02334nas a2200277   4500008004600000245010100046210006900147300001400216490000800230520140100238653002201639653003201661653002501693653001201718653004401730653002901774653002401803653002401827653002101851653003801872653003401910653003101944100001501975700001601990856005002006       Submitted                   eng d     00aIn vivo and in vitro rapid cold-hardening protects cells from cold-shock injury in the flesh fly0 aIn vivo and in vitro rapid coldhardening protects cells from col  a611 - 6150 v1743 aThe capacity to undergo rapid the cold-hardening response (RCH) has been documented in diverse groups of insects and functions to protect against non-freezing cold injury and to preserve physiological performance in response to environmental cooling. The RCH response is remarkable for the rapidity of its induction; however the mechanism by which insects perceive cold and transduce this input at the cellular level has received little attention. To test the hypothesis that cells from isolated tissues can undergo RCH in response to cold, we assessed cell viability in four tissues that had undergone either RCH (0 degree C, 2 h followed by -8 degrees C, 2 h) or cold-shock (-8 degrees C, 2 h) both in vivo and in vitro from the adult flesh fly Sarcophaga crassipalpis (Diptera: Sarcophagidae) using fluorescent probes. Adult flies showed a significantly higher survival rate in the RCH group than in the cold-shocked group. Similarly, in all tissues tested, both in vivo and in vitro, RCH significantly improved cell survival compared with the respective cold-shocked groups. To our knowledge this is the first report to demonstrate that isolated cells and tissues from insects can undergo RCH. These results indicate that insect cells are capable of cold-sensing without neuroendocrine mediation; direct induction at the cellular level also helps to explain the swiftness of the RCH response.10a*Cold Temperature10aAcclimatization/*physiology10aAnalysis of Variance10aAnimals10aBody Temperature Regulation/*physiology10aCell Survival/physiology10aDiptera/*physiology10aFat Body/physiology10aFluorescent Dyes10aGastrointestinal Tract/physiology10aMalpighian Tubules/physiology10aSalivary Glands/physiology1 aYi, S., X.1 aLee, R., E.  uhttps://sarcophagidae.myspecies.info/node/77002229nas 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/54502476nas a2200277   4500008004600000245014300046210006900189300001600258490000800274520150900282653002201791653003201813653002401845653001201869653002301881653002801904653003601932653002201968653005401990653002002044653000902064653003602073100001702109700002202126856005002148       Submitted                   eng d     00ap38 MAPK is a likely component of the signal transduction pathway triggering rapid cold hardening in the flesh fly Sarcophaga crassipalpis0 ap38 MAPK is a likely component of the signal transduction pathwa  a3295 - 33000 v2103 aRapid cold hardening (RCH) is an adaptation enabling insects to quickly respond to low temperature, but little is known about the molecular events that trigger this response. In this study of the flesh fly Sarcophaga crassipalpis, we explore a possible role for mitogen-activated protein kinases (MAPKs) in the low temperature signaling that elicits RCH. We report that p38 MAPK from S. crassipalpis, which shows high cDNA sequence homology to p38 MAPKs from other insects and mammals, is rapidly activated at temperatures around 0 degrees C, temperatures that are most effective for inducing RCH. By contrast, low temperature does not activate either extracellular signal-regulated kinase (ERK) or Jun N-terminal kinase (JNK). An increase in phospho-p38 MAPK was observed within 10 min following exposure to 0 degrees C and reached its maximum level in 2 h. When flies were transferred from 0 to 25 degrees C, the level of phospho-p38 MAPK decreased immediately and reached trace levels by 3 h. Nondiapausing flies were much more responsive to p38 MAPK activation than cold-hardy diapausing pupae. Thus, p38 MAPK activation and RCH both show the same narrow ranges of temperature sensitivity, temporal profiles of activation and decay, and developmental specificity. These correlations suggest that p38 MAPK plays a potential role in regulating the induction of RCH. The p38 MAPK response was not dependent upon the brain, as evidenced by high activation in isolated abdomens exposed to low temperature.10a*Cold Temperature10aAcclimatization/*physiology10aAmino Acid Sequence10aAnimals10aCloning, Molecular10adevelopment/*physiology10aDiptera/*enzymology/growth &amp10aEnzyme Activation10ap38 Mitogen-Activated Protein Kinases/*metabolism10aPhosphorylation10aPupa10aSignal Transduction/*physiology1 aFujiwara, Y.1 aDenlinger, D., L.  uhttps://sarcophagidae.myspecies.info/node/470