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Interaction With the Extracellular Matrix Triggers Calcium Signaling in Trypanosoma cruzi Prior to Cell Invasion
Trypanosoma cruzi, the etiological agent of Chagas disease in humans, infects a wide variety of vertebrates. Trypomastigotes, the parasite infective forms, invade mammalian cells by a still poorly understood mechanism. Adhesion of tissue culture- derived trypomastigotes to the extracellular matrix (...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8521164/ https://www.ncbi.nlm.nih.gov/pubmed/34671568 http://dx.doi.org/10.3389/fcimb.2021.731372 |
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author | Manchola Varón, Nubia Carolina dos Santos, Guilherme Rodrigo R. M. Colli, Walter Alves, Maria Julia M. |
author_facet | Manchola Varón, Nubia Carolina dos Santos, Guilherme Rodrigo R. M. Colli, Walter Alves, Maria Julia M. |
author_sort | Manchola Varón, Nubia Carolina |
collection | PubMed |
description | Trypanosoma cruzi, the etiological agent of Chagas disease in humans, infects a wide variety of vertebrates. Trypomastigotes, the parasite infective forms, invade mammalian cells by a still poorly understood mechanism. Adhesion of tissue culture- derived trypomastigotes to the extracellular matrix (ECM) prior to cell invasion has been shown to be a relevant part of the process. Changes in phosphorylation, S-nitrosylation, and nitration levels of proteins, in the late phase of the interaction (2 h), leading to the reprogramming of both trypomastigotes metabolism and the DNA binding profile of modified histones, were described by our group. Here, the involvement of calcium signaling at a very early phase of parasite interaction with ECM is described. Increments in the intracellular calcium concentrations during trypomastigotes-ECM interaction depends on the Ca(2+) uptake from the extracellular medium, since it is inhibited by EGTA or Nifedipine, an inhibitor of the L-type voltage gated Ca(2+) channels and sphingosine-dependent plasma membrane Ca(2+) channel, but not by Vanadate, an inhibitor of the plasma membrane Ca(2+)-ATPase. Furthermore, Nifedipine inhibits the invasion of host cells by tissue culture- derived trypomastigotes in a dose-dependent manner, reaching 95% inhibition at 100 µM Nifedipine. These data indicate the importance of both Ca(2+) uptake from the medium and parasite-ECM interaction for host-cell invasion. Previous treatment of ECM with protease abolishes the Ca(2+) uptake, further reinforcing the possibility that these events may be connected. The mitochondrion plays a relevant role in Ca(2+) homeostasis in trypomastigotes during their interaction with ECM, as shown by the increment of the intracellular Ca(2+) concentration in the presence of Antimycin A, in contrast to other calcium homeostasis disruptors, such as Cyclopiazonic acid for endoplasmic reticulum and Bafilomycin A for acidocalcisome. Total phosphatase activity in the parasite decreases in the presence of Nifedipine, EGTA, and Okadaic acid, implying a role of calcium in the phosphorylation level of proteins that are interacting with the ECM in tissue culture- derived trypomastigotes. In summary, we describe here the increment of Ca(2+) at an early phase of the trypomastigotes interaction with ECM, implicating both nifedipine-sensitive Ca(2+) channels in the influx of Ca(2+) and the mitochondrion as the relevant organelle in Ca(2+) homeostasis. The data unravel a complex sequence of events prior to host cell invasion itself. |
format | Online Article Text |
