02229nas a2200265   4500008004600000245013400046210006900180300001600249490000700265520128500272653002201557653003201579653002501611653001201636653003001648653003101678653003501709653004101744653002801785653002701813653003101840100002001871700002201891856005001913       Submitted                   eng d     00aOleic acid is elevated in cell membranes during rapid cold-hardening and pupal diapause in the flesh fly, Sarcophaga crassipalpis0 aOleic acid is elevated in cell membranes during rapid coldharden  a1073 - 10820 v523 aThe integrity of cellular membranes is critical to the survival of insects at low temperatures, thus an advantage is conferred to insects that can adjust their composition of membrane fatty acids (FAs). Such changes contribute to homeoviscous adaption, a process that allows cellular membranes to maintain a liquid-crystalline state at temperatures that are potentially low enough to cause the membrane to enter the gel state and thereby lose its ability to maintain homeostasis. Flesh flies (Sarcophaga crassipalpis) were subjected to two experimental conditions that elicit low temperature tolerance: rapid cold-hardening and diapause. FAs were isolated and analyzed using gas chromatography-mass spectrometry. FAs changed in response to both rapid cold-hardening and diapause. In response to rapid cold-hardening (8 h at 4 degrees C), the proportion of oleic acid (18:1n-9) in pharate adults increased from 30% to 47% of the total FA pool. The proportion of almost every other FA was reduced. By entering diapause, pupae experienced an even greater increase in oleic acid proportion, to 58% of the total FA pool. Oleic acid not only promotes membrane fluidity at low temperature but also allows the cell membrane to maintain a liquid crystalline state if temperatures increase.10a*Cold Temperature10aAcclimatization/*physiology10aAnalysis of Variance10aAnimals10aCell Membrane/*metabolism10aChromatography, Thin Layer10aDiptera/metabolism/*physiology10aGas Chromatography-Mass Spectrometry10aHibernation/*physiology10aOleic Acid/*metabolism10aPupa/metabolism/physiology1 aMichaud, M., R.1 aDenlinger, D., L.  uhttps://sarcophagidae.myspecies.info/node/54502479nas a2200253   4500008004600000245019500046210006900241300001400310490000800324520153600332653002901868653002201897653002801919653001201947653003501959653004101994653002302035653002502058653003302083653001702116100002002133700002202153856005002175       Submitted                   eng d     00aShifts in the carbohydrate, polyol, and amino acid pools during rapid cold-hardening and diapause-associated cold-hardening in flesh flies (Sarcophaga crassipalpis): a metabolomic comparison0 aShifts in the carbohydrate polyol and amino acid pools during ra  a753 - 7630 v1773 aFlesh 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 beta-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.10a*Carbohydrate Metabolism10a*Cold Temperature10aAmino Acids/*metabolism10aAnimals10aDiptera/metabolism/*physiology10aGas Chromatography-Mass Spectrometry10aModels, Biological10aPolymers/*metabolism10aPrincipal Component Analysis10aTime Factors1 aMichaud, M., R.1 aDenlinger, D., L.  uhttps://sarcophagidae.myspecies.info/node/46702350nas a2200313   4500008004600000245014400046210006900190300001400259490000900273520122600282653003001508653001201538653002101550653002301571653003901594653003501633653004201668653003001710653003101740653003701771653003201808653002201840653005001862653002101912100001101933700002001944700002201964856005001986       Submitted                   eng d     00aRapid elevation of Inos and decreases in abundance of other proteins at pupal diapause termination in the flesh fly Sarcophaga crassipalpis0 aRapid elevation of Inos and decreases in abundance of other prot  a663 - 6680 v17943 aWe analyzed changes in brain proteins 24 h after pupal diapause termination in Sarcophaga crassipalpis by a combination of 2-dimensional gel electrophoresis and mass spectrometry. The proteome analysis revealed significant changes in 20 proteins, 11 of which represented &gt;or=2.5-fold changes. Three proteins were present only in the brains of diapausing pupae. Among the most abundant proteins that showed a change, 1 was more abundant, 7 were less abundant, and 2 were absent following diapause termination. The protein that increased in abundance following diapause termination showed highest identity to myo-inositol-1-phosphate synthase (Inos). Proteins that decreased at diapause termination included those showing highest identities to fatty acid binding protein, CG2331-PA, twinstar, catalase, and a histone. Proteins absent at diapause termination included ribosomal protein L17A and one unnamed protein. An increase of Inos protein level was confirmed using Western blot analysis. Attempts to terminate diapause by injection of several Inos-related metabolites failed, thus suggesting that the elevation of Inos at diapause termination is downstream of the physiological regulation that initiates development.10aAdaptation, Physiological10aAnimals10aBrain/metabolism10aDatabases, Protein10adevelopment/*metabolism/physiology10aDiptera/*metabolism/physiology10aElectrophoresis, Gel, Two-Dimensional10aGene Expression Profiling10aGene Expression Regulation10aInositol/metabolism/pharmacology10aInsect Proteins/*metabolism10aMass Spectrometry10aMyo-Inositol-1-Phosphate Synthase/*metabolism10aPupa/growth &amp1 aLi, A.1 aMichaud, M., R.1 aDenlinger, D., L.  uhttps://sarcophagidae.myspecies.info/node/284