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Temporal and tissue‐specific control of EC‐SOD expression: a new mouse model for examining the physiological and pathological roles of extracellular superoxide
Author(s) -
Zou Yani,
Chen Chihhsin,
Huang TingTing
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.823.5
Subject(s) - superoxide dismutase , transgene , microbiology and biotechnology , genetically modified mouse , green fluorescent protein , extracellular , chemistry , biology , oxidative stress , biochemistry , gene
The extracellular isoform of superoxide dismutase (EC‐SOD, Sod3 ) plays a protective role against various injuries mediated by oxidative stress, including acute ischemia, stroke, and hyperoxia. In order to investigate the mechanisms underlining the physiological and pathological roles of EC‐SOD, we designed a Tet‐inducible EC‐SOD construct (TRE‐ Sod3 ‐GFP) which contains a modified TRE promoter (TRE‐Tight), HA‐tagged Sod3 (EC‐SOD/HA), internal ribosome entry site (IRES), and a modified GFP (hrGFP). In vitro expression study showed non‐detectible baseline expression of EC‐SOD, but a robust expression in response to doxycycline induction. Transgenic mice generated from this construct were crossed to transactivator transgenic lines (CamKII‐tTA and LAP‐tTA) to test the expression of the transgene in the forebrain and liver. The results show that TRE‐ Sod3 ‐GFP/CamKII‐tTA double transgenic mice express higher levels of EC‐SOD in hippocampus and cortex, whereas TRE‐ Sod3 ‐GFP/LAP‐tTA have increased EC‐SOD in liver and plasma (as the secreted form). EC‐SOD protein level also correlates well with increased SOD activity in the plasma of TRE‐ Sod3 ‐GFP/LAP‐tTA mice, suggesting that EC‐SOD produced from the transgene are biologically active. The Tet‐inducible TRE‐ Sod3 ‐GFP mouse model will be helpful in interpreting the multiple biological roles of EC‐SOD in different organs. Funding from NIH and PAIRE.