Practicable recovery of plasmids carrying cDNAs encoding anti-apoptotic factors that had been introduced by particle bombardment into 5-week-old rat hippocampal slices

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. Copyright (C) 1999 Elsevier Science Ireland Ltd.