Acidocalcisomes as Calcium- and Polyphosphate-Storage Compartments during Embryogenesis of the Insect Rhodnius prolixus Stahl 英文参考文献.docVIP

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Acidocalcisomes as Calcium- and Polyphosphate-Storage Compartments during Embryogenesis of the Insect Rhodnius prolixus Stahl 英文参考文献.doc

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Acidocalcisomes as Calcium- and Polyphosphate-Storage Compartments during Embryogenesis of the Insect Rhodnius prolixus Stahl 英文参考文献

AcidocalcisomesasCalcium-andPolyphosphate-Storage CompartmentsduringEmbryogenesisoftheInsect RhodniusprolixusStahl IsabelaRamos1,2,FabioGomes1,CarolinaM.Koeller1,KatsuharuSaito4,NortonHeise1 ,Hatisaburo Masuda2,RobertoDocampo5,WanderleydeSouza1,3,EdnildoA.Machado1,3*.,KildareMiranda 1,3 . * 1IntitutodeBiof?′sicaCarlosChagasFilho,UniversidadeFederaldoRiodeJaneiro,RiodeJaneiro,Brazil,2InstitutodeBioqu?′micaMe′dica,UniversidadeFederaldoRiode Janeiro, Rio de Janeiro, Brazil, 3Diretoria de Programas, Instituto Nacional de Metrologia Normalizac?a?o e Qualidade Industrial, Xere′m, Brazil, 4Faculty of Agriculture, ShinshuUniversity,Minamiminowa,Nagano,Japan,5DepartmentofCellularBiology,CenterforTropicalandEmergingGlobalDiseases,UniversityofGeorgia,Athens, Georgia,UnitedStatesofAmerica Abstract Background: The yolk of insect eggs is a cellular domain specialized in the storage of reserve components for embryo development.Thereservemacromoleculesarestoredindifferentorganellesandtheirinteractionswiththeembryocellsare mostly unknown. Acidocalcisomes are lysosome-related organelles characterized by their acidic nature, high electron density and large content of polyphosphate bound to several cations. In this work, we report the presence of acidocalcisome-likeorganellesineggsoftheinsectvectorRhodniusprolixus. Methodology/Principalfindings:Characterizationoftheelementalcompositionofelectron-densevesiclesbyelectronprobe X-raymicroanalysisrevealeda compositionsimilartothatpreviously describedforacidocalcisomes.Following subcellular fractionation experiments, fractions enriched in acidocalcisomes were obtained and characterized. Immunofluorescence + showedthatpolyphosphatepolymersandthevacuolarprotontranslocatingpyrophosphatase(V-H -PPase,consideredasa markerforacidocalcisomes)arefoundinthesamevesiclesandthattheseorganellesaremainlylocalizedintheeggcortex. Polyphosphatequantificationshowedthatacidocalcisomescontainasignificantamountofpolyphosphatedetectedatday-0 eggs.Elem