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

Area of Science:

• Developmental Biology
• Cell Biology
• Biophysics

Background:

• Centrioles and primary cilia are vital for early mammalian development.
• Their precise roles between formation and developmental arrest are unclear.
• 3D in vitro models offer insights into early development.

Purpose of the Study:

• To investigate the function of centrioles and cilia in early mammalian development using 3D in vitro models.
• To explore the impact of centriole and cilium absence on specific developmental processes.
• To assess the influence of the 3D environment on centriole and cilium formation.

Main Methods:

• Utilized 3D in vitro mouse rosettes and gastruloid models.
• Generated centriole- and cilium-deficient cell lines.
• Investigated developmental processes like polarization, lumenogenesis, and gastruloid elongation.
• Analyzed a mutant affecting centriolar distal appendages.

Main Results:

• Centrioles and cilia were dispensable in 3D mouse rosettes during implantation-like events.
• Centriole loss in gastruloids caused early disassembly, rescueable by p53 deletion.
• Cilia-deficient cells formed elongated, polarized gastruloids with minimal 96h differences.
• A distal appendage mutant lacked cilia in 2D but formed them in 3D models.

Conclusions:

• The requirement for centrioles and cilia varies depending on the developmental context and model system.
• 3D environments are crucial for recapitulating and studying complex developmental processes.
• Centriole function is critical for gastruloid integrity, while cilia play a less prominent role in this specific model.