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BOR - Papers in Press, published online ahead of print October 23, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.008672
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BIOLOGY OF REPRODUCTION 68, 628–634 (2003)
DOI: 10.1095/biolreprod.102.008672
© 2003 by the Society for the Study of Reproduction, Inc.

Testis

DNA Double-Strand Breaks and {gamma}-H2AX Signaling in the Testis1

Geert Hamer2,a,b, Hermien L. Roepers-Gajadiena,b, Annemarie van Duyn-Goedhartd, Iris S. Gademanc, Henk B. Kalc, Paul P.W. van Buuld, and Dirk G. de Rooija,b

a Department of Endocrinology, Faculty of Biology, Utrecht University, 3584 CH Utrecht, The Netherlands b Departments of Cell Biology c Radiotherapy, UMCU, 3584 CX Utrecht, The Netherlands d MCG-Department of Radiation Genetics and Chemical Mutagenesis, Sylvius Laboratory, Leiden University, 2333 AL Leiden, The Netherlands

Within minutes of the induction of DNA double-strand breaks in somatic cells, histone H2AX becomes phosphorylated at serine 139 and forms {gamma}-H2AX foci at the sites of damage. These foci then play a role in recruiting DNA repair and damage-response factors and changing chromatin structure to accurately repair the damaged DNA. These {gamma}-H2AX foci appear in response to irradiation and genotoxic stress and during V(D)J recombination and meiotic recombination. Independent of irradiation, {gamma}-H2AX occurs in all intermediate and B spermatogonia and in preleptotene to zygotene spermatocytes. Type A spermatogonia and round spermatids do not exhibit {gamma}-H2AX foci but show homogeneous nuclear {gamma}-H2AX staining, whereas in pachytene spermatocytes {gamma}-H2AX is only present in the sex vesicle. In response to ionizing radiation, {gamma}-H2AX foci are generated in spermatogonia, spermatocytes, and round spermatids. In irradiated spermatogonia, {gamma}-H2AX interacts with p53, which induces spermatogonial apoptosis. These events are independent of the DNA-dependent protein kinase (DNA-PK). Irradiation-independent nuclear {gamma}-H2AX staining in leptotene spermatocytes demonstrates a function for {gamma}-H2AX during meiosis. {gamma}-H2AX staining in intermediate and B spermatogonia, preleptotene spermatocytes, and sex vesicles and round spermatids, however, indicates that the function of H2AX phosphorylation during spermatogenesis is not restricted to the formation of {gamma}-H2AX foci at DNA double-strand breaks.

1 This work was supported by the J.A. Cohen Institute for Radiopathology and Radiation Protection, Leiden, The Netherlands.

2 Correspondence: Geert Hamer, Department of Endocrinology, Faculty of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. FAX: 31 30 2532837; g.hamer{at}bio.uu.nl




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