title: Phospholipase D2: a pivotal player modulating RBL-2H3 mast cell structure.
abstract: The current study examined the role of PLD2 in the maintenance of mast cell structure. Phospholipase D (PLD) catalyzes hydrolysis of phosphatidylcholine to produce choline and phosphatidic acid (PA). PLD has two isoforms, PLD1 and PLD2, which vary in expression and localization depending on the cell type. The mast cell line RBL-2H3 was transfected to overexpress catalytically active (PLD2CA) and inactive (PLD2CI) forms of PLD2. The results of this study show that PLD2CI cells have a distinct star-shaped morphology, whereas PLD2CA and RBL-2H3 cells are spindle shaped. In PLD2CI cells, the Golgi complex was also disorganized with dilated cisternae, and more Golgi-associated vesicles were present as compared with the PLD2CA and RBL-2H3 cells. Treatment with exogenous PA led to the restoration of the wild-type Golgi complex phenotype in PLD2CI cells. Conversely, treatment of RBL-2H3 and PLD2CA cells with 1% 1-Butanol led to a disruption of the Golgi complex. The distribution of acidic compartments, including secretory granules and lysosomes, was also modified in PLD2CI cells, where they concentrated in the perinuclear region. These results suggest that the PA produced by PLD2 plays an important role in regulating cell morphology in mast cells.
authors.affiliation: Department of Cell and Molecular Biology and Bioagents Pathogenic, Faculdade de Medicina de Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil.
authors.forename: Claudia Maria Meirelles
authors.initials: CM
authors.lastname: Marchini-Alves
authors.forename: Liliana Martos
authors.initials: LM
authors.lastname: Nicoletti
authors.forename: Vivian Marino
authors.initials: VM
authors.lastname: Mazucato
authors.forename: Lorena Brito
authors.initials: LB
authors.lastname: de Souza
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Hitomi
authors.forename: Cleidson de Pádua
authors.initials: Cde P
authors.lastname: Alves
authors.forename: Maria Celia
authors.initials: MC
authors.lastname: Jamur
authors.forename: Constance
authors.initials: C
authors.lastname: Oliver
mesh_terms: D000818:Animals; D002460:Cell Line; D006056:Golgi Apparatus / Q000201:enzymology / Q000648:ultrastructure; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008247:Lysosomes / Q000201:enzymology / Q000648:ultrastructure; D008407:Mast Cells / Q000166:cytology* / Q000201:enzymology / Q000648:ultrastructure; D010712:Phosphatidic Acids / Q000494:pharmacology; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D022142:Secretory Vesicles / Q000201:enzymology / Q000648:ultrastructure
chemical_list: D007527:Isoenzymes; D010712:Phosphatidic Acids; D012333:RNA, Messenger; C105606:phospholipase D2; D010739:Phospholipase D