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早期小鼠胚胎中转录的可视化。

Visualization of Transcription in early mouse embryos.

作者信息

Hughes M E, Bürki K, Fakan S

出版信息

Chromosoma. 1979 Aug 10;73(2):179-90. doi: 10.1007/BF00331570.

Abstract

Transcriptional activity during early mammalian embryogenesis was examined in 2 cell, 4--8 cell and late morula/early blastocyst mouse embryos. Zona pellucida-free embryos were obtained after pronase digestion followed by a PMSF wash to inhibit proteolytic activity. The embryos were then lysed with NP40 detergent and spread for electron microscopy according to Miller and Bakken (1972). pre-mRNA transcription was observed at all stages. Comparison of growing RNP chain lengths revealed statistically significant differences in the distribution of shorter fibrils between the successive developmental stages examined. The number of shorter fibrils was lower in the 4--8 cell stage than in either of the two other stages. Transcription complexes of ribosomal type were detected only in 4--8 cell and morula/blastocyst embryos. Structures resembling replication loops were observed within chromatin from all stages. Similar loop-like structures as well as individually transcribed RNP fibrils were also occasionally found on fibres emerging from mitotic chromosomes. The results are discussed in the context of recent findings concerning genetic expression in early mouse embryos.

摘要

在2细胞、4-8细胞以及桑葚胚晚期/囊胚早期的小鼠胚胎中检测了早期哺乳动物胚胎发育过程中的转录活性。经胰蛋白酶消化后,用苯甲基磺酰氟洗涤以抑制蛋白水解活性,从而获得无透明带胚胎。然后用NP40去污剂裂解胚胎,并按照米勒和巴肯(1972年)的方法铺展用于电子显微镜观察。在所有阶段均观察到前体mRNA转录。对正在生长的核糖核蛋白链长度的比较显示,在所检查的连续发育阶段之间,较短原纤维的分布存在统计学上的显著差异。4-8细胞阶段较短原纤维的数量低于其他两个阶段中的任何一个。仅在4-8细胞和桑葚胚/囊胚胚胎中检测到核糖体类型的转录复合物。在所有阶段的染色质中均观察到类似复制环的结构。在有丝分裂染色体发出的纤维上偶尔也发现了类似环的结构以及单独转录的核糖核蛋白原纤维。结合最近关于小鼠早期胚胎基因表达的研究结果对这些结果进行了讨论。

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