id | pubmed-8521164 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-85211642021-10-19 Interaction With the Extracellular Matrix Triggers Calcium Signaling in Trypanosoma cruzi Prior to Cell Invasion Manchola Varón, Nubia Carolina dos Santos, Guilherme Rodrigo R. M. Colli, Walter Alves, Maria Julia M. Front Cell Infect Microbiol Cellular and Infection Microbiology Trypanosoma cruzi, the etiological agent of Chagas disease in humans, infects a wide variety of vertebrates. Trypomastigotes, the parasite infective forms, invade mammalian cells by a still poorly understood mechanism. Adhesion of tissue culture- derived trypomastigotes to the extracellular matrix (ECM) prior to cell invasion has been shown to be a relevant part of the process. Changes in phosphorylation, S-nitrosylation, and nitration levels of proteins, in the late phase of the interaction (2 h), leading to the reprogramming of both trypomastigotes metabolism and the DNA binding profile of modified histones, were described by our group. Here, the involvement of calcium signaling at a very early phase of parasite interaction with ECM is described. Increments in the intracellular calcium concentrations during trypomastigotes-ECM interaction depends on the Ca(2+) uptake from the extracellular medium, since it is inhibited by EGTA or Nifedipine, an inhibitor of the L-type voltage gated Ca(2+) channels and sphingosine-dependent plasma membrane Ca(2+) channel, but not by Vanadate, an inhibitor of the plasma membrane Ca(2+)-ATPase. Furthermore, Nifedipine inhibits the invasion of host cells by tissue culture- derived trypomastigotes in a dose-dependent manner, reaching 95% inhibition at 100 µM Nifedipine. These data indicate the importance of both Ca(2+) uptake from the medium and parasite-ECM interaction for host-cell invasion. Previous treatment of ECM with protease abolishes the Ca(2+) uptake, further reinforcing the possibility that these events may be connected. The mitochondrion plays a relevant role in Ca(2+) homeostasis in trypomastigotes during their interaction with ECM, as shown by the increment of the intracellular Ca(2+) concentration in the presence of Antimycin A, in contrast to other calcium homeostasis disruptors, such as Cyclopiazonic acid for endoplasmic reticulum and Bafilomycin A for acidocalcisome. Total phosphatase activity in the parasite decreases in the presence of Nifedipine, EGTA, and Okadaic acid, implying a role of calcium in the phosphorylation level of proteins that are interacting with the ECM in tissue culture- derived trypomastigotes. In summary, we describe here the increment of Ca(2+) at an early phase of the trypomastigotes interaction with ECM, implicating both nifedipine-sensitive Ca(2+) channels in the influx of Ca(2+) and the mitochondrion as the relevant organelle in Ca(2+) homeostasis. The data unravel a complex sequence of events prior to host cell invasion itself. Frontiers Media S.A. 2021-10-04 /pmc/articles/PMC8521164/ /pubmed/34671568 http://dx.doi.org/10.3389/fcimb.2021.731372 Text en Copyright © 2021 Manchola Varón, dos Santos, Colli and Alves https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cellular and Infection Microbiology Manchola Varón, Nubia Carolina dos Santos, Guilherme Rodrigo R. M. Colli, Walter Alves, Maria Julia M. Interaction With the Extracellular Matrix Triggers Calcium Signaling in Trypanosoma cruzi Prior to Cell Invasion |
title | Interaction With the Extracellular Matrix Triggers Calcium Signaling in Trypanosoma cruzi Prior to Cell Invasion |
title_full | Interaction With the Extracellular Matrix Triggers Calcium Signaling in Trypanosoma cruzi Prior to Cell Invasion |
title_fullStr | Interaction With the Extracellular Matrix Triggers Calcium Signaling in Trypanosoma cruzi Prior to Cell Invasion |
title_full_unstemmed | Interaction With the Extracellular Matrix Triggers Calcium Signaling in Trypanosoma cruzi Prior to Cell Invasion |
title_short | Interaction With the Extracellular Matrix Triggers Calcium Signaling in Trypanosoma cruzi Prior to Cell Invasion |
title_sort | interaction with the extracellular matrix triggers calcium signaling in trypanosoma cruzi prior to cell invasion |
topic | Cellular and Infection Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8521164/ https://www.ncbi.nlm.nih.gov/pubmed/34671568 http://dx.doi.org/10.3389/fcimb.2021.731372 |
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