Insulin-regulated growth of the prothoracic glands appears to play a critical role in timing the last larval molt, and hence metamorphosis. The present study examined insulin signaling in relation to the growth and secretory activity of prothoracic glands in the tobacco hornworm, Manduca sexta. As larvae feed during the first half of the final larval stage, the prothoracic glands grow and ecdysone secretory capacity increases. During this period of growth, we verified the presence of insulin receptor transcript in the prothoracic glands and demonstrated that the glands were responsive to insulin, as evidenced by the in vitro phosphorylation of signaling proteins in the insulin pathway such as Akt/protein kinase B and FOXO. It was predicted that starvation would reduce ecdysone secretion with concomitant changes in insulin signaling. To test this prediction, larvae were starved and changes were quantified in two nutritionally sensitive transcripts, insulin receptor and the translation inhibitor 4EBP. In glands from starved larvae, growth and ecdysone secretory capacity were reduced, and insulin receptor and 4EBP transcripts were increased. The latter changes would be expected to accompany starvation in conjunction with enhanced insulin sensitivity and reduced protein synthesis. Increased transcription of insulin receptor and 4EBP strongly suggest that nutritional deprivation reduces the secretion of endogenous insulin-like hormones. When injected with insulin, 4EBP levels in the prothoracic glands of starved larvae decreased. Thus, insulin appeared to correct starvation-induced deficits in glandular protein synthesis. However, insulin injection did not enhance ecdysone secretion. Thus, although the prothoracic glands are insulin-responsive and insulin-like hormones may promote glandular growth as larvae feed, the effects of nutritional depletion on steroidogenesis in Manduca cannot be explained solely by reduced insulin.
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