title: Differences in DNA double strand breaks repair in male germ cell types: lessons learned from a differential expression of Mdc1 and 53BP1.
abstract: In male germ cells the repair of DNA double strand breaks (DSBs) differs from that described for somatic cell lines. Irradiation induced immunofluorescent foci (IRIF's) signifying a double strand DNA breaks, were followed in spermatogenic cells up to 16 h after the insult. Foci were characterised for Mdc1, 53BP1 and Rad51 that always were expressed in conjecture with gamma-H2AX. Subsequent spermatogenic cell types were found to have different repair proteins. In early germ cells up to the start of meiotic prophase, i.e. in spermatogonia and preleptotene spermatocytes, 53BP1 and Rad51 are available but no Mdc1 is expressed in these cells before and after irradiation. The latter might explain the radiosensitivity of spermatogonia. Spermatocytes from shortly after premeiotic S-phase till pachytene in epithelial stage IV/V express Mdc1 and Rad51 but no 53BP1 which has no role in recombination involved repair during the early meiotic prophase. Mdc1 is required during this period as in Mdc1 deficient mice all spermatocytes enter apoptosis in epithelial stage IV when they should start mid-pachytene phase of the meiotic prophase. From stage IV mid pachytene spermatocytes to round spermatids, Mdc1 and 53BP1 are expressed while Rad51 is no longer expressed in the haploid round spermatids. Quantifying foci numbers of gamma-H2AX, Mdc1 and 53BP1 at various time points after irradiation revealed a 70% reduction after 16 h in pachytene and diplotene spermatocytes and round spermatids. Although the DSB repair efficiency is higher then in spermatogonia where only a 40% reduction was found, it still does not compare to somatic cell lines where a 70% reduction occurs in 2 h. Taken together, DNA DSBs repair proteins differ for the various types of spermatogenic cells, no germ cell type possessing the complete set. This likely leads to a compromised efficiency relative to somatic cell lines. From the evolutionary point of view it may be an advantage when germ cells die from DNA damage rather than risk the acquisition of transmittable errors made during the repair process.
authors: Department of Endocrinology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. E.A.A.AhmedAbdelGaber@bio.uu.nl
authors: Emad A
authors: EA
authors: Ahmed
authors: Aniek
authors: A
authors: van der Vaart
authors: Angeliqué
authors: A
authors: Barten
authors: Henk B
authors: HB
authors: Kal
authors: Junjie
authors: J
authors: Chen
authors: Zhenkun
authors: Z
authors: Lou
authors: Katherine
authors: K
authors: Minter-Dykhouse
authors: Jirina
authors: J
authors: Bartkova
authors: Jiri
authors: J
authors: Bartek
authors: Peter
authors: P
authors: de Boer
authors: Dirk G
authors: DG
authors: de Rooij
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004247:DNA / Q000528:radiation effects*; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004268:DNA-Binding Proteins; D004307:Dose-Response Relationship, Radiation; D005455:Fluorescent Antibody Technique; D005854:Germ Cells / Q000378:metabolism / Q000528:radiation effects; D006657:Histones / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D008297:Male; D008540:Meiosis / Q000502:physiology; D051379:Mice; D008815:Mice, Inbred Strains; D018345:Mice, Knockout; D010750:Phosphoproteins / Q000378:metabolism*; D051135:Rad51 Recombinase / Q000378:metabolism; D013090:Spermatocytes / Q000528:radiation effects*; D013093:Spermatogonia / Q000378:metabolism; D000071857:Tumor Suppressor p53-Binding Protein 1; D014965:X-Rays
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; C508937:MDC1 protein, mouse; D010750:Phosphoproteins; C498302:Trp53bp1 protein, mouse; D000071857:Tumor Suppressor p53-Binding Protein 1; C475961:gamma-H2AX protein, mouse; D004247:DNA; D051135:Rad51 Recombinase