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1 Ophthalmology, Wakayama Medical Univeersity, Wakayama, Japan
2 Pathology, Wakayama Medical University, Wakayama, Japan
3 Anatomy, , Graduate School of Medicine, Osaka City University, Osaka, Japan
4 Laboratory of Cell Regulation and Carcinogenesis, NCI/NIH, Bethesda, USA
* To whom correspondence should be addressed. E-mail: shizuya{at}wakayama-med.ac.jp.
We examined the effect of adenovirus-mediated expression of bone morphogenic protein-7 (BMP-7) and inhibitors of differentiation 2 and 3 (Id2 and Id3) on injury-induced epithelial-mesenchymal transition (EMT) of lens epithelium in mice. Id2 and Id3 are known to be up-regulated by BMP-7 and to antagonize Smad2/3 signaling. The Cre-LoxP system adenoviral gene transfer was employed. Three µl of adenoviral solution (2 x 107 PFU/µl) was injected into the right lens of adult male C57BL/6 mice (n = 144) at the time of capsular injury made using a hypodermic needle under both general and topical anesthesia. A mixture of Cre-Ad and vector encoding mBMP-7, mId2 or mId3 was administered in test group. Control lenses were treated with Cre-Ad alone. After healing intervals of 5 or 10 day, the animals were killed and processed histology or RNA extraction from lens. Reverse transcription polymerase chain reaction (RT-PCR), real-time RT-PCR and immunohistochemistry showed expression of each introduced gene in the lens. Exogenous BMP-7 up-regulated expression of Id2 and Id3 in injured lenses and gene introduction of Id2 or Id3 also up-regulated BMP-7 expression. Gene transfer of BMP-7, Id2 or Id3 delayed injury-induced EMT of the lens epithelial cells as evaluated by histology and expression patterns of 
-smooth muscle actin and collagens in association with reduction of Smad2 C-terminal phosphorylation. Gene transfer of BMP-7, Id2 or Id3 delayed injury-induced EMT of lens epithelial cells and subsequent sealing of the capsular break with fibrous tissue in mice.
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