Electrically stimulated induction of hsp70 gene expression in mouse astroglia and fibroblast cells.
Mouse astroglial cells, which were cultured on an electrode, were found responsive to an electric stimulation of sine wave potential in enhancing hsp70 mRNA resulting from an activation of hsp70 gene expression. On the basis of this finding, electrically responsive cells were established by transfecting mouse 3T3-L1 cells with a constructed plasmid encoding hsp70 promoter and the firefly luciferase gene. A stable cell line has been established through selection of heat-stimulated luciferase expression. A 1-h electric stimulation of the cells resulted in activation of luciferase expression, which was confirmed to produce an increase in light emission. The sequential pattern of the electrically stimulated expression of luciferase was found different from that of the heat stimulation. Furthermore, the promoter was activated depending on the potential and duration of the stimulation applied. Consequently, the electric stimulation has proven effective on activating hspP70 promoter. This cell line is feasible in expressing the gene of interest by electrical stimulation, which lead us to construct environment responsive cells in general.
Yanagida
Y
Y
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Yokohama, Japan.
Mizuno
A
A
Motegi
T
T
Kobatake
E
E
Aizawa
M
M
D016475:3T3 Cells; D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D002478:Cells, Cultured; D004558:Electric Stimulation; D004566:Electrodes; D005347:Fibroblasts / Q000378:metabolism*; D015870:Gene Expression; D016762:Genes, fos / Q000235:genetics; D016758:Genes, jun / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D051379:Mice; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis
D018840:HSP70 Heat-Shock Proteins; D012333:RNA, Messenger