Nondiapausing larvae of the flesh fly, Sarcophaga bullata, responded to several forms of short-term environmental stress (low temperature,
anoxia and desiccation) by accumulating
glycerol. Elevation of this
polyol, regardless of the type of stress that induced accumulation, conferred cold resistance: larvae with high
glycerol levels were 3-4 times more tolerant of a 2h exposure to -10 degrees C than unstressed larvae. Protection against low temperature injury, as well as
dehydration, was also attained by injection of exogenous
glycerol into third instar larvae. This artificially induced cold hardiness was only temporary: when
glycerol-injected larvae were exposed to -10 degrees C immediately after injection, survival was high, but none survived if they were injected and then held at 25 degrees C for 2 days before the -10 degrees C exposure. Larvae ligated behind the brain immediately after low temperature exposure failed to accumulate
glycerol, but
glycerol did accumulate in larvae ligated 6-24h after cold treatment, thus implying a critical role for the brain in initiating
glycerol production. Interestingly, a much shorter exposure (2h) to low temperature was sufficient to reduce the maximum rate of water loss. Collectively, these observations suggest that multiple pathways may be exploited in response to stress: one pathway is most likely associated with rapid cold hardening (RCH) which generates immediate protection, and a second pathway remains activated for a longer period to enhance the initial protection afforded by
glycerol.