Low GSK3β activity is required for insect diapause through responding to ROS/AKT signaling and down-regulation of Smad1/EcR/HR3 cascade.

Glycogen synthase kinase 3β (GSK3β) plays important roles in gene transcription, metabolism, apoptosis, development, and signal transduction. However, its role in the regulation of pupal diapause remains unclear. In this paper, we find that low GSK3β activity in brains of diapause-destined pupae of Helicoverpa armigera is caused by elevated AKT activity. In response to ROS, AKT phosphorylates GSK3β to decrease its activity. In developing pupal brains, GSK3β can activate the transcription factor Smad1, which binds to the promoter region of the ecdysone receptor (EcR) gene and increases its expression. In the presence of 20-hydroxyecdysone (20E), EcR can bind to USP and increase the expression of 20E-response genes, including HR3, for pupal-adult development. In contrast, high levels of ROS in brains of diapause-destined pupae up-regulate p-AKT, which in turn decreases GSK3β activity. Low GSK3β activity causes low expression of EcR/HR3 via down-regulation of Smad1 activity, leading to diapause initiation. These results suggest that low GSK3β activity plays a key role in pupal diapause via ROS/AKT/GSK3β/Smad/EcR/HR3 signaling.