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DEVELOPMENTAL PHYSIOLOGY AND PREGNANCY

Hypoxia induces major effects on cell cycle kinetics and protein expression in Drosophila melanogaster embryos

Departments of ¹Pediatrics (Division of Respiratory Medicine) and ²Neuroscience, Albert Einstein College of Medicine, New York, New York; and ³Department of Genetics, Boyer Center for Molecular Medicine and ⁴The Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, Connecticut

Submitted 3 August 2004 ; accepted in final form 13 October 2004

Hypoxia induces a stereotypic response in Drosophila melanogaster embryos: depending on the time of hypoxia, embryos arrest cell cycle activity either at metaphase or just before S phase. To understand the mechanisms underlying hypoxia-induced arrest, two kinds of experiments were conducted. First, embryos carrying a kinesin-green fluorescent protein construct, which permits in vivo confocal microscopic visualization of the cell cycle, showed a dose-response relation between O₂ level and cell cycle length. For example, mild hypoxia (PO₂ 55 Torr) had no apparent effect on cell cycle length, whereas severe hypoxia (PO₂ 25–35 Torr) or anoxia (PO₂ = 0 Torr) arrested the cell cycle. Second, we utilized Drosophila embryos carrying a heat shock promoter driving the string (cdc25) gene (HS-STG3), which permits synchronization of embryos before the start of mitosis. Under conditions of anoxia, we induced a stabilization or an increase in the expression of several G1/S (e.g., dE2F1, RBF2) and G2/M (e.g., cyclin A, cyclin B, dWee1) proteins. This study suggests that, in fruit fly embryos, 1) there is a dose-dependent relationship between cell cycle length and O₂ levels in fruit fly embryos, and 2) stabilized cyclin A and E2F1 are likely to be the mediators of hypoxia-induced arrest at metaphase and pre-S phase.

cyclin A; E2F1; oxygen deprivation; stabilization of expression

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