Osborne Nishimura Erin, Zhang Jay C, Werts Adam D, Goldstein Bob, Lieb Jason D
Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, Chapel Hill, North Carolina, United States of America; Department of Biology, University of North Carolina, Chapel Hill, Chapel Hill, North Carolina, United States of America.
Department of Biology, University of North Carolina, Chapel Hill, Chapel Hill, North Carolina, United States of America.
PLoS Genet. 2015 Apr 13;11(4):e1005117. doi: 10.1371/journal.pgen.1005117. eCollection 2015 Apr.
After fertilization but prior to the onset of zygotic transcription, the C. elegans zygote cleaves asymmetrically to create the anterior AB and posterior P1 blastomeres, each of which goes on to generate distinct cell lineages. To understand how patterns of RNA inheritance and abundance arise after this first asymmetric cell division, we pooled hand-dissected AB and P1 blastomeres and performed RNA-seq. Our approach identified over 200 asymmetrically abundant mRNA transcripts. We confirmed symmetric or asymmetric abundance patterns for a subset of these transcripts using smFISH. smFISH also revealed heterogeneous subcellular patterning of the P1-enriched transcripts chs-1 and bpl-1. We screened transcripts enriched in a given blastomere for embryonic defects using RNAi. The gene neg-1 (F32D1.6) encoded an AB-enriched (anterior) transcript and was required for proper morphology of anterior tissues. In addition, analysis of the asymmetric transcripts yielded clues regarding the post-transcriptional mechanisms that control cellular mRNA abundance during asymmetric cell divisions, which are common in developing organisms.
在受精后但合子转录开始之前,秀丽隐杆线虫的合子进行不对称分裂,产生前部的AB和后部的P1卵裂球,每个卵裂球随后都会发育出不同的细胞谱系。为了了解在第一次不对称细胞分裂后RNA遗传和丰度模式是如何产生的,我们将手工解剖的AB和P1卵裂球汇集在一起并进行了RNA测序。我们的方法鉴定出了200多种不对称丰富的mRNA转录本。我们使用单分子荧光原位杂交(smFISH)确认了这些转录本中的一部分的对称或不对称丰度模式。smFISH还揭示了富含P1的转录本chs-1和bpl-1的异质亚细胞模式。我们使用RNA干扰筛选了在特定卵裂球中富集的转录本,以寻找胚胎缺陷。基因neg-1(F32D1.6)编码一种富含AB(前部)的转录本,是前部组织正常形态所必需的。此外,对不对称转录本的分析提供了有关转录后机制的线索,这些机制在不对称细胞分裂过程中控制细胞mRNA丰度,而不对称细胞分裂在发育中的生物体中很常见。