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Establishment of suitable reference genes for studying relative gene expression during the transition from trophozoites to cyst-like stages and first evidences of stress-induced expression of meiotic genes in Trichomonas vaginalis

Published online by Cambridge University Press:  08 April 2021

Juliana Figueiredo Peixoto ,
Daniele Graças dos Santos ,
Lupis Ribeiro ,
Vitor Silva Cândido de Oliveira ,
Rodrigo Nunes-da-Fonseca  and
José Luciano Nepomuceno-Silva Open the ORCID record for José Luciano Nepomuceno-Silva [Opens in a new window]
Juliana Figueiredo Peixoto
Affiliation:
Laboratório Integrado de Bioquímica Hatisaburo Masuda (LIBHM), Instituto de Biodiversidade e Sustentabilidade (NUPEM/UFRJ), Universidade Federal do Rio de Janeiro, Macaé/RJ, Brazil
Daniele Graças dos Santos
Affiliation:
Laboratório Integrado de Bioquímica Hatisaburo Masuda (LIBHM), Instituto de Biodiversidade e Sustentabilidade (NUPEM/UFRJ), Universidade Federal do Rio de Janeiro, Macaé/RJ, Brazil
Lupis Ribeiro
Affiliation:
Laboratório Integrado de Ciências Morfofuncionais (LICM), Instituto de Biodiversidade e Sustentabilidade (NUPEM/UFRJ), Universidade Federal do Rio de Janeiro, Macaé/RJ, Brazil
Vitor Silva Cândido de Oliveira
Affiliation:
Laboratório Integrado de Bioquímica Hatisaburo Masuda (LIBHM), Instituto de Biodiversidade e Sustentabilidade (NUPEM/UFRJ), Universidade Federal do Rio de Janeiro, Macaé/RJ, Brazil
Rodrigo Nunes-da-Fonseca
Affiliation:
Laboratório Integrado de Ciências Morfofuncionais (LICM), Instituto de Biodiversidade e Sustentabilidade (NUPEM/UFRJ), Universidade Federal do Rio de Janeiro, Macaé/RJ, Brazil
José Luciano Nepomuceno-Silva*
Affiliation:
Laboratório Integrado de Bioquímica Hatisaburo Masuda (LIBHM), Instituto de Biodiversidade e Sustentabilidade (NUPEM/UFRJ), Universidade Federal do Rio de Janeiro, Macaé/RJ, Brazil
*
Author for correspondence: José Luciano Nepomuceno-Silva, E-mail: nepomuc@yahoo.com
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Abstract

Trichomonas vaginalis is a parasite of the human urogenital tract and the causative agent of trichomoniasis, a sexually transmitted disease of worldwide importance. This parasite is usually found as a motile flagellated trophozoite. However, when subjected to stressful microenvironmental conditions, T. vaginalis trophozoites can differentiate into peculiar cyst-like stages, which exhibit notable physiological resistance to unfavourable conditions. Although well documented in morphological and proteomic terms, patterns of gene expression changes involved in the cellular differentiation into cyst-like stages are mostly unknown. The real-time reverse transcription polymerase chain reaction (RT-qPCR) is recognized as a sensitive and accurate method for quantification of gene expression, providing fluorescence-based data that are proportional to the amount of a target RNA. However, the reliability of relative expression studies depends on the validation of suitable reference genes, which RNAs exhibit a minimum of variation between tested conditions. Here, we attempt to determine suitable reference genes to be used as controls of invariant expression during cold-induced in vitro differentiation of T. vaginalis trophozoites into cyst-like forms. Furthermore, we reveal that the mRNA from the meiotic recombinase Dmc1 is upregulated during this process, indicating that cryptic sexual events may take place in cyst-like stages of T. vaginalis.

Keywords

Cyst-like formsDmc1quantitative PCRreference geneTrichomonas vaginalis
Type
Research Article
Information
Parasitology , Volume 148 , Issue 8 , July 2021 , pp. 934 - 946
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Copyright
Copyright © The Author(s), 2021. Published by Cambridge University Press

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Footnotes

*

Present address: Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro/RJ, Brazil.

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