Powell-Coffman J A, Knight J, Wood W B
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder 80309-0347, USA.
Dev Biol. 1996 Sep 15;178(2):472-83. doi: 10.1006/dbio.1996.0232.
Cleavage and gastrulation initiation in Caenorhabditis elegans embryos are characterized by an invariant temporal and spatial pattern of cell divisions and cell movements. Although bulk embryonic transcription does not begin until gastrulation onset, some transcription can be detected as early as the 4-cell stage. To determine whether any early transcripts are required for normal cleavage-stage patterning, we blocked transcription in embryos by injecting hermaphrodite parental gonads with RNA antisense to the ama-1 gene, which encodes the large subunit of RNA polymerase II. This treatment prevented the expression of a reporter gene driven by an early embryonic promoter but did not detectably perturb the maternally controlled segregation of the germ line P granules or the pattern of cell division through the first four cleavages. In the fifth cell cycle, however, the two endodermal precursor (E) cells divided early and abnormally and failed to initiate gastrulation. The embryos arrested between the sixth and seventh cell cycles with less than 100 cells. These results indicate that embryonically transcribed gene products are required for gastrulation initiation. They also demonstrate the efficacy of a method for blocking embryonic transcription that may be useful in other organisms.
秀丽隐杆线虫胚胎的卵裂和原肠胚形成起始的特征是细胞分裂和细胞运动具有不变的时间和空间模式。虽然整体胚胎转录直到原肠胚形成开始才启动,但早在4细胞阶段就能检测到一些转录。为了确定正常卵裂期模式形成是否需要任何早期转录本,我们通过向雌雄同体亲代性腺注射与ama-1基因反义的RNA来阻断胚胎中的转录,ama-1基因编码RNA聚合酶II的大亚基。这种处理阻止了由早期胚胎启动子驱动的报告基因的表达,但未明显干扰母系控制的生殖系P颗粒的分离或前四次卵裂的细胞分裂模式。然而,在第五个细胞周期中,两个内胚层前体细胞(E)过早且异常地分裂,并且未能启动原肠胚形成。胚胎在第六和第七个细胞周期之间停滞,细胞数量少于100个。这些结果表明,胚胎转录的基因产物是原肠胚形成起始所必需的。它们还证明了一种阻断胚胎转录方法的有效性,该方法可能对其他生物体有用。