title: Membrane cholesterol regulates lysosome-plasma membrane fusion events and modulates Trypanosoma cruzi invasion of host cells.
abstract: BACKGROUND
  Trypomastigotes of Trypanosoma cruzi are able to invade several types of non-phagocytic cells through a lysosomal dependent mechanism. It has been shown that, during invasion, parasites trigger host cell lysosome exocytosis, which initially occurs at the parasite-host contact site. Acid sphingomyelinase released from lysosomes then induces endocytosis and parasite internalization. Lysosomes continue to fuse with the newly formed parasitophorous vacuole until the parasite is completely enclosed by lysosomal membrane, a process indispensable for a stable infection. Previous work has shown that host membrane cholesterol is also important for the T. cruzi invasion process in both professional (macrophages) and non-professional (epithelial) phagocytic cells. However, the mechanism by which cholesterol-enriched microdomains participate in this process has remained unclear.
  
  
  RESULTS
  In the present work we show that cardiomyocytes treated with MβCD, a drug able to sequester cholesterol from cell membranes, leads to a 50% reduction in invasion by T. cruzi trypomastigotes, as well as a decrease in the number of recently internalized parasites co-localizing with lysosomal markers. Cholesterol depletion from host membranes was accompanied by a decrease in the labeling of host membrane lipid rafts, as well as excessive lysosome exocytic events during the earlier stages of treatment. Precocious lysosomal exocytosis in MβCD treated cells led to a change in lysosomal distribution, with a reduction in the number of these organelles at the cell periphery, and probably compromises the intracellular pool of lysosomes necessary for T. cruzi invasion.
  
  
  CONCLUSIONS
  Based on these results, we propose that cholesterol depletion leads to unregulated exocytic events, reducing lysosome availability at the cell cortex and consequently compromise T. cruzi entry into host cells. The results also suggest that two different pools of lysosomes are available in the cell and that cholesterol depletion may modulate the fusion of pre-docked lysosomes at the cell cortex.
authors.affiliation: Department of Morphology, Federal University of Minas Gerais, Belo Horizonte, Brazil.
authors.forename: Bárbara
authors.initials: B
authors.lastname: Hissa
authors.forename: Jacqueline G
authors.initials: JG
authors.lastname: Duarte
authors.forename: Ludmila F
authors.initials: LF
authors.lastname: Kelles
authors.forename: Fabio P
authors.initials: FP
authors.lastname: Santos
authors.forename: Helen L
authors.initials: HL
authors.lastname: del Puerto
authors.forename: Pedro H
authors.initials: PH
authors.lastname: Gazzinelli-Guimarães
authors.forename: Ana M
authors.initials: AM
authors.lastname: de Paula
authors.forename: Ubirajara
authors.initials: U
authors.lastname: Agero
authors.forename: Oscar N
authors.initials: ON
authors.lastname: Mesquita
authors.forename: Cristina
authors.initials: C
authors.lastname: Guatimosim
authors.forename: Egler
authors.initials: E
authors.lastname: Chiari
authors.forename: Luciana O
authors.initials: LO
authors.lastname: Andrade
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry*; D002478:Cells, Cultured; D002784:Cholesterol / Q000032:analysis*; D005089:Exocytosis; D008247:Lysosomes / Q000378:metabolism*; D008561:Membrane Fusion; D051379:Mice; D032383:Myocytes, Cardiac / Q000469:parasitology; D014349:Trypanosoma cruzi / Q000472:pathogenicity*
chemical_list: D002784:Cholesterol