title: Ubiquitin-like Atg8 protein is expressed during autophagy and the encystation process in Naegleria gruberi.
abstract: Members of the Naegleria genus are free-living amoebae, and the only pathogenic specie described to date for humans is N. fowleri. However, as the complete genome of this specie has not been reported, non-pathogenic N. gruberi is employed to describe molecular pathways in N. fowleri. Regardless, certain mechanisms, such as autophagy, have not yet been characterized in N. gruberi. Autophagy is involved in different cellular processes in some protozoa, including the recycling of unnecessary organelles, development, and cell differentiation. In this work, we characterized autophagy in N. gruberi using the specific inducer rapamycin. The formation of autophagy vacuoles in treated trophozoites was observed by ultrastructural analysis, and real time quantitative PCR demonstrated overexpression of the atg8 gene. In addition, we detected an increase in the vacuolar acidification of treated amoebae using the LysoTracker. Finally, confocal microscopy was utilized to identify Atg8 protein signal in the cytoplasm of N. gruberi trophozoites induced with rapamycin and even in trophozoites induced to encyst. In conclusion, N. gruberi possesses an Atg8 protein homolog that is overexpressed during the autophagic mechanism induced by rapamycin and also during encystation of this free-living amoeba.
authors: Department of Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies of the National Polytechnic Institute, Av. IPN 2508, Col. San Pedro Zacatenco, 07360, Mexico City, Mexico.
authors: Roberto
authors: R
authors: Cárdenas-Zúñiga
authors: Multidisciplinary Laboratory of Research, Military School of Graduate in Health, University of the Army and Air Force, Cerrada de Palomas S/N, Col. Lomas de San Isidro, 11620, Mexico City, Mexico.|Laboratory of Molecular Biomedicine, Institutional Program of Molecular Biomedicine, National School of Medicine and Homeopathy, St. Guillermo Massieu H. 239, Col. La escalera, 07320, Mexico City, Mexico.
authors: Virginia
authors: V
authors: Sánchez-Monroy
authors: Department of Genetics and Molecular Biology, Center for Research and Advanced Studies of the National Polytechnic Institute, Av. IPN 2508, Col. San Pedro Zacatenco, 07360, Mexico City, Mexico.
authors: Rosa María
authors: RM
authors: Bermúdez-Cruz
authors: Department of Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies of the National Polytechnic Institute, Av. IPN 2508, Col. San Pedro Zacatenco, 07360, Mexico City, Mexico.
authors: Mario Alberto
authors: MA
authors: Rodríguez
authors: Department of Cell Biology, Center for Research and Advanced Studies of the National Polytechnic Institute, Av. IPN 2508, Col. San Pedro Zacatenco, 07360, Mexico City, Mexico.
authors: Jesús
authors: J
authors: Serrano-Luna
authors: Department of Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies of the National Polytechnic Institute, Av. IPN 2508, Col. San Pedro Zacatenco, 07360, Mexico City, Mexico. mineko@cinvestav.mx.
authors: Mineko
authors: http://orcid.org/0000-0003-2248-0220
authors: M
authors: Shibayama
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001343:Autophagy / Q000502:physiology*; D000071190:Autophagy-Related Protein 8 Family / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology*; D009253:Naegleria / Q000648:ultrastructure; D020123:Sirolimus / Q000494:pharmacology; D053058:Trophozoites; D025801:Ubiquitin
chemical_list: D000900:Anti-Bacterial Agents; D000071190:Autophagy-Related Protein 8 Family; D025801:Ubiquitin; D020123:Sirolimus
keywords: Atg8 protein; Autophagy; Encystation; Naegleria gruberi; Rapamycin