Shifts in the carbohydrate, polyol, and amino acid pools during rapid cold-hardening and diapause-associated cold-hardening in flesh flies (Sarcophaga crassipalpis): a metabolomic comparison

Abstract

Flesh flies can enhance their cold hardiness by entering a photoperiod-induced pupal diapause or by a temperature-induced rapid cold-hardening process. To determine whether the same or different metabolites are involved in these two responses, derivatized polar extracts from flesh flies subjected to these treatments were examined using gas chromatography–mass spectrophotometry (GC–MS). This metabolomic approach demonstrated that levels of metabolites involved in glycolysis (glycerol, glucose, alanine, pyruvate) were elevated by both treatments. Metabolites elevated uniquely in response to rapid cold-hardening include glutamine, cystathionine, sorbitol, and urea while levels of β-alanine, ornithine, trehalose, and mannose levels were reduced. Rapid cold-hardening also uniquely perturbed the urea cycle. In addition to the elevated metabolites shared with rapid cold-hardening, leucine concentrations were uniquely elevated during diapause while levels of a number of other amino acids were reduced. Pools of two aerobic metabolic intermediates, fumarate and citrate, were reduced during diapause, indicating a reduction of Krebs cycle activity. Principal component analysis demonstrated that rapid cold-hardening and diapause are metabolically distinct from their untreated, non-diapausing counterparts. We discuss the possible contribution of each altered metabolite in enhancing the overall cold hardiness of the organism, as well as the efficacy of GC–MS metabolomics for investigating insect physiological systems.

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Abbreviations

ANOVA:

Analysis of variance

GC–MS:

Gas chromatography–mass spectrometry

h:

hour

NMR:

Nuclear magnetic resonance

PCA:

Principal component analysis

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Acknowledgments

The authors of this manuscript thank Richard Sessler of the Ohio State Campus Chemical Instruments Center and Amelia Brown for their technical contributions to this manuscript. The authors also appreciate the reviewers of this article for their useful suggestions and comments. This study was funded by the National Science Foundation (#IOB-0416720) and complies with all federal, state, and local laws.

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Correspondence to M. Robert Michaud.

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Communicated by H.V. Carey.

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Michaud, M.R., Denlinger, D.L. Shifts in the carbohydrate, polyol, and amino acid pools during rapid cold-hardening and diapause-associated cold-hardening in flesh flies (Sarcophaga crassipalpis): a metabolomic comparison. J Comp Physiol B 177, 753–763 (2007). https://doi.org/10.1007/s00360-007-0172-5

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Keywords

  • Insect
  • Diapause
  • Amino acid
  • Polyol
  • Metabolism