Periodic Reporting for period 1 - DevCycle (Timing cell cycles in multicellular development)
Período documentado: 2022-06-01 hasta 2024-11-30
The development of animals depends on the precise timing of cell divisions, which is crucial for creating tissues and organs with specific structures and functions. As cells differentiate and progress through development, they must adjust the expression of cell-cycle regulators to produce tissue-specific division patterns. However, how cells change their cell-cycle gene-expression programs during development, and how these changes are timed during development, remains largely unexplored. This project aims to investigate these questions by examining intestinal cell cycles in nematodes. In our group, we have recently developed techniques to measure changes in mRNA expression and chromatin modifications in purified intestinal cells from C. elegans. These methods are helping us uncover how cell-cycle gene expression changes during development and identify new regulators that govern stage-specific gene expression. Additionally, we will use an innovative microfluidics platform for long-term imaging of larval development to explore the mechanisms that allow temporal coupling of cell cycle and development. This system will enable us to visualize and manipulate transcription factor gradients that regulate intestinal cell cycles, providing unprecedented insights into the timing of cell cycles. Lastly, I will investigate cell-cycle mechanisms in P. redivivus, a related nematode with a slightly different cell-cycle pattern in its intestinal cells. Studying this species will enhance our understanding of how cell-cycle patterns can vary to produce phenotypic diversity. Overall, this research program aims to provide a mechanistic understanding of how cell cycles are temporally controlled during development, uncovering new insights into the origins of tissue-specific division patterns. This knowledge could pave the way for future applications in tissue engineering and the control of cell divisions in disease contexts.
We have optimized conditions to generate RNA-expression and chromatin profiles from purified intestinal cells from C. elegans, which we are now using to understand how the expression of cell-cycle genes is changing during development. We have also generated AID knock-ins on candidate regulators of cell-cycle transitions, which allows us to deplete endogenous proteins during specific cell-cycle moments and study whether these proteins are important for cell-cycle progression. Finally, we have set up the new microfluidics platform and optimized imaging conditions that now allow us to follow cell-cycle transitions throughout multiple larval stages.
By combining tissue-specific RNA and chromatin profiling with live-imaging of cell-cycle dynamics we are gaining unprecedented resolution on how intestinal cell cycles are changing during C. elegans development. We expect that by the end of the project, we will have identified regulators and mechanisms by which key cells-cycle transitions occur. Despite that our focus has been on the C. elegans intestinal lineage, we think our findings are broadly relevant to many different cell types, and will help uncover how cell cycles are modified during cell-differentiation and tissue formation.