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

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.

Automated summary

Trichomonas vaginalis is a parasite causing trichomoniasis in the human urogenital tract. When subjected to stressful conditions, it can differentiate into cyst-like stages showing resistance to unfavorable conditions. Research indicates that during this differentiation process, the mRNA of the meiotic recombinase Dmc1 is upregulated, suggesting potential cryptic sexual events in the cyst-like stages of T. vaginalis.