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First published online March 2, 2006
Journal of Experimental Biology 209, 1016-1023 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02086
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Glycerol production in rainbow smelt (Osmerus mordax) may be triggered by low temperature alone and is associated with the activation of glycerol-3-phosphate dehydrogenase and glycerol-3-phosphatase

William R. Driedzic1,*, Kathy A. Clow1, Connie E. Short1 and K. Vanya Ewart2

1 Ocean Sciences Centre, Memorial University of Newfoundland, St John's, Newfoundland, Canada A1C 5S7
2 NRC institute for Marine Biosciences, 1411 Oxford Street, Halifax, Nova Scotia, Canada B3H 3Z1

* Author for correspondence (e-mail: wdriedzic{at}mun.ca)

Accepted 10 January 2006

Rainbow smelt (Osmerus mordax) accumulate high levels of glycerol in winter that serves as an antifreeze. Fish were subjected to controlled decreases in water temperature and levels of plasma glycerol, liver metabolites and liver enzymes were determined in order to identify control mechanisms for the initiation of glycerol synthesis. In two separate experiments, decreases in temperature from 8°C to 0°C over a period of 10–11 days resulted in increases in plasma glycerol from levels of less than 4 mmol l–1 to approximate mean levels of 40 (first experiment) and 150 mmol l–1 (second experiment). In a third experiment, decreases in temperature to –1°C resulted in plasma glycerol levels approaching 500 mmol l–1. The accumulation of glycerol could be driven in either December or March, thus eliminating decreasing photoperiod as a necessary cue for glycerol accumulation. Glycerol accumulation in plasma was associated with changes in metabolites in liver leading to increases in the mass action ratio across the reactions catalyzed by glycerol-3-phosphate dehydrogenase (GPDH) and glycerol-3-phosphatase (G3Pase). The maximal, in vitro activity of GPDH, increased twofold in association with a sharp increase in plasma glycerol level. The metabolite levels and enzyme activities provide complementary evidence that GPDH is a regulatory site in the low temperature triggered synthesis of glycerol. Indirect evidence, based on calculated rates of in vivo glycerol production by liver, suggests that G3Pase is a potential rate-limiting step. As well, transient increases in glyceraldehyde-3-phosphate dehydrogenase and alanine aminotransferase suggest that these sites are components of a suite of responses, in rainbow smelt liver, induced by low temperature.

Key words: rainbow smelt, osmerus mordax, glycerol, glycerol-3-phosphate dehydrogenase, glycerol-3-phosphatase, low temperature, freeze resistance


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