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Glucose uptake in the mammalian stages of Trypanosoma cruzi
Authors:Ariel M. Silber, Renata R. Tonelli, Camila G. Lopes, Narcisa Cunha-e-Silva, Ana Cl  udia T. Torrecilhas, Robert I. Schumacher, Walter Colli,Maria Jú  lia M. Alves
Affiliation:aDepartamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Av. Prof Lineu Prestes 748, CEP 05508-900, São Paulo, SP, Brazil;bInstituto de Biofisica Carlos Chagas Filho, CCS, Bloco G, subsolo, Universidade Federal do Rio de Janeiro, Cidade Universitária, 21949-900, Rio de Janeiro, RJ, Brazil
Abstract:Trypanosoma cruzi, the agent of Chagas’ disease, alternates between different morphogenetic stages that face distinct physiological conditions in their invertebrate and vertebrate hosts, likely in the availability of glucose. While the glucose transport is well characterized in epimastigotes of T. cruzi, nothing is known about how the mammalian stages acquire this molecule. Herein glucose transport activity and expression were analyzed in the three developmental stages present in the vertebrate cycle of T. cruzi. The infective trypomastigotes showed the highest transport activity (Vmax = 5.34 ± 0.54 nmol/min per mg of protein; Km = 0.38 ± 0.01 mM) when compared to intracellular epimastigotes (Vmax = 2.18 ± 0.20 nmol/min per mg of protein; Km = 0.39 ± 0.01 mM). Under the conditions employed no transport activity could be detected in amastigotes. The gene of the glucose transporter is expressed at the mRNA level in trypomastigotes and in intracellular epimastigotes but not in amastigotes, as revealed by real-time PCR. In both trypomastigotes and intracellular epimastigotes protein expression could be detected by Western blot with an antibody raised against the glucose transporter correlating well with the transport activity measured experimentally. Interestingly, anti-glucose transporter antibodies showed a strong reactivity with glycosome and reservosome organelles. A comparison between proline and glucose transport among the intracellular differentiation forms is presented. The data suggest that the regulation of glucose transporter reflects different energy and carbon requirements along the intracellular life cycle of T. cruzi.
Keywords:Trypanosoma cruzi   Hexose transporter   Glucose uptake   Intracellular stages
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