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      Seasonal Dynamics in the Chemistry and Structure of the Fat Bodies of Bumblebee Queens

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          Abstract

          Insects’ fat bodies are responsible for nutrient storage and for a significant part of intermediary metabolism. Thus, it can be expected that the structure and content of the fat body will adaptively change, if an insect is going through different life stages. Bumblebee queens belong to such insects as they dramatically change their physiology several times over their lives in relation to their solitary overwintering, independent colony foundation stage, and during the colony life-cycle ending in the senescent stage. Here, we report on changes in the ultrastructure and lipid composition of the peripheral fat body of Bombus terrestris queens in relation to seasonal changes in the queens’ activity. Six life stages are defined and evaluated in particular: pharate, callow, before and after hibernation, egg-laying, and senescence. Transmission electron microscopy revealed that the fat body contained two main cell types–adipocytes and oenocytes. Only adipocytes reveal important changes related to the life phase, and mostly the ration between inclusion and cytoplasm volume varies among particular stages. Both electron microscopy and chemical analyses of lipids highlighted seasonal variability in the quantity of the stored lipids, which peaked prior to hibernation. Triacylglycerols appeared to be the main energy source during hibernation, while the amount of glycogen before and after hibernation remained unchanged. In addition, we observed that the representation of some fatty acids within the triacylglycerols change during the queen’s life. Last but not least, we show that fat body cell membranes do not undergo substantial changes concerning phospholipid composition in relation to overwintering. This finding supports the hypothesis that the cold-adaptation strategy of bumblebee queens is more likely to be based on polyol accumulation than on the restructuring of lipid membranes.

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          Fat metabolism in insects.

          The study of fat metabolism in insects has received considerable attention over the years. Although by no means complete, there is a growing body of information about dietary lipid requirements, and the absolute requirement for sterol is of particular note. In this review we (a) summarize the state of understanding of the dietary requirements for the major lipids and (b) describe in detail the insect lipid transport system. Insects digest and absorb lipids similarly to vertebrates, but with some important differences. The hallmark of fat metabolism in insects centers on the lipid transport system. The major lipid transported is diacylglycerol, and it is carried by a high-density lipoprotein called lipophorin. Lipophorin is a reusable shuttle that picks up lipid from the gut and delivers it to tissues for storage or utilization without using the endocytic processes common to vertebrate cells. The mechanisms by which this occurs are not completely understood and offer fruitful areas for future research.
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            Host specific social parasites (Psithyrus) indicate chemical recognition system in bumblebees.

            Semiochemicals influence many aspects of insect behavior, including interactions between parasites and their hosts. We studied the chemical recognition system of bumblebees (Bombus) by examining the cuticular hydrocarbon cues of 14 species, including five species of social parasites, known as cuckoo bees (subgenus Psithyrus). We found that bumblebees possess species-specific alkene positional isomer profiles that are stable over large geographical regions and are mimicked by three host-specific cuckoo parasites. In three host-cuckoo associations where mimicry is poor, possibly due to recent host shifts, these cuckoos produce dodecyl acetate a known chemical repellent that allows the cuckoos to invade their host colonies. Our findings indicate cuckoos use two chemical mechanisms, mimicry and repellents, to invade their hosts, and this may reflect different stages of an ongoing dynamic arms race.
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              Seasonal acquisition of chill tolerance and restructuring of membrane glycerophospholipids in an overwintering insect: triggering by low temperature, desiccation and diapause progression.

              Adults of the insect Pyrrhocoris apterus acquire chill tolerance through the process of autumnal acclimatization. Field and laboratory experiments were conducted to separate the triggering effects of low temperatures, desiccation and diapause progression on the physiological characteristics related to chill tolerance with emphasis on the restructuring of glycerophospholipid (GPL) composition. Changes in relative proportions of major molecular species of glycerophosphoethanolamines (GPEtns) and glycerophosphocholines (GPChols) in thoracic muscle and fat body tissues were followed using HPLC coupled to electrospray ionisation mass spectrometry. The increase in relative proportion of 1-palmitoyl-2-linoleyl-sn-GPEtn at the expense of 1,2-dilinoleyl-sn-GPChol was the most prominent feature of the complex change observed in both tissues during autumnal acclimatization in the field. The relative proportion of total GPEtns increased, while the proportion of total GPChols decreased. The relative proportion of unsaturated fatty acyls slightly decreased. A similar restructuring response was seen during acclimatization in the field and cold acclimation in the laboratory. By contrast, the GPL changes related to desiccation and diapause progression were relatively small, differed qualitatively from the cold-acclimation response, and were accompanied with no increase of chill tolerance. Other features of autumnal acclimatization, i.e. depression of supercooling capacity and accumulation of polyhydric alcohols, were also triggered solely by low temperatures.

                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                11 November 2015
                2015
                : 10
                : 11
                : e0142261
                Affiliations
                [1 ]Agricultural Research, Ltd., Troubsko, Czech Republic
                [2 ]Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic
                [3 ]Institute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budějovice, Czech Republic
                [4 ]Faculty of Forestry and Wood Sciences, Czech University of Life Sciences, Prague, Czech Republic
                University of Graz, AUSTRIA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist. A.V. and O.K. are employed by Agricultural Research, Ltd. There are no patents, products in development or marketed products to declare. This declaration does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.

                Conceived and designed the experiments: AV OK JS IV. Performed the experiments: AV EK MK JS PJ OK. Analyzed the data: AV AT JS MK EK IV. Contributed reagents/materials/analysis tools: AV EK MK JS PJ OK. Wrote the paper: AV AT EK JS IV.

                Article
                PONE-D-15-22265
                10.1371/journal.pone.0142261
                4641598
                26559946
                a765bd8c-7960-4daf-afb0-0c3d7e719b7f
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 22 May 2015
                : 20 October 2015
                Page count
                Figures: 4, Tables: 1, Pages: 14
                Funding
                This work was supported by the Czech Science Foundation (14-04291S); http://www.gacr.cz/; IV OK AV, Institute of Organic Chemistry and Biochemistry (61388963); http://www.uochb.cz/web/structure/31.html; IV AT EK MK PJ, Internal Grant Agency of Faculty of Forestry and Wood Sciences (20134359); http://www.fld.czu.cz/cs/; JS, and Agricultural Research, Ltd. The funder provided support in the form of salaries for authors [A.V., O.K.], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.
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