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. 2014 Feb 5;5:19. doi: 10.3389/fphys.2014.00019

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Copyright © 2014 Smith, Lamattina and Collins.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Effects of PI3K inhibitors (LY294002 and wortmannin), and MEK inhibitor (U0126) on ecdysone secretion. Prothoracic glands from day 3 fifth instar larvae were incubated for 20 min in medium with or without the designated inhibitor, followed by 60 min in one of the following treatments: No Inhibitor (Grace's medium); LY294002 (60 uM); Wortmannin (10 uM); U0126 (10 uM); Brain Extract (0.5 brain equivalents/30 ul); Brain Extract + LY294002; Brain Extract + Wortmannin; Brain Extract + U0126; PTTH (7 nM); or PTTH + LY294002. At the end of the 60 min incubation period, culture medium was assayed for ecdysone by radioimmunoassay. Ecdysone secretion by groups treated with U0126 was significantly lower than by groups in the respective no-inhibitor controls (asterisks) (ANOVA, Tukey–Kramer post-hoc test, significance p < 0.05). Sample sizes were as follows (glands per group): Control = 51; BE = 38; LY294002 = 36; BE + LY294002 = 27; U0126 = 4; BE + U0126 = 5; Wortmannin = 10; BE + Wortmannin = 12; PTTH = 19; PTTH + LY = 19.