Cell Cycle Regulation of Morphogenetic Processes during Development

During organismal development, differential regulation of the cell cycle is critical to many cell biological processes, including cell migration, cell fate specification and differentiation. While the mechanisms of cell cycle regulation are well studied, how control of the cell cycle is linked to differentiated cellular behavior remains poorly understood, mostly due to our inability to directly and precisely measure cell cycle state. In order to characterize cell cycle state live, we have adapted a cyclin dependent kinase (CDK) biosensor for in vivo use. We demonstrate here using a combination of spinning disk confocal and adaptive optics lattice light sheet microscopy the use of this biosensor to quantify lineage-specific differences between cycling cells, and to examine the link between quiescence and differentiation in the roundworm nematode, Caenorhabditis elegans and zebrafish. Unlike other live cell imaging tools (e.g., FUCCI), we show that DHB can be used to distinguish between actively cycling cells in the G1 phase of the cell cycle and quiescent or terminally differentiated cells exited in G0. Additionally, we have used this tool to investigate cell cycle regulation during morphogenesis and regeneration in zebrafish and through a multi-lab collaboration are exploring the use of this biosensor across the animal kingdom. Thus, we provide here a new resource to study the control and timing of the cell cycle during cell migration, fate specification and differentiation.

Rebecca Adikes1, Nicholas Palmisano1, Abraham Khorman1, Michael Martinez1, Jayson Smith1, Taylor Medwig-Kinney1, Tian-Ming Fu3, Maria Jose Gacha Garay1, Ononnah Ahmed1, Nicholas Weeks1, Nuri Kim1, Simeiyun Liu1, Rob Morabito1, Wan Zhang1, Eric Betzig3, Ariel Pani2, Benjamin Martin1, David Matus1
1) Stony Brook University, Stony Brook, NY 2) University of Virginia, Charlottesville, VA 3) Janelia Research Campus, Ashburn, VA