In vivo analysis of glucocorticoid-induced reporter gene expression using gene gun DNA delivery.
Glucocorticoid regulates various physiological processes via the activation and repression of gene expression. The anti-inflammatory effects and the adverse effects are believed to be dependent on the repression and the activation of genes, respectively. Reporter gene assay is a useful technique to separately evaluate these two functions and has been used for in vitro screening of novel ligands for the glucocorticoid receptor (GR). We report here the application of a reporter gene assay for the in vivo determination of the GR-mediated gene activation. A reporter plasmid containing glucocorticoid response elements was introduced to abdominal mouse skin using a gene gun. Administration of prednisolone induced the expression of the reporter gene, only when the GR expression plasmid was co-transfected with the reporter plasmid. Endogenous levels of corticosterone appeared to be negligible in this protocol. The dose response for this induction was comparable to those for the decreases in thymus weight and serum corticosterone. These results suggest that gene gun-mediated skin transfection enables the in vivo reporter gene assay and that this technique can be used to predict the potency of ligands for the GR-mediated gene activation.
Pharmacobioregulation Research Laboratory, Taiho Pharmaceutical Co, Ltd, Hanno, Saitama, Japan.
Kiyoshi
K
Tanigawa
Katsunao
K
Tanaka
Hidetaka
H
Nagase
Hidekazu
H
Miyake
Mamoru
M
Kjniwa
Koichi
K
Ikizawa
D000818:Animals; D019470:Biolistics / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000502:physiology; D017930:Genes, Reporter / Q000187:drug effects* / Q000502:physiology; D005938:Glucocorticoids / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D015533:Transcriptional Activation
D005938:Glucocorticoids