Dynamic Control of Cell Cycle and Growth Coupling by Ecdysone, EGFR, and PI3K Signaling in Drosophila Histoblasts

12 pages, 7 figures.-- PMID: 19355788 [PubMed].-- Supporting information (Suppl. figures S1-S3, movies S1-S7) available at: http://biology.plosjournals.org/perlserv/?request=get-document&doi=10.1371/journal.pbio.1000079#toclink6Regulation of cell proliferation has been extensively studied in cultured cell systems that are characterized by coordinated growth and cell-cycle progression and relatively uniform cell size distribution. During the development of multicellular organisms, however, growth and division can be temporally uncoupled, and the signaling pathways that regulate these growth programs are poorly understood. A good model for analyzing proliferation control in such systems is the morphogenesis of the Drosophila adult abdominal epidermis by histoblasts. These cells undergo a series of temporally regulated transitions during which neither cell size nor division rate is constant. The proliferation of histoblasts during metamorphosis is uniquely amenable to clonal analysis in combination with live imaging. Thereby, we show that abdominal histoblasts, which grow while in G2 arrest during larval stages, enter a proliferative stage in the pupal period that is initiated by ecdysone-dependent string/Cdc25 phosphatase transcription. The proliferating histoblasts have preaccumulated stores of Cyclin E, which trigger an immediate S phase onset after mitosis. These rapid cell cycles lack a G1 phase and result in a progressive reduction of cell size. Eventually, the histoblasts proceed to a stage of slower proliferation that, in contrast to the preceding, depends on epidermal growth factor receptor (EGFR) signaling for progression through the G2/M transition and on insulin receptor/PI3K-mediated signaling for growth. These results uncover the developmentally programmed changes coupling the growth and proliferation of the histoblasts that form the abdominal epidermis of Drosophila. Histoblasts proceed through three distinct stages: growth without division, division without growth, and growth-coupled proliferation. Our identification of the signaling pathways and cell-cycle regulators that control these programs illustrates the power of in vivo time-lapse analyses after clone induction. It sets the stage for the comprehensive understanding of the coordination of cell growth and cell-cycle progression in complex multicellular eukaryotes.[Author summary] A fundamental issue in biology is the question of how the rate of cell division is coupled to cell growth. Coordination of these processes has been studied extensively in cultured cell systems but to a much lesser extent in intact organisms. To study this phenomenon in a physiological setting, we developed a methodology to observe and manipulate cell division and growth in a population of Drosophila abdominal cells called histoblasts. The various developmental stages of histoblast morphogenesis include exit from cell-cycle arrest, initially rapid growth in the absence of cell division, and subsequent coupling of proliferation and growth. We identified several critical developmental signaling pathways (including signaling via ecdysone, the EGF receptor, and PI 3-kinase) that regulate and coordinate cell growth and division cycles during these different types of cell-cycle phenomena. We propose that the internal logic of the Drosophila histoblast system may serve as a basic framework for understanding not only how coordinated cell growth and division operate in a number of other developmental contexts, but also how misregulation of cell growth and division occurs in contexts such as cancer cell populations.NN was supported by a Formación de Profesorado Universitario (FPU) fellowship from the Spanish Ministry of Education and Science (MEC) and CM by the "Juan de la Cierva" MEC program. Work in EM-B's lab is supported by grants of the DGICYT (MEC) and the European Union (EU) (WOUND-STREP Project).Peer reviewe