Quantitative assessment of the nanoanatomy of the contractile vacuole complex inTrypanosoma cruzi

Author:

Augusto Ingrid1ORCID,Girard-Dias Wendell12,Schoijet Alejandra34,Alonso Guillermo Daniel34,Portugal Rodrigo V56,de Souza Wanderley178,Jimenez Veronica9ORCID,Miranda Kildare178ORCID

Affiliation:

1. Laboratório de Ultraestrutura Celular Hertha Meyer, Centro de Pesquisa em Medicina de Precisão, Instituto de Biofísica Carlos Chagas Filho and Centro Nacional de Biologia Estrutural e Bioimagem, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil

2. Plataforma de Microscopia Eletrônica Rudolf Barth, Instituto Oswaldo Cruz – Fiocruz, Rio de Janeiro, Brazil

3. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular “Dr. Héctor N. Torres”, Buenos Aires, Argentina

4. Departamento de Fisiología, Biología Molecular y Celular, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina

5. Laboratório Nacional de Nanotecnologia, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, Brazil

6. Programa de Biotecnologia, Universidade Federal do ABC, Santo André, Brazil

7. Instituto Nacional de Ciência e Tecnologia em Biologia Estrutural e Bioimagem – Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil

8. Centro Multiusuário para Análise de Fenômenos Biomédicos, Universidade do Estado do Amazonas, Manaus, Brazil

9. Department of Biological Sciences, College of Natural Sciences and Mathematics, California State University Fullerton, Fullerton, CA, USA

Abstract

Trypanosoma cruziuses various mechanisms to cope with osmotic fluctuations during infection, including the remodeling of organelles such as the contractile vacuole complex (CVC). Little is known about the morphological changes of the CVC during pulsation cycles occurring upon osmotic stress. Here, we investigated the structure–function relationship between the CVC and the flagellar pocket domain where fluid discharge takes place—the adhesion plaque—during the CVC pulsation cycle. Using TcrPDEC2 and TcVps34 overexpressing mutants, known to have low and high efficiency for osmotic responses, we described a structural phenotype for the CVC that matches their corresponding physiological responses. Quantitative tomography provided data on the volume of the CVC and spongiome connections. Changes in the adhesion plaque during the pulsation cycle were also quantified and a dense filamentous network was observed. Together, the results suggest that the adhesion plaque mediates fluid discharge from the central vacuole, revealing new aspects of the osmoregulatory system inT. cruzi.

Funder

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro

Ministerio da Saude

Publisher

Life Science Alliance, LLC

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