Geremia R, Goldberg R B, Bruce W R
Andrologia. 1976;8(2):147-56. doi: 10.1111/j.1439-0272.1976.tb02124.x.
The separation of mouse spermatogenic cell nuclei by sedimentation velocity at unit gravity has been used to determine the timing of histone and "mouse protamine" synthesis, and the turnover of basic nuclear proteins throughout spermatogenesis. Animals were injected with 3H-arginine or 3H-lysine and at various time intervals (2 hours post-label or from 1 to 30 days post-label) germinal cell nuclei preparations were separated on the staput. Labelled histones and mouse protamine were extracted from staput separated nuclei with hydrocholoric acid and fractionated by polyacrylamide gel electrophoresis. Results indicate that histones are synthesized in association with DNA replication in spermatogonia and preleptotene spermatocytes, in pachytene primary spermatocytes and in spermatids stages 11-16, simultaneously with "mouse protamine". Experiments are reported showing that histones synthesized in pachytene primary spermatocytes and in spermatids stages 11-16 are retained in epididymal spermatozoa, while histones synthesized before meiosis are no longer detectable onto chromatin after meiosis.
利用单位重力沉降速度分离小鼠生精细胞核,以确定组蛋白和“小鼠鱼精蛋白”合成的时间,以及整个精子发生过程中碱性核蛋白的周转情况。给动物注射³H-精氨酸或³H-赖氨酸,并在不同时间间隔(标记后2小时或标记后1至30天)在斯塔普特装置上分离生殖细胞核制剂。用盐酸从斯塔普特分离的细胞核中提取标记的组蛋白和小鼠鱼精蛋白,并通过聚丙烯酰胺凝胶电泳进行分级分离。结果表明,组蛋白在精原细胞和前细线期精母细胞、粗线期初级精母细胞以及11 - 16期精子细胞中与DNA复制同时合成,与“小鼠鱼精蛋白”同步。报告的实验表明,在粗线期初级精母细胞和11 - 16期精子细胞中合成的组蛋白保留在附睾精子中,而减数分裂前合成的组蛋白在减数分裂后在染色质上不再可检测到。