Jakobsen Anne S, Avery Birthe, Dieleman Steph J, Knijn Hiemke M, Vos Peter L A M, Thomsen Preben D
Department of Large Animal Sciences, Veterinary Reproduction and Obstetrics, The Royal Veterinary and Agricultural University, Frederiksberg C, Denmark.
Mol Reprod Dev. 2006 Feb;73(2):196-205. doi: 10.1002/mrd.20389.
Transcription from the embryos own ribosomal genes is initiated in most species at the same time as the maternal-embryonic transition. Recently data have indicated that a minor activation may take place during the third embryonic cell cycle in the bovine, one cell cycle before the major activation of the embryonic genome. In the present study, ribosomal RNA (rRNA) transcription was investigated by visualization of the rRNA by fluorescent in situ hybridization, and subsequent visualization of the argyrophilic nucleolar proteins by silver staining. A total of 145 in vivo developed and 200 in vitro produced bovine embryos were investigated to allow comparison of transcription initiation. Signs of active transcription of rRNA were observed in the third cell cycle in 29% of the in vitro produced embryos (n = 35) and in 58% of the in vivo developed embryos (n = 11). Signs of active transcription of rRNA were not apparent in the early phase of the fourth cell cycle but restarted later on. All embryos in the fifth or later cell cycles were all transcribing rRNA. The signs of rRNA synthesis during the third and fourth embryonic cell cycle could be blocked by actinomycin D, which is a strong inhibitor of RNA polymerase I. In conclusion, rRNA transcription is initiated during the third cell cycle at a low level in both in vivo developed and in vitro produced bovine embryos. Transcription seems to be interrupted during the G1 phase of the fourth cell cycle, but reinitiates in the late half of the cycle and persists during subsequent cell cycles.
在大多数物种中,胚胎自身核糖体基因的转录与母源-胚胎转换同时启动。最近的数据表明,在牛中,可能在第三个胚胎细胞周期发生轻微激活,比胚胎基因组的主要激活早一个细胞周期。在本研究中,通过荧光原位杂交可视化核糖体RNA(rRNA),随后通过银染可视化嗜银核仁蛋白,对rRNA转录进行了研究。共研究了145个体内发育的和200个体外生产的牛胚胎,以比较转录起始情况。在第三个细胞周期中,29%的体外生产胚胎(n = 35)和58%的体内发育胚胎(n = 11)观察到rRNA活跃转录的迹象。在第四个细胞周期的早期阶段,rRNA活跃转录的迹象不明显,但随后重新开始。第五个或更晚细胞周期的所有胚胎都在转录rRNA。第三个和第四个胚胎细胞周期中rRNA合成的迹象可被放线菌素D阻断,放线菌素D是RNA聚合酶I的强抑制剂。总之,在体内发育和体外生产的牛胚胎中,rRNA转录在第三个细胞周期开始时水平较低。转录似乎在第四个细胞周期的G1期中断,但在周期后半段重新启动,并在随后的细胞周期中持续。