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Nucleotide Excision Repair Activity Varies Among Murine Spermatogenic Cell Types¹

Department of Cellular & Structural Biology,³ Department of Pathology,⁴ Barshop Institute for Longevity and Aging Studies,⁵ San Antonio Cancer Institute,⁶ The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229-3900 Department of Biological Sciences,⁷ Stanford University, Stanford, California 94305-5020 Department of Carcinogenesis,⁸ The University of Texas M.D. Anderson Cancer Center, Science Park/Research Division, Smithville, Texas 78957 Radioisotope Center,⁹ Nara Medical University, Kashihara, Nara 634-0821, Japan Department of Biology,¹⁰ University of Texas at San Antonio, San Antonio, Texas 78249 Department of Chemistry and Biochemistry,¹¹ Southwest Texas State University, San Marcos, Texas 78666 South Texas Veteran's Health Care System,¹² San Antonio, Texas 78201

Germ cells perform a unique and critical biological function: they propagate the DNA that will be used to direct development of the next generation. Genetic integrity of germ cell DNA is essential for producing healthy and reproductively fit offspring, and yet germ cell DNA is damaged by endogenous and exogenous agents. Nucleotide excision repair (NER) is an important mechanism for coping with a variety of DNA lesions. Little is known about NER activity in spermatogenic cells. We expected that germ cells would be more efficient at DNA repair than somatic cells, and that this efficiency may be reduced with age when the prevalence of spontaneous mutations increases. In the present study, NER was measured in defined spermatogenic cell types, including premeiotic cells (A and B type spermatogonia), meiotic cells (pachytene spermatocytes), and postmeiotic haploid cells (round spermatids) and compared with NER in keratinocytes. Global genome repair and transcription-coupled repair subpathways of NER were examined. All spermatogenic cell types from young mice displayed good repair of (6-4) pyrimidone photoproducts, although the repair rate was slower than in primary keratinocytes. In aged mice, repair of 6-4 pyrimidone photoproducts was depressed in postmeiotic cells. While repair of cyclobutane pyrimidine dimers was not detected in spermatogenic cells or in keratinocytes, the transcribed strands of active genes were repaired with greater efficiency than nontranscribed strands or inactive genes in keratinocytes and in meiotic and postmeiotic cells; spermatogonia displayed low to moderate ability to repair cyclobutane pyrimidine dimers on both DNA strands regardless of transcriptional status. Overall, the data suggest cell type-specific NER activity during murine spermatogenesis, and our results have possible implications for germ cell aging.

aging, gamete biology, gametogenesis, spermatogenesis

² Correspondence: Christi Walter, Department of Cellular & Structural Biology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900. FAX: 210 567 3803; walter{at}uthscsa.edu

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