ApoE synthesis in human neuroblastoma cells.
Apolipoprotein E (apoE) is associated with the two hallmarks of Alzheimer's disease: A beta deposits and neurofibrillary tangles. ApoE synthesis was detected in astrocytes by in situ hybridization but was not detected in neurons. Nevertheless, different studies on apoE immunoreactivity reported the presence of apoE in neurons of Alzheimer, control, and necrosis pontisubicular brains. In this study, we addressed the question of potential synthesis of apoE in neurons and its possible involvement in or in response to pathological conditions. To this purpose, we have studied human neuronal cell lines (SY 5Y and Kelly cells) originating from neuroblastoma. Using monoclonal and polyclonal antibodies, a 32-kDa band was detected in SY 5Y and Kelly cells, before and after NGF differentiation. Two-dimensional gel electrophoresis analysis showed a typical profile of apoE spots resolved to the exact isoelectric points. By reverse transcription-polymerase chain reaction experiments, we demonstrated the presence of apoE mRNA in these cell lines. SY 5Y cells synthesized the apoE3 variant, whereas Kelly cells expressed both apoE3 and apoE4 isoforms, corroborating the two-dimensional gel results. These results suggested that apoE synthesis could occur in human neuronal cell lines under certain conditions.
Dupont-Wallois
L
L
INSERM U422, Lille, France.
Soulié
C
C
Sergeant
N
N
Wavrant-de Wrieze
N
N
Chartier-Harlin
M C
MC
Delacourte
A
A
Caillet-Boudin
M L
ML
D000911:Antibodies, Monoclonal; D001057:Apolipoproteins E / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015415:Biomarkers; D015153:Blotting, Western; D003599:Cytoskeleton / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression; D006801:Humans; D009447:Neuroblastoma; D009457:Neuroglia / Q000737:chemistry / Q000378:metabolism; D009474:Neurons / Q000737:chemistry / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D014407:Tumor Cells, Cultured / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001057:Apolipoproteins E; D015415:Biomarkers; D012333:RNA, Messenger; D016875:tau Proteins