Non-autonomous insulin signaling delays mitotic progression inC. elegansgermline stem and progenitor cells

Stem and progenitor cell mitosis is essential for tissue development and homeostasis. How these cells ensure proper chromosome segregation, and thereby maintain mitotic fidelity, in the complex physiological environment of a living animal is poorly understood. Here we use in situ live-cell imaging of C. elegans germline stem and progenitor cells (GSPCs) to ask how the signaling environment influences stem and progenitor cell mitosis in vivo . Through a candidate screen we identify a new role for the insulin/IGF receptor (IGFR), daf-2, during GSPC mitosis. Mitosis is delayed in daf-2/ IGFR mutants, and these delays require canonical, DAF-2/IGFR to DAF-16/FoxO insulin signaling, here acting cell non-autonomously from the soma. Interestingly, mitotic delays in daf-2 /IGFR mutants depend on the spindle assembly checkpoint but are not accompanied by a loss of mitotic fidelity. Correspondingly, we show that caloric restriction, which delays GSPC mitosis and compromises mitotic fidelity, does not act via the canonical insulin signaling pathway, and instead requires AMP-activated kinase (AMPK). Together this work demonstrates that GSPC mitosis is influenced by at least two genetically separable signaling pathways and highlights the importance of signaling networks for proper stem and progenitor cell mitosis in vivo . Author Summary Stem and progenitor cells drive tissue development and sustain adult tissue turnover by producing new daughter cells via cell division, the success of which depends on proper chromosome segregation during mitosis. Stem and progenitor cells perform mitosis in the complex environment of a living animal, yet relatively little is known about how events during mitosis are influenced by this in vivo context. In particular, whether signaling pathways that coordinate other aspects of stem and progenitor cell behavior with animal physiology also play a role during mitosis is poorly understood. Here we took advantage of the germline stem and progenitor cells of the model nematode C. elegans to address this question. Through live-cell imaging of germline stem and progenitor cell mitosis, we uncover a new role for the insulin signaling pathway. We find that reducing insulin signaling delays germline stem and progenitor cell mitosis, but, surprisingly, these delays are not accompanied by a loss of mitotic fidelity. In addition, we find that reducing insulin signaling in somatic tissues is sufficient to delay germline stem and progenitor mitosis, indicating that the pathway acts non-autonomously. Finally, while insulin signaling is known to link cell division with nutritional status in many species, we found that it did not mediate the effects of caloric restriction on germline stem and progenitor cell mitosis. Instead, caloric restriction acts via the conserved energy-sensing regulator AMPK. These results uncover new regulators of germline stem and progenitor cell mitosis and emphasize the importance of signaling pathways for proper stem and progenitor cell mitosis in vivo .