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Dynamic In vivo Imaging of NADPH Oxidase Gene Expression to Monitor its Involvement in Morphine's Actions
Author(s) -
XIA Min,
Hull Lynn C,
Thomas Cindy,
Gabra Bichoy H,
Li Ningjun,
Li PinLan,
Dewey William L
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1125.13
Subject(s) - in vivo , luciferase , transfection , gene expression , microbiology and biotechnology , ex vivo , chemistry , biology , gene , biochemistry
The present study examined the applicability of using in vivo molecular imaging to monitor the daily expression of locally transfected NADPH oxidase 4 (Nox4) gene in periaqueductal gray (PAG) of mice. A mixture of luciferase expression plasmids and transfection reagent with vectors containing Nox4 was injected into the lateral ventricle at 2 mm rostral and 2 mm lateral with at a 45° angle from the bregma. Luciferase expression was monitored daily using an IVIS 200 in vivo imaging system. The in vivo bioluminescent signals were detectable as early as 24 hours after gene transfection into mouse brain and were maintained for 7‐days. On days 2 to 7, the antinociceptive efficacy of morphine was significantly increased in Nox4 vector‐injected compared to empty vector‐injected and naïve mice. After in vivo monitoring, the mice were euthanized and brain slices were obtained for ex vivo imaging. Strong signals of transfection of luciferase genes were seen in the PAG area. By real‐time RT‐PCR, Nox4 mRNA abundance was also found doubled in the dissected PAG, but not in the brain cortex. These results show that the in vivo imaging system used in these experiments provides a powerful tool for daily non‐invasive dynamic visualization of gene expression and will be useful for future studies of the role of other genes in drug effects in the brain. (Supported by DK54927, DA01647, DA020836 and HL89563)