Co-expression of beta-amyloid precursor protein (betaAPP) and apolipoprotein E in cell culture: analysis of betaAPP processing.
Apolipoprotein E (ApoE) is the major genetic risk factor for Alzheimer's disease (AD). The ApoE4 allele is associated with earlier disease onset and greater cerebral deposition of the amyloid beta peptide (Abeta), the major constituent of senile (amyloid) plaques. The molecular mechanism underlying these effects of ApoE4 remains unclear; ApoE alleles could have different influences on Abeta production, extracellular aggregation, or clearance. Because the missense mutations on chromosomes 14 and 21 that cause familial forms of AD appear to lead to increased secretion of Abeta, it is important to determine whether ApoE4 has a similar effect. Here, we have examined the effects of all three ApoE alleles on the processing of betaAPP and the secretion of Abeta in intact cells. We established neural (HS683 human glioma) and non-neural (Chinese hamster ovary) cell culture systems that constitutively secrete both ApoE and Abeta at concentrations like those in human cerebrospinal fluid. betaAPP metabolites, generated in the presence of each ApoE allele, were analysed and quantified by two methods: immunoprecipitation and phosphorimaging, and ELISA. We detected no consistent allele-specific effects of ApoE on betaAPP processing in either cell type. Our data suggest that the higher amyloid burden found in AD subjects expressing ApoE4 is not due to increased amyloidogenic processing of betaAPP, in contrast to findings in AD linked to chromosome 14 or 21. These co-expressing cell lines will be useful in the further search for the effects of ApoE on Abeta aggregation or clearance under physiologically relevant conditions.
Biere
A L
AL
Center for Neurologic Diseases, Harvard Medical School, Brigham and Women's Hospital, Boston, MA 02115, USA.
Ostaszewski
B
B
Zhao
H
H
Gillespie
S
S
Younkin
S G
SG
Selkoe
D J
DJ
D000544:Alzheimer Disease / Q000469:parasitology; D016564:Amyloid beta-Protein Precursor / Q000032:analysis*; D000818:Animals; D001057:Apolipoproteins E / Q000032:analysis*; D001483:Base Sequence; D015153:Blotting, Western; D001921:Brain / Q000473:pathology; D001923:Brain Chemistry; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D002883:Chromosomes, Human, Pair 14; D003001:Cloning, Molecular; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D011233:Precipitin Tests
D016564:Amyloid beta-Protein Precursor; D001057:Apolipoproteins E