Practicable recovery of plasmids carrying cDNAs encoding anti-apoptotic factors that had been introduced by particle bombardment into 5-week old rat hippocampal slices. |
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Authors: | S Asoh T Mori S Ohta |
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Institution: | Department of Biochemistry and Cell Biology, Institute of Gerontology, Nippon Medical School, Kawasaki-city, Kanagawa-ken, Japan. |
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Abstract: | In the central nervous system, neuronal cells interact with glial cells and functionally differentiate, a process which can not be reproduced in cell culture. Identification of the novel factors involved in the growth and/or rescue of the differentiated neuronal cells has been impeded by a lack of methods for selecting the genes. In this study, hippocampal slices of a 5-week old rat were transiently introduced with plasmid DNA carrying anti-apoptotic rat bcl-2 or bcl-x cDNA by a particle-bombardment transfection procedure. The plasmid DNAs were expected not to be digested in living cells. Intact plasmid DNAs were recovered by PCR amplification from the slices with bcl-2 or bcl-x cDNAs but not from slices with empty vector or bax cDNA that promotes cell death. This study proposed that a technical combination of organotypic culture and particle-bombardment transfection is profitable for identifying novel genes that promote the survival of neuronal cells. |
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