Production and characterization of astrocyte-derived human apolipoprotein E isoforms from immortalized astrocytes and their interactions with amyloid-beta.
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.
Morikawa
Masayuki
M
Department of Neurology, Washington University School of Medicine, St. Louis, MO 63130, USA.
Fryer
John D
JD
Sullivan
Patrick M
PM
Christopher
Erin A
EA
Wahrle
Suzanne E
SE
DeMattos
Ronald B
RB
O'Dell
Mark A
MA
Fagan
Anne M
AM
Lashuel
Hilal A
HA
Walz
Thomas
T
Asai
Kiyofumi
K
Holtzman
David M
DM
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
D016229:Amyloid beta-Peptides; D001057:Apolipoproteins E; D020033:Protein Isoforms