Effects of temperature on survival, moulting, and expression of neuropeptide and mTOR signalling genes in juvenile Dungeness crab (Metacarcinus magister)

摘要

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 (Mm-Rheb, Mm-mTOR, Mm-AMPK alpha, Mm-S6K and Mm-AKT) and neuropeptides (Mm-CHH and Mm-MIH) were quantified in juvenile Metacarcinus magister. Crabs at different moult stages (12, 19 or 26 days postmoult) were transferred from ambient temperature (similar to 15 degrees C) to temperatures between 5 and 30 degrees C for up to 14 days. Survival was 97-100% from 5 to 20 degrees C, but none survived at 25 or 30 degrees C. Moult stage progression accelerated from 5 to 15 degrees C, but did not accelerate further at 20 degrees C. In eyestalk ganglia, Mm-Rheb, Mm-AMPKa and Mm-AKT mRNA levels decreased with increasing temperatures. Mm-M!H and Mm-CHH mRNA levels were lowest in the eyestalk ganglia of mid-premoult animals at 20 degrees C. In the Y-organ, Mm-Rheb 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 Mm-Rheb, Mm-S6K and Mm-mTOR mRNA levels were higher in intermoult animals at 10 degrees 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 degrees C range. The limited warm compensation in the heart may contribute to mortality at temperatures above 20 degrees C.