Geremia R, Boitani C, Conti M, Monesi V
Cell Differ. 1977 Mar;5(5-6):343-55. doi: 10.1016/0045-6039(77)90072-0.
The rate of RNA synthesis at different stages of spermatogenesis in the mouse, and the preservation of RNA from the diploid to the haploid phase of spermatogenesis were studied in homogeneous germ cell fractions separated by velocity sedimentation at unit gravity. The uridine pool expansion method was used for determining the rate of RNA synthesis: seminiferous tubules were labelled in culture with increasing concentrations of [3H]-uridine and the incorporated radioactivity was estimated in cell fractions separated by velocity sedimentation. The results indicate that before nuclear elongation, round spermatids (steps 1 to 8 of spermiogenesis) synthesize RNA at the same rate per DNA content as middle-late pachytene spermatocytes. The preservation of RNA molecules synthesized in meiosis was investigated by labelling pachytene spermatocytes with T3H]uridine in vivo and collecting samples of germ cells at definite stages of spermatogenesis at various time intervals thereafter. The results show that a considerable proportion the RNA synthesized during the pachytene stage is preserved through spermatid development until late spermiogenesis.
利用单位重力下的速度沉降法分离出的纯一性生殖细胞组分,研究了小鼠精子发生不同阶段的RNA合成速率,以及从精子发生的二倍体阶段到单倍体阶段RNA的保留情况。采用尿苷库扩增法测定RNA合成速率:在培养过程中,用浓度递增的[3H] - 尿苷标记生精小管,并在通过速度沉降分离的细胞组分中估算掺入的放射性。结果表明,在核伸长之前,圆形精子细胞(精子形成的第1至8步)按每个DNA含量计算,其RNA合成速率与中晚期粗线期精母细胞相同。通过在体内用[3H] - 尿苷标记粗线期精母细胞,并在之后的不同时间间隔收集精子发生特定阶段的生殖细胞样本,研究了减数分裂过程中合成的RNA分子的保留情况。结果显示,在粗线期合成的相当一部分RNA在精子细胞发育过程中得以保留,直至精子形成后期。