Summary
The capacity for polyol synthesis by larvae of Eurosta solidaginis was evaluated under aerobic versus anoxic (N2 gas atmosphere) conditions. Glycerol production occurred readily in aerobic larvae at 13°C. Under anoxic conditions, however, net glycerol accumulation was only 57% of the aerobic value after 18 d, but the total hydroxyl equivalents available for cryoprotection were balanced by the additional synthesis of sorbitol. The efficiency of carbon conversion to polyols was much lower in anaerobic larvae. The ATP requirement of glycerol biosynthesis necessitated a 22% greater consumption of carbohydrate, when anaerobic and resulted in the accumulation of equimolar amounts of l-lactate and l-alanine as fermentative end products. The ratio of polyols produced to glycolytic end products formed was consistent with the use of the hexose monophosphate shunt to generate the reducing equivalents needed for cryoprotectant synthesis. A comparable experiment analyzed sorbitol synthesis at 3°C under aerobic versus anoxic conditions. Sorbitol synthesis was initiated more rapidly in anaerobic larvae, and the final sorbitol levels attained after 18 d were 60% higher than in aerobic larvae. The enhanced sorbitol output under anoxia may be due to an obligate channeling of a high percentage of total carbon flow through the hexose monophosphate shunt at 3°C. Carbon processed in this way generates NADPH which, along with the NADH output of glycolysis, must be reoxidized if anaerobic ATP synthesis is to continue. Redox balance within the hexose monophosphate shunt is maintained through NADPH consumption in the synthesis of sorbitol.
References
Baust JG (1983) Protective agents: regulation of synthesis. Cryobiology 20: 357–364
Baust JG, Morrissey RE (1977) Strategies of low temperature adaptation. Proc Int Cong Entomol XV: 173–184
Baust JG, Rojas RR, Hamilton MD (1985) Life at low temperatures: representative insect adaptations. Cryo Lett 6: 199–210
Bergmeyer HU, Gruber W, Gutman I (1974) d-Sorbitol. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Academic Press, New York, pp 1323–1326
Churchill TA, Storey KB (1989) Metabolic corrrelates to glycerol biosynthesis in a freze-avoiding insect, Epiblema scudderiana. J Comp Physiol B 159: 461–472
Conradi-Larsen EM, Sømme L (1973) The overwintering of Pelophila borealis Payk. II. Aerobic and anaerobic metabolism. Nor Entomol Tidsskr 20: 325–332
Duman JG (1980) Factors involved in the overwintering survival of the freeze-tolerant beetle, Dendroides canadensis. J Comp Physiol 136: 53–59
Eggstein M, Kuhlmann E (1974) Triglycerides and glycerol: determination after alkaline hydrolysis. In: Bergymeyer HU (ed) Methods of enzymatic analysis. Academic Press, New York, pp 1825–1831
Furusawa T, Shikata M, Yamashita O (1982) Temperature dependent sorbitol utilization in diapause eggs of the silkworm, Bombyx mori. J Comp Physiol 147: 21–26
Gäde G (1984) Anaerobic energy metabolism. In: Hofmann KH (ed) Environmental physiology and biochemistry of insects. Springer, Berlin Heidelberg New York, pp 119–136
Hochachka PW, Somero GN (1984) Biochemical adaptation. Princeton University Press, Princeton
Hue L, Rider MH (1987) Role of fructose-2,6-bisphosphate in the control of glycolysis in mammalian tissues. Biochem J 245: 313–324
Lowry OH, Passonneau JV (1972) A flexible system of enzymatic analysis. Academic Press, New York
Meyer SGE (1978) Effects of heat, cold, anaerobiosis and inhibitors on metabolite concentrations in larvae of Callitroga macellaria. Insect Biochem 6: 471–477
Miller LK, Smith JS (1975) Production of threitol and sorbitol by an adult insect: association with freezing tolerance. Nature 258: 519–520
Morrissey RE, Baust JG (1976) The ontogeny of cold tolerance in the gall fly, Eurosta solidaginis. J Insect Physiol 22: 431–437
Rickards J, Kelleher MJ, Storey KB (1987) Strategies of freeze avoidance in larvae of the goldenrod gall moth, Epiblema scudderiana: winter profiles of a natural population. J Insect Physiol 33:443–450
Ring RA, Tesar D (1981) Adaptations to cold in Canadian arctic insects. Cryobiology 18:199–211
Rojas RR, Lee RE, Luu TA, Baust JG (1983) Temperature dependence-independence of antifreeze turnover in Eurosta solidaginis (Fitch). J Insect Physiol 29:865–869
Rojas RR, Lee RE, Baust JG (1986) Relationship of environmental water content to glycerol accumulation in the freezing tolerant larvae of Eurosta solidaginis (Fitch). Cryo Lett 7:234–245
Schaftingen E van (1984) d-Fructose-2,6-bisphosphate. In: Bermeyer HU (ed) Methods of enzymatic analysis. Verlag Chemie, Weinheim, pp 335–341
Sømme L (1967) The effect of temperature and anoxia on hemolymph composition and supercooling in three overwintering insects. J Insect Physiol 13:805–814
Sømme L (1982) Supercooling and winter survival in terrestrial arthropods. Comp Biochem Physiol A 73:519–543
Storey KB (1982) Phosphofructokinase from the overwintering gall fly larva, Eurosta solidaginis control of cryoprotectant polyol synthesis. Insect Biochem 12:501–505
Storey KB (1982) Metabolism and bound water in overwintering insects. Cryobiology 20:365–379
Storey KB (1985) Metabolic biochemistry of insect flight. In: Gilles R (ed) Circulation, respiration and metabolism. Springer, Berlin Heidelberg New York, pp 193–207
Storey JM Storey KB (1983) Regulation of cryoprotectant metabolism in the overwintering gall fly larva, Eurosta solidaginis: temperature control of glycerol and sorbitol levels. J Comp Physiol 149:495–502
Storey JM, Storey KB (1985) Freezing and cellular metabolism in the gall fly larva, Eurosta solidaginis. J Comp Physiol B 155:333–337
Storey JM, Storey KB (1986) Winter survival of the gall fly larva, Eurosta solidaginis: profiles of fuel reserves and cryoprotectants in a natural population. J Insect Physiol 32:549–556
Storey KB, Storey JM (1988) Freeze-tolerance in animals. Physiol Rev 68:27–84
Storey KB, Storey JM (1988) Biochemistry of cryoprotectants. In: Denlinger D, Lee RE (eds) Insects at low temperatures. John Wiley & Sons, New York, in press
Tsumuki H, Rojas RR, Storey KB, Baust JG (1987) The fate of [14C]glucose during cold-hardening in Eurosta solidaginis (Fitch). Insect Biochem 17:347–352
Wasylyk JM, Tice AR, Baust JG (1988) Partial glass formation: a novel mechanism of insect cryoprotection. Cryobiology 25:451–458
Wood FE, Nordin JH (1980) Activation of the hexose monophosphate shunt during cold-induced glycerol accumulation by Protophormia terranovae. Insect Biochem 10:87–93
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Storey, J.M., Storey, K.B. Carbon balance and energetics of cyooprotectant synthesis in a freeze-tolerant insect: responses to perturbation by anoxia. J Comp Physiol B 160, 77–84 (1990). https://doi.org/10.1007/BF00258765
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00258765