Studies on ocular neovascularization by DNA microarray and RNA interference

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Retinal neovascularization in mice retina was produced by the method of Smith et al. The sensory retina was obtained from the model and mRNA expression changes were analyzed by Affimetrix Gene Chip MGU74 AV2. A total of 129 genes were found to be up- or down-regulated by more than twice or half of the control values in the model at P15. Among the up-regulated genes, we paid special attention to βB2-crystallin. βB2-crystallin and a-actin immunoreactivities were detected in glial cells that surrounded neovascular tufts. Short interfering RNA specific to βB2-crystallin were given to the model mice and ocular neovascularization was inhibited by the treatment. mRNA and protein expression were also inhibited by the treatment. βB2-crystallin immunoreactivities were detected in surgically obtained human tissues obtained by vitreous surgery. βB2-crystallin gene was transfected to HEK293 cells and cellular function was analyzed. In transfected cells, elongation of cellular process was observed and βB2-crystallin and a-actin were found to be co-localized underneath the cellular membrane. Our results showed that βB2-crystallin plays an important role in ocular (retinal) neovascularization.