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Chip-calorimetric assessment of heat generation during Ca2+ uptake by digitonin-permeabilized Trypanosoma cruzi

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Abstract

Ca2+ signaling in trypanosomatids is an important component of energy metabolism regulation and therefore, cytosolic Ca2+ concentration is finely regulated by Ca2+ transport through the plasma membrane and Ca2+ uptake and release by intracellular organelles. To maintain intracellular Ca2+ homeostasis with different gradients of the ion within the cellular compartments, there is an energy cost and also energy dissipation in the form of heat. Using an innovative segmented fusion technique of a chip-calorimeter and CRISPR/Cas9 knockout (–KO) Trypanosoma cruzi cell lines, we evaluated the heat generation during Ca2+ uptake by digitonin-permeabilized T. cruzi epimastigotes, a system consisting of Ca2+ uptake predominantly by mitochondria and acidocalcisomes. We used three T. cruzi epimastigotes cell lines: control cells denominated scrambled, cells with the absence of the pyruvate dehydrogenase phosphatase (TcPDP-KO) and cells lacking mitochondrial Ca2+ uptake via the mitochondrial calcium uniporter (TcMCU-KO), that presented, in this respective order, decreasing rates and capacities of Ca2+ uptake. TcPDP-KO cells exhibited the lowest heat production following Ca2+ addition, which may be due to its lower mitochondrial oxidative phosphorylation capacity and lower ATP availability for acidocalcisomal Ca2+ uptake. Scrambled and TcMCU-KO cells exhibited similar Ca2+-induced heat effects, which correlates with a higher ATP-dependent acidocalcisomal Ca2+ uptake in these cells. Our results show evidences that mitochondrial Ca2+ transport via the uniporter is minimally heat dissipative, while ATPase pumps in acidocalcisomes possess a predominant contribution to the heat generated during Ca2+ uptake.

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Acknowledgements

This study was supported by the São Paulo Research Foundation (Fundação de Amparo à Pesquisa do Estado de São Paulo, FAPESP) visiting researcher Grant No. 2018/19976-1 to JL/AEV and FAEPEX Grant No. 519.292 to JL; FAPESP Grant No. 2017/17728-8 to AEV/RFC; and FAPESP postdoctoral fellowship to MRS/AEV (2017/05487-6).

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Authors and Affiliations

  1. Department of Pathology, Faculty of Medical Sciences, University of Campinas (UNICAMP), Campinas, SP, 13083-887, Brazil

    Marina Rincon Sartori, R. F. Castilho & A. E. Vercesi

  2. Institute of Physical Chemistry, TU Bergakademie Freiberg, Leipziger Str. 29, 09599, Freiberg, Germany

    J. Lerchner & F. Mertens

  3. Institute of Chemistry, University of Campinas (UNICAMP), Campinas, SP, 13083-861, Brazil

    P. O. Volpe

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  1. Marina Rincon Sartori
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by MRS and JL. The first draft of the manuscript was written by MRS and JL and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Marina Rincon Sartori.

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Sartori, M.R., Lerchner, J., Castilho, R.F. et al. Chip-calorimetric assessment of heat generation during Ca2+ uptake by digitonin-permeabilized Trypanosoma cruzi. J Therm Anal Calorim 147, 4611–4619 (2022). https://doi.org/10.1007/s10973-021-10862-8

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