title: Production and characterization of astrocyte-derived human apolipoprotein E isoforms from immortalized astrocytes and their interactions with amyloid-beta.
abstract: The apolipoprotein E (apoE) genotype is an important genetic risk factor for Alzheimer's disease (AD). In the central nervous system (CNS), most apoE is produced by astrocytes and is present in unique high-density lipoprotein (HDL)-like particles that have distinct properties from apoE derived from other sources. To develop an efficient system to produce astrocyte-derived apoE in large quantities, we produced and characterized immortalized cell lines from primary astrocyte cultures derived from human APOE knock-in mice. APOE2, APOE3, and APOE4 expressing cell lines were established that secrete apoE in HDL-like particles at similar levels, cholesterol composition, and size as those produced by primary astrocytes. In physiological buffers, astrocyte-secreted apoE3 and E4 associated equally well with amyloid-beta. Under the same conditions, only a small fraction of A beta formed sodium dodecyl sulfate (SDS)-stable complexes with apoE (E3 > E4). These immortalized astrocytes will be useful for studying mechanisms underlying the isoform-specific effects of apoE in the CNS.
authors.affiliation: Department of Neurology, Washington University School of Medicine, St. Louis, MO 63130, USA.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Morikawa
authors.forename: John D
authors.initials: JD
authors.lastname: Fryer
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Sullivan
authors.forename: Erin A
authors.initials: EA
authors.lastname: Christopher
authors.forename: Suzanne E
authors.initials: SE
authors.lastname: Wahrle
authors.forename: Ronald B
authors.initials: RB
authors.lastname: DeMattos
authors.forename: Mark A
authors.initials: MA
authors.lastname: O'Dell
authors.forename: Anne M
authors.initials: AM
authors.lastname: Fagan
authors.forename: Hilal A
authors.initials: HA
authors.lastname: Lashuel
authors.forename: Thomas
authors.initials: T
authors.lastname: Walz
authors.forename: Kiyofumi
authors.initials: K
authors.lastname: Asai
authors.forename: David M
authors.initials: DM
authors.lastname: Holtzman
mesh_terms: D016229:Amyloid beta-Peptides / Q000378:metabolism*; D000818:Animals; D001057:Apolipoproteins E / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D001253:Astrocytes / Q000378:metabolism* / Q000648:ultrastructure; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D006624:Hippocampus / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D020033:Protein Isoforms / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure
chemical_list: D016229:Amyloid beta-Peptides; D001057:Apolipoproteins E; D020033:Protein Isoforms