Eiichi Nagashima

Cell cycles in embryos of the silkworm, Bombyx mori: G2-arrest at diapause stage

SummaryIn the silkworm, Bombyx mori, diapause occurs at a specific embryonic stage, i.e. after formation of the germ band with cephalic lobes and telson and sequential mesoderm segmentation. As long as the eggs are incubated at 25° C, cell divisions and morphological development of the embryos cease. To examine changes in percentage of embryonic cells in the G1, S and G2 phases during embryogenesis, nuclear fractions were isolated from embryos, stained with propidium iodide and then subjected to flow cytometric analysis. The percentages of embryonic cells in G1, S and G2 were 10, 35 and 55%, respectively, at the stage of formation of cephalic lobes, whilst 98% of cells were in G2 at diapause stage. After termination of diapause by acclimation at 5° C or by a combination of chilling and HCl, cell division resumed in the embryos. During this period, the cells rapidly entered S phase through G1 from G2, suggesting that their G1 phase was short. In eggs in which diapause was averted by HCl-treatment after incubation at 25° C for 20 h after oviposition, embryonic development proceeded continuously for 9.5 days at 25° C until hatching. Along with this development, the G1 fraction increased to levels of about 90%. These results indicate that embryonic cells are arrested in G2 at diapause and suggest that, concomitant with further embryonic development, cell cycles become slower in proportion to an increasing length of G1. Finally, most of the cells may be arrested in G1, while there is only a small fraction of cells continuously cycling.