p21WAF1 expression during spermatogenesis of the normal and X-irradiated rat.

The cyclin dependent kinase inhibitor p21WAF1 has been shown to be upregulated during differentiation and after DNA damage in somatic cells. We examined the expression of p21WAF1 mRNA during the differentiation of germ cells in normal and X-irradiated rat testis by in situ hybridization and Northern blotting. p21WAF1 was normally expressed in primary spermatocytes of the pachytene phase, but could also be detected in round spermatids. In preparations of defined segments of the seminiferous tubules, the strongest hybridization signals were detected in the segments containing stages VII VIII and IX XII of the seminiferous epithelium. Ionizing radiation (1-12 Gy) induced the expression of p21WAF1 in a dose-dependent manner and the lowest dose that showed a clear increase in mRNA levels was 3 Gy. The p21WAF1 mRNA levels peaked after 3-4 hours, but remained high compared with the control levels during the 24-h follow-up. No change in the in situ hybridization pattern was seen when comparing unirradiated and irradiated tissue. Thus, it appears that X-irradiation induces p21WAF1 in the pachytene spermatocytes. Since p21WAF1 mRNA was found in pachytene spermatocytes and in round spermatids in normal testis, the protein may take part in the regulation of meiosis and in the 'terminal' differentiation of the male germ cells.