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Death of PC12 cells and hippocampal neurons induced by adenoviral‐mediated FAD human amyloid precursor protein gene expression
Author(s) -
Luo JinJun,
Wallace William,
Riccioni Teresa,
Ingram Donald K.,
Roth George S.,
Kusiak John W.
Publication year - 1999
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/(sici)1097-4547(19990301)55:5<629::aid-jnr10>3.0.co;2-y
Subject(s) - hippocampal formation , amyloid precursor protein , microbiology and biotechnology , amyloid β , programmed cell death , gene , neuroscience , gene expression , biology , cancer research , apoptosis , medicine , alzheimer's disease , pathology , disease , biochemistry
We used adenoviral‐mediated gene transfer of human amyloid precursor proteins (h‐APPs) to evaluate the role of various h‐APPs in causing neuronal cell death. We were able to infect PC12 cells with very high efficiency because ≈90% of the cells were cytochemically positive for β‐galactosidase activity when an adenoviral vector containing LacZ cDNA was used to infect cells. Cells infected with adenovirus containing h‐APP cDNA showed high‐level transcription and expression of h‐APP as measured by reverse transcriptase–polymerase chain reaction and Western immunoblot analyses, respectively. Intracellular and extracellular levels of h‐APP were elevated approximately 17‐ and 24‐fold in cultures infected with recombinant adenovirus containing wild‐type mutant and 13‐ and 17‐fold with V642F mutant. No elevation in h‐APP was seen in cultures infected with antisense h‐APP or null adenovirus. H‐APP levels were maximal 3 days after infection. Overexpression of V642F mutant h‐APP in PC12 cells and hippocampal neurons resulted in about a twofold increase in death compared with overexpression of wild‐type h‐APP. These results demonstrate the usefulness of recombinant adenoviral mediated gene transfer in cell culture studies and suggest that overexpression of a familial Alzheimer's disease mutant APP may be toxic to neuronal cells. J. Neurosci. Res. 55:629–642, 1999 © 1999 Wiley‐Liss, Inc.