title: The internal architecture of leukocyte lipid body organelles captured by three-dimensional electron microscopy tomography.
abstract: Lipid bodies (LBs), also known as lipid droplets, are complex organelles of all eukaryotic cells linked to a variety of biological functions as well as to the development of human diseases. In cells from the immune system, such as eosinophils, neutrophils and macrophages, LBs are rapidly formed in the cytoplasm in response to inflammatory and infectious diseases and are sites of synthesis of eicosanoid lipid mediators. However, little is known about the structural organization of these organelles. It is unclear whether leukocyte LBs contain a hydrophobic core of neutral lipids as found in lipid droplets from adipocytes and how diverse proteins, including enzymes involved in eicosanoid formation, incorporate into LBs. Here, leukocyte LB ultrastructure was studied in detail by conventional transmission electron microscopy (TEM), immunogold EM and electron tomography. By careful analysis of the two-dimensional ultrastructure of LBs from human blood eosinophils under different conditions, we identified membranous structures within LBs in both resting and activated cells. Cyclooxygenase, a membrane inserted protein that catalyzes the first step in prostaglandin synthesis, was localized throughout the internum of LBs. We used fully automated dual-axis electron tomography to study the three-dimensional architecture of LBs in high resolution. By tracking 4 nm-thick serial digital sections we found that leukocyte LBs enclose an intricate system of membranes within their "cores". After computational reconstruction, we showed that these membranes are organized as a network of tubules which resemble the endoplasmic reticulum (ER). Our findings explain how membrane-bound proteins interact and are spatially arranged within LB "cores" and support a model for LB formation by incorporating cytoplasmic membranes of the ER, instead of the conventional view that LBs emerge from the ER leaflets. This is important to understand the functional capabilities of leukocyte LBs in health and during diverse diseases in which these organelles are functionally involved.
authors.affiliation: Laboratory of Cellular Biology, Department of Biology, Federal University of Juiz de Fora, UFJF, Juiz de Fora, MG, Brazil.
authors.forename: Rossana C N
authors.initials: RC
authors.lastname: Melo
authors.forename: Guillherme F
authors.initials: GF
authors.lastname: Paganoti
authors.forename: Ann M
authors.initials: AM
authors.lastname: Dvorak
authors.forename: Peter F
authors.initials: PF
authors.lastname: Weller
mesh_terms: D017667:Adipocytes / Q000166:cytology; D055032:Electron Microscope Tomography; D004804:Eosinophils / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D021621:Imaging, Three-Dimensional; D007425:Intracellular Membranes / Q000000981:diagnostic imaging / Q000378:metabolism; D008055:Lipids; D008565:Membrane Proteins / Q000378:metabolism; D015388:Organelles / Q000378:metabolism / Q000648:ultrastructure*; D014463:Ultrasonography
chemical_list: D008055:Lipids; D008565:Membrane Proteins
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