TY - JOUR
T1 - Ubiquitin-like Atg8 protein is expressed during autophagy and the encystation process in Naegleria gruberi
AU - Cárdenas-Zúñiga, Roberto
AU - Sánchez-Monroy, Virginia
AU - Bermúdez-Cruz, Rosa María
AU - Rodríguez, Mario Alberto
AU - Serrano-Luna, Jesús
AU - Shibayama, Mineko
N1 - Publisher Copyright:
© 2016, Springer-Verlag Berlin Heidelberg.
PY - 2017/1/1
Y1 - 2017/1/1
N2 - 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.
AB - 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.
KW - Atg8 protein
KW - Autophagy
KW - Encystation
KW - Naegleria gruberi
KW - Rapamycin
UR - http://www.scopus.com/inward/record.url?scp=84992702766&partnerID=8YFLogxK
U2 - 10.1007/s00436-016-5293-x
DO - 10.1007/s00436-016-5293-x
M3 - Artículo
C2 - 27796560
SN - 0932-0113
VL - 116
SP - 303
EP - 312
JO - Parasitology Research
JF - Parasitology Research
IS - 1
ER -