Effects of temperature on juvenile Dungeness crab,<i>Metacarcinus magister</i>(Dana): survival, moulting, and mTOR signalling and neuropeptide gene expression in eyestalk ganglia, moulting gland (Y-organ), and heart

Abstract

Mechanistic target of rapamymcin (mTOR) is a highly conserved protein kinase that controls cellular protein synthesis and energy homeostasis. We hypothesize that mTOR integrates intrinsic signals (moulting hormones) and extrinsic signals (thermal stress) to regulate moulting and growth in decapod crustaceans. The effects of temperature on survival, moulting and mRNA levels of mTOR signalling genes (<i>Mm-Rheb</i>, <i>Mm-mTOR</i>, <i>Mm-AMPKα</i>, <i>Mm-S6K</i> and <i>Mm-AKT</i>) and neuropeptides (<i>Mm-CHH</i> and <i>Mm-MIH</i>) were quantified in juvenile <i>Metacarcinus magister</i> Crabs at different moult stages (12, 19 or 26 days postmoult) were transferred from ambient temperature (∼15°C) to temperatures between 5 and 30°C for up to 14 days. Survival was 97-100% from 5 to 20°C, but none survived at 25 or 30°C. Moult stage progression accelerated from 5 to 15°C, but did not accelerate further at 20°C. In eyestalk ganglia, <i>Mm-Rheb</i>, <i>Mm-AMPKα</i> and <i>Mm-AKT</i> mRNA levels decreased with increasing temperatures. <i>Mm-MIH</i> and <i>Mm-CHH</i> mRNA levels were lowest in the eyestalk ganglia of mid-premoult animals at 20°C. In the Y-organ, <i>Mm-Rheb</i> mRNA levels decreased with increasing temperature and increased during premoult, and were positively correlated with haemolymph ecdysteroid titre. In the heart, moult stage had no effect on mTOR signalling gene mRNA levels; only <i>Mm-Rheb</i>, <i>Mm-S6K</i> and <i>Mm-mTOR</i> mRNA levels were higher in intermoult animals at 10°C. These data suggest that temperature compensation of neuropeptide and mTOR signalling gene expression in the eyestalk ganglia and Y-organ contributes to regulate moulting in the 10 to 20°C range. The limited warm compensation in the heart may contribute to mortality at temperatures above 20°C.