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Dynamics of polymorphism of acidocalcisomes in<Emphasis Type="Italic"> Leishmania</Emphasis> parasites
Authors:Kildare?Miranda  Roberto?Docampo  Orlando?Grillo  Anderson?Franzen  Márcia?Attias  Anibal?Vercesi  Helmut?Plattner  Joachim?Hentschel  Email author" target="_blank">Wanderley?de?SouzaEmail author
Institution:(1) Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Av. Brigadeiro Trompovski, s/n., bloco G, Cidade Universitaria, CEP 21949-900 Rio de Janeiro, Brazil;(2) Laboratory of Molecular Parasitology and Center for Zoonoses Research, Department of Pathobiology, University of Illinois at Urbana, Champaign, IL, USA;(3) Lehrstuhl f. Zellbiologie/Ultrastrukturforschung, Fachbereich Biology, Universität Konstanz, Konstanz, Germany;(4) Laboratório de Bioenergética, Núcleo de Medicina e Cirugia Experimental, Universidade Estadual de Campinas, Campinas-SP, Brazil
Abstract:Growth of Leishmania mexicana amazonensis promastigotes in different culture media resulted in structurally and chemically different acidocalcisomes. When grown in SDM-79 medium, the promastigotes showed large spherical acidocalcisomes of up to 1.2 mgrm diameter distributed throughout the cell. X-ray microanalysis and elemental mapping of the organelles showed large amounts of oxygen, phosphorus, sodium, potassium, magnesium, calcium, and zinc. Immunofluorescence microscopy using antisera raised against a peptide sequence of the vacuolar-type proton pyrophosphatase of Arabidopsis thaliana that is conserved in the Leishmania enzyme, indicated localization in acidocalcisomes. When cells were transferred to Warrenrsquos medium, the acidocalcisomes transformed from spherical into branched tubular organelles. The labeling pattern of the vacuolar proton-pyrophosphatase, considered as a marker for the organelle, changed accompanying the structural changes of the acidocalcisomes, and the enzyme showed an apparently lower proton-transporting activity when measured in digitonin-permeabilized promastigotes. X-ray microanalysis and elemental mapping of these structures revealed the additional presence of iron. Together, the results reveal that the morphology and composition of acidocalcisomes are greatly influenced by the culture conditions.Electronic Supplementary Material Supplementary material is available in the online version of this article at
Keywords:Leishmania amazonensis  Acidocalcisomes  Elemental mapping  Vacuolar proton pyrophosphatase  Iron uptake
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