In vitro approaches to study centriole and cilium function in early mouse embryogenesis

利用体外方法研究小鼠早期胚胎发育过程中中心粒和纤毛的功能

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作者:Isabella Voelkl ,Tamara Civetta ,Mirijam Egg ,Marie Huber ,Songjie Feng ,Alexander Dammermann ,Christa Buecker

Abstract

Although centrioles and primary cilia play an essential role in early mammalian development, their specific function during the interval between their initial formation and the subsequent arrest of embryogenesis in embryos deficient in centrioles or cilia remains largely unexplored. Here, we demonstrate that different 3D in vitro model systems recapitulate early centriole and cilium formation in mouse development. Centrioles and cilia are dispensable in 3D in vitro mouse rosettes, a model system that mimics key events of implantation, including polarization and lumenogenesis. In gastruloids, a model system that recapitulates developmental processes up to E8.5, centriole loss results in early disassembly that can be rescued by additional p53 deletion. In contrast, cells devoid of cilia continue to form elongated, differentiated and polarized gastruloids, with minor differences at 96 h. Finally, we show that in a mutant affecting the centriolar distal appendages, cilia are absent from 2D cultures, but are capable of forming in 3D rosettes and gastruloids, highlighting the importance of multifactorial 3D environment setups in developmental studies.

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