title: Identification of reggie-1 and reggie-2 as plasmamembrane-associated proteins which cocluster with activated GPI-anchored cell adhesion molecules in non-caveolar micropatches in neurons.
abstract: Neurons are believed to possess plasmalemmal microdomains and proteins analogous to the caveolae and caveolin of nonneuronal cells. Caveolae are plasmalemmal invaginations where activated glycosyl-phosphatidylinositol (GPI)-anchored proteins preferentially assemble and where transmembrane signaling may occur. Molecular cloning of rat reggie-1 and -2 (80% identical to goldfish reggie proteins) shows that reggie-2 is practically identical to mouse flotillin-1. Flotillin-1 and epidermal surface antigen (ESA) (flotillin-2) are suggested to represent possible membrane proteins in caveolae. Rat reggie-1 is 99% homologous to ESA in overlapping sequences but has a 49-amino-acid N-terminus not present in ESA. Antibodies (ABs) which recognize reggie-1 or -2 reveal that both proteins cluster at the plasmamembrane and occur in micropatches in neurons [dorsal root ganglia (DRGs), retinal ganglion, and PC-12 cells] and in nonneuronal cells. In neurons, reggie micropatches occur along the axon and in lamellipodia and filopodia of growth cones, but they do not occur in caveolae. By quantitative electronmicroscopic analysis we demonstrate the absence of caveolae in (anti-caveolin negative) neurons and show anti-reggie-1 immunogold-labeled clusters at the plasmamembrane of DRGs. When ABs against the GPI-anchored cell adhesion molecules (CAMs) F3 and Thy-1 are applied to live DRGs, the GPI-linked CAMs sequester into micropatches. Double immunofluorescence shows a colocalization of the CAMs with micropatches of anti-reggie antibodies. Thus, reggie-1 and reggie-2 identify sites where activated GPI-linked CAMs preferentially accumulate and which may represent noncaveolar micropatches (domains).
authors.affiliation: Department of Biology, University of Konstanz, Germany.
authors.forename: D M
authors.initials: DM
authors.lastname: Lang
authors.forename: S
authors.initials: S
authors.lastname: Lommel
authors.forename: M
authors.initials: M
authors.lastname: Jung
authors.forename: R
authors.initials: R
authors.lastname: Ankerhold
authors.forename: B
authors.initials: B
authors.lastname: Petrausch
authors.forename: U
authors.initials: U
authors.lastname: Laessing
authors.forename: M F
authors.initials: MF
authors.lastname: Wiechers
authors.forename: H
authors.initials: H
authors.lastname: Plattner
authors.forename: C A
authors.initials: CA
authors.lastname: Stuermer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000954:Antigens, Surface / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001253:Astrocytes / Q000378:metabolism / Q000648:ultrastructure; D051242:Caveolin 1; D022461:Caveolins; D015815:Cell Adhesion Molecules / Q000378:metabolism* / Q000648:ultrastructure; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003001:Cloning, Molecular; D029941:Fish Proteins; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006054:Goldfish; D020439:Growth Cones / Q000378:metabolism / Q000648:ultrastructure; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D016716:PC12 Cells; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012165:Retinal Ganglion Cells / Q000378:metabolism / Q000648:ultrastructure; D014617:Vacuoles / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D000954:Antigens, Surface; C495541:Cav1 protein, mouse; C495537:Cav1 protein, rat; D051242:Caveolin 1; D022461:Caveolins; D015815:Cell Adhesion Molecules; D029941:Fish Proteins; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C106354:flotillins; C104436:reggie-2 protein, Carassius auratus