Induction of a 55-kDa PKN cleavage product by ischemia/reperfusion model in the rat retina

作者: Yasuhito Shirai , Kiyoshi Sumioka , Naoaki Saito , Mikiko Takahashi , Yoshitaka Ono

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摘要: PURPOSE. To investigate the physiological role of a protein kinase, PKN, and its relation to apoptosis in vivo. METHODS. An ischemia/reperfusion model rat retina was created by elevating intraocular pressure. Retinal samples were obtained after ischemic insult (15-45 minutes) followed reperfusion (1-7 days). The effect ischemia on fragmentation PKN examined immunoblotting immunocytochemical procedures using antibody against PKN. N-methyl-D-aspartate (NMDA) or caspase-3 inhibitor (DEVD-CHO) administered intravitreally induction fragmentation. retinal cell loss each sample evaluated toluidine blue staining. RESULTS. Ischemia induced 55-kDa cleavage fragment corresponding molecular size constitutively active appearance depended duration correlated with occurrence loss. Immunocytochemical analysis revealed that increased immunoreactivity inner layers retina. DEVD-CHO significantly inhibited protected administration NMDA also CONCLUSIONS. is specifically cleaved related protease during vivo, at least partially initiated activation receptor.

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