Riesenberg Amy N, Conley Kevin W, Le Tien T, Brown Nadean L
Division of Developmental Biology, Cincinnati Children's Hospital Research Foundation, Cincinnati, Ohio.
Department of Cell Biology & Human Anatomy, University of California Davis, Davis, California.
Dev Dyn. 2018 Jan;247(1):212-221. doi: 10.1002/dvdy.24542. Epub 2017 Aug 2.
Notch signaling is broadly required during embryogenesis, frequently activating the transcription of two basic helix-loop-helix transcription factors, Hes1 and Hes5. But, it remains unresolved when and where Hes1 and Hes5 act alone or together during development. Here, we analyzed a Hes5-green fluorescent protein (GFP) bacterial artificial chromosome (BAC) transgenic mouse, as a proxy for endogenous Hes5. We directly compared transgenic GFP expression with Hes1, and particular markers of embryonic lens and retina development.
Hes5-GFP is dynamic within subsets of retinal and lens progenitor cells, and differentiating retinal ganglion neurons, in contrast to Hes1 found in all progenitor cells. In the adult retina, only Müller glia express Hes5-GFP. Finally, Hes5-GFP is up-regulated in Hes1 germline mutants, consistent with previous demonstration that Hes1 suppresses Hes5 transcription.
Hes5-GFP BAC transgenic mice are useful for identifying Hes5-expressing cells. Although Hes5-GFP and Hes1 are coexpressed in particular developmental contexts, we also noted cohorts of lens or retinal cells expressing just one factor. The dynamic Hes5-GFP expression pattern, coupled with its derepressed expression in Hes1 mutants, suggests that this transgene contains the relevant cis-regulatory elements that regulate endogenous Hes5 in the mouse lens and retina. Developmental Dynamics 247:212-221, 2018. © 2017 Wiley Periodicals, Inc.
Notch信号在胚胎发育过程中广泛需要,经常激活两种基本螺旋-环-螺旋转录因子Hes1和Hes5的转录。但是,在发育过程中Hes1和Hes5何时何地单独或共同发挥作用仍未得到解决。在这里,我们分析了一种Hes5-绿色荧光蛋白(GFP)细菌人工染色体(BAC)转基因小鼠,作为内源性Hes5的替代物。我们直接将转基因GFP表达与Hes1以及胚胎晶状体和视网膜发育的特定标志物进行了比较。
与在所有祖细胞中发现的Hes1不同,Hes5-GFP在视网膜和晶状体祖细胞亚群以及分化的视网膜神经节神经元中是动态的。在成年视网膜中,只有穆勒胶质细胞表达Hes5-GFP。最后,Hes5-GFP在Hes1种系突变体中上调,这与之前证明的Hes1抑制Hes5转录一致。
Hes5-GFP BAC转基因小鼠可用于鉴定表达Hes5的细胞。虽然Hes5-GFP和Hes1在特定的发育背景下共表达,但我们也注意到仅表达一种因子的晶状体或视网膜细胞群。动态的Hes5-GFP表达模式,加上其在Hes1突变体中的去抑制表达,表明该转基因包含在小鼠晶状体和视网膜中调节内源性Hes5的相关顺式调控元件。《发育动力学》247:212 - 221,2018年。©2017威利期刊公司。