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Biogenesis of Autophagosome in Trichomonas vaginalis during Macroautophagy Induced by Rapamycin‐treatment and Iron or Glucose Starvation Conditions
Author(s) -
HernándezGarcía Mar S.,
MirandaOzuna Jesús F. T.,
SalazarVillatoro Lizbeth,
VázquezCalzada Carlos,
ÁvilaGonzález Leticia,
GonzálezRobles Arturo,
OrtegaLópez Jaime,
Arroyo Rossana
Publication year - 2019
Publication title -
journal of eukaryotic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 1066-5234
DOI - 10.1111/jeu.12712
Subject(s) - autophagy , biology , autophagosome , atg8 , microbiology and biotechnology , cytoplasm , organelle , biogenesis , western blot , phagosome , vacuole , biochemistry , phagocytosis , apoptosis , gene
Autophagy is an adaptive response for cell survival in which cytoplasmic components and organelles are degraded in bulk under normal and stress conditions. Trichomonas vaginalis is a parasite highly adaptable to stress conditions such as iron ( IR ) and glucose restriction ( GR ). Autophagy can be traced by detecting a key autophagy protein (Atg8) anchored to the autophagosome membrane by a lipid moiety. Our goal was to perform a morphological and cellular study of autophagy in T. vaginalis under GR , IR , and Rapamycin (Rapa) treatment using TvAtg8 as a putative autophagy marker. We cloned tvatg8a and tvatg8b and expressed and purified rTvA tg8a and rTvA tg8b to produce specific polyclonal antibodies. Autophagy vesicles were detected by indirect immunofluorescence assays and confirmed by ultrastructural analysis. The biogenesis of autophagosomes was detected, showing intact cytosolic cargo. TvAtg8 was detected as puncta signal with the anti‐ rTvA tg8b antibody that recognized soluble and lipid‐associated TvAtg8b by Western blot assays in lysates from stress‐inducing conditions. The TvAtg8b signal co‐localized with the Cyto ID and lysotracker labeling (autolysosomes) that accumulated after E‐64d treatment in GR parasites. Our data suggest that autophagy induced by starvation in T. vaginalis results in the formation of autophagosomes for which TvAtg8b could be a putative autophagy marker.