Nucleic Acids Research, 2001, Vol. 29, No. 6 1366-1372
© 2001 Oxford University Press
Mixed spermatogenic germ cell nuclear extracts exhibit high base excision repair activity
1Department of Cellular and Structural Biology and 2Department of Pathology, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78284, USA, 3Department of Chemistry and Biochemistry, Southwest Texas State University, San Marcos, TX 78666, USA, 4Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, TX 78245, USA and 5South Texas Veterans Health Care System, Audie L. Murphy Hospital, San Antonio, TX 78284, USA
Spermatogenic cells exhibit a lower spontaneous mutation frequency than somatic tissues in a lacI 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.
* To whom correspondence should be addressed at: Department of Cellular and Structural Biology, Mail Code 7762, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78284-3900, USA. Tel: +1 210 567 3832; Fax: +1 210 567 3803; Email: walter{at}uthscsa.edu
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