖细胞核提取物具有高碱基切除修复活性

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摘要

Spermatogenic cells exhibit a lower spontaneous mutation frequency than somatic tissues in a lad transgene and many base excision repair (BER) genes display the highest observed level of expression-in the:testis. In this study, uracil-DNA glycosylase-initiated BER activity was measured in nuclear extracts prepared from tissues obtained from each of three mouse strains, Extracts from mixed spermatogenic germ cells displayed the greatest activity followed by liver then brain for all three strains, and the activity for a given tissue was consistent among the three strains, Levels of various BER proteins were examined by western blot analyses and-found to be consistent with activity levels. Nuclear extracts prepared from purified Sertoli cells, a somatic component of the seminiferous epithelium, exhibited significantly lower activity than mixed spermatogenic cell-type nuclear extracts, thereby suggesting that the high BER activity observed in mixed germ cell nuclear extracts was not a characteristic of all testicular cell types, Nuclear extracts from: thymocytes and small intestines were assayed to assess activity in a mitotically active cell type: and tissue. Overall, the order of tissues/cells exhibiting the greatest to lowest activity was mixed germ cells > Sertoli cells > thymocytes > small intestine > liver > brain.
机译:生精细胞在拉德转基因中显示出比体细胞组织更低的自发突变频率,许多碱基切除修复(BER)基因在睾丸中观察到最高的表达水平。在这项研究中,在从三种小鼠品系中的每一种获得的组织中制备的核提取物中,测量了尿嘧啶-DNA糖基化酶引发的BER活性,从混合的生精生殖细胞中提取的品显示出最大的活性,其次是所有三种品系中的肝脏和大脑。在三种菌株中,给定组织的活性是一致的。通过蛋白质印迹分析检查了各种BER蛋白的水平,发现与活性水平一致。由纯化的Sertoli细胞(生精上皮的体细胞成分)制备的核提取物,其活性明显低于混合的生精细胞型核提取物,因此表明,在混合生殖细胞核提取物中观察到的高BER活性并非所有睾丸的特征。检测来自胸腺细胞和小肠的核提取物,以评估在有丝分裂活性细胞类型和组织中的活性。总体而言,表现出最大至最低活性的组织/细胞的顺序是混合生殖细胞>睾丸支持细胞>胸腺细胞>小肠>肝脏>脑。

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