Two new genetically modified mouse alleles labeling distinct phases of retinal ganglion cell development by fluorescent proteins.

Two new genetically modified mouse alleles labeling distinct phases of retinal ganglion cell development by fluorescent proteins.
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两个新的转基因小鼠等位基因通过荧光蛋白标记视网膜神经节细胞发育的不同阶段。

DOI:
10.1002/dvdy.233
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发表时间:
2020-12
期刊:
Developmental dynamics : an official publication of the American Association of Anatomists
影响因子:
--
通讯作者:
Mu X
Mu X
中科院分区:
其他
文献类型:
--
作者:
Ge Y;Wu F;Cheng M;Bard J;Mu X

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在发育过程中,所有类型的视网膜细胞都以逐步的方式从视网膜祖细胞(RPC)中产生。Atoh 7和Pou 4f 2标记视网膜神经节细胞(RGC)发生的两个阶段并在其中发挥作用; Atoh 7在RPC亚群中发挥作用,使其能够胜任RGC命运,而Pou 4f 2参与RGC命运特化和RGC分化。尽管对它们的作用进行了广泛的研究,但这两个因子所代表的两个相位的特性尚未得到很好的研究,这可能是由于视网膜细胞的异质性。在这份报告中,我们描述了两个新的敲入小鼠等位基因,Atoh 7zsGreenCreERT 2和Pou 4f 2FlagtdTomato,它们通过荧光蛋白在RGC发育的两个阶段标记视网膜细胞。此外,Atoh 7zsGreenCreERT 2等位基因允许在他莫昔芬诱导后以剂量依赖性方式间接标记RGC和其他细胞类型。此外,这些等位基因可用于通过荧光辅助细胞分选(FACS)纯化不同阶段的视网膜细胞。对来自Atoh 7zsGreenCreERT 2视网膜的纯化细胞的单细胞RNA-seq分析进一步验证了该等位基因标记了过渡/感受态RPC及其后代(包括RGC)。因此,这两个等位基因是研究RGC形成的分子和遗传机制的非常有用的工具。
During development, all retinal cell types arise from retinal progenitor cells (RPCs) in a step-wise fashion. Atoh7 and Pou4f2 mark, and function in, two phases of retinal ganglion cell (RGC) genesis; Atoh7 functions in a subpopulation of RPCs to render them competent for the RGC fate, whereas Pou4f2 participates in RGC fate specification and RGC differentiation. Despite extensive research on their roles, the properties of the two phases represented by these two factors have not been well studied, likely due to the retinal cellular heterogeneity. In this report, we describe two novel knock-in mouse alleles, Atoh7zsGreenCreERT2 and Pou4f2FlagtdTomato, which labeled retinal cells in the two phases of RGC development by fluorescent proteins. Also, the Atoh7zsGreenCreERT2 allele allowed for indirect labeling of RGCs and other cell types upon tamoxifen induction in a dose-dependent manner. Further, these alleles could be used to purify retinal cells in the different phases by fluorescence assisted cell sorting (FACS). Single cell RNA-seq analysis of purified cells from Atoh7zsGreenCreERT2 retinas further validated that this allele labeled both transitional/competent RPCs and their progenies including RGCs. Thus, these two alleles are very useful tools for studying the molecular and genetic mechanisms underlying RGC formation.
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