Death of PC12 cells and hippocampal neurons induced by adenoviral-mediated FAD human amyloid precursor protein gene expression
✍ Scribed by Jin-Jun Luo; William Wallace; Teresa Riccioni; Donald K. Ingram; George S. Roth; John W. Kusiak
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 746 KB
- Volume
- 55
- Category
- Article
- ISSN
- 0360-4012
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✦ Synopsis
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 D90% of the cells were cytochemically positive for -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 17and 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.