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.
Asoh
S
S
Department of Biochemistry and Cell Biology, Institute of Gerontology, Nippon Medical School, Kawasaki-city, Kanagawa-ken, Japan.
Mori
T
T
Ohta
S
S
D000367:Age Factors; D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics*; D019254:Genes, bcl-2 / Q000235:genetics*; D006624:Hippocampus / Q000166:cytology / Q000502:physiology; D010957:Plasmids / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014162:Transfection / Q000235:genetics / Q000379:methods*; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein
C494828:Bax protein, rat; C494813:Bcl2l1 protein, rat; D018076:DNA, Complementary; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein