[Study of bcl-X(L) protection on retinal photoreceptors from apoptosis in vitro].
OBJECTIVE
To assess the effect of bcl-X(L), an anti-apoptotic gene, on glutamate-induced apoptosis in cultured retinal photoreceptors.


METHODS
Glutamate-induced apoptosis in cultured retinal photoreceptors was established. The experiment was divided into three groups: control, glutamate treatment and rAd-gfp-bcl-X(L) + glutamate transfection group, the protection of bcl-X(L) on retinal photoreceptors from apoptosis was evaluated. 6.5 x 10(12) pfu/L rAd-gfp-bcl-X(L) were transfected into retinal photoreceptors in the rAd-gfp-bcl-X(L) + glutamate group 48 h before Glutamate-induced apoptosis was established. The positive photoreceptors with green fluorescence were identified under fluorescence microscopy. The expression of Bcl-X(2) protein in rAd-gfp-bcl-X(L) transfected and non-transfected neurons were assessed by immunohistochemistry assay. DNA fragment was detected by agarose gel electrophoresis. Nuclei were revealed by hoechst33258 staining.


RESULTS
rAd-mediated gene delivery can transfect retinal photoreceptors effectively. Increased expression of Bcl-X(L) protein was demonstrated in the rAd-transfecting neurons. Transfection of bcl-X(L) significantly decreased the number of apoptotic retinal photoreceptors in vitro.


CONCLUSIONS
Transfection of bcl-X(L) protects cultured retinal photoreceptors from apoptosis induced by glutamate. Transfection the gene of bcl-X(L) may be a potential gene therapy for retinal degenerative diseases.
Tang
Shi-bo
SB
Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China. sbtang@gzsums.edu.cn
Luo
Yan
Y
Yang
Bin
B
Lin
Shao-fen
SF
Lin
Jian-xian
JX
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D066298:In Vitro Techniques; D010786:Photoreceptor Cells / Q000187:drug effects* / Q000473:pathology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D051020:bcl-X Protein / Q000235:genetics / Q000494:pharmacology*
D051020:bcl-X Protein