Ainsworth Julia, Zhao Yunchong, Gao Ke, Gravina Nicholas M, Goldberg Zachary H, Pinkerton Coleman, Rifkin Scott A, Ernst Andreas M, Garcia Hernan G, Perry Michael W
Department of Cell and Developmental Biology, School of Biological Sciences, University of California, San Diego, La Jolla, CA 92093, USA.
Department of Physics, University of California, Berkeley, Berkeley, CA 94720, USA.
Curr Biol. 2025 Jun 23;35(12):2946-2959.e5. doi: 10.1016/j.cub.2025.05.037. Epub 2025 Jun 10.
Although transcription happens in bursts, it is unclear whether variation in the rate and pattern of bursting matters during animal development. We examined whether the amount and timing of transcription influence the ratio of cell types produced during stochastic patterning of the Drosophila retina. This system is balanced between 2 outcomes: ∼70% of R7 photoreceptors (PRs) express Rhodopsin 4 (Rh4), and the rest express Rhodopsin 3 (Rh3). Cell fate depends on the cell-intrinsic binary decision to express the transcription factor spineless (Ss). Changing the ss bursting pattern by replacing the ss core promoter led to a different ratio of R7 PR types. We hypothesized that random variation in the timing of transcriptional initiation followed by autoregulation might control the outcome. Instead, we found that the decision occurs before R7 specification and before protein is made, with no feedback via Ss protein. Surprisingly, this happens in a field of progenitor cells that give rise to all retina cell types, which all initially transcribe ss. A subset stops transcribing ss over time. Those that become R7s and maintain ss transcription take the Ss-ON/Rh4 fate. Live imaging of ss transcription suggests increased time spent in off periods could decrease the probability of new transcription and therefore the Ss-ON ratio. Transiently increasing continuity of transcription produces all Ss-ON outcomes, while introducing longer gaps lowers the ratio. Targeting CBP to the ss locus increases the ratio, indicating a repressive role for chromatin state during periods when transcription is inactive. We conclude the ratio is set by the amount and dynamics of ss transcription.
尽管转录是以突发脉冲的形式发生,但尚不清楚在动物发育过程中,突发脉冲的速率和模式变化是否重要。我们研究了转录的量和时间是否会影响果蝇视网膜随机模式形成过程中产生的细胞类型比例。该系统在两种结果之间保持平衡:约70%的R7光感受器(PR)表达视紫红质4(Rh4),其余的表达视紫红质3(Rh3)。细胞命运取决于表达转录因子无脊椎(Ss)的细胞内在二元决定。通过替换ss核心启动子来改变ss突发模式,会导致不同比例的R7 PR类型。我们假设转录起始时间的随机变化随后的自动调节可能控制结果。相反,我们发现该决定发生在R7特化之前和蛋白质产生之前,且不存在通过Ss蛋白的反馈。令人惊讶的是,这发生在一个能产生所有视网膜细胞类型的祖细胞区域,所有这些祖细胞最初都转录ss。随着时间的推移,一部分祖细胞停止转录ss。那些成为R7并维持ss转录的祖细胞采取Ss-ON/Rh4命运。对ss转录的实时成像表明,在关闭期花费的时间增加可能会降低新转录的概率,从而降低Ss-ON比例。短暂增加转录的连续性会产生所有Ss-ON结果,而引入更长的间隔会降低该比例。将CBP靶向到ss基因座会增加该比例,这表明在转录不活跃期间,染色质状态起抑制作用。我们得出结论,该比例是由ss转录的量和动态设定的。