Relation between the time-spatiotemporal expression of caspase-9 and photoreceptor apoptosis as well as electroretinography in RCS rats

AIM: To investigate the relation between the time- spatiotemporal expression of caspase-9 and photoreceptor apoptosis as well as electroretinography (ERG) in Royal College of Surgeons (RCS) rats. METHODS: Twelve non-pigmented degenerative RCS rats at age of 15, 20, 25, 30 days were sacrificed after ERG examination. The caspase-9 expression was analyzed with immunohistochemical staining, fluoro-metric activity assay and Western blotting. Apoptotic retinal neurons were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique. RESULTS: The latency and amplitude of b-wave were (58.60±3.42) ms and (109.07±14.50) mV, (56.45±1.08) ms and (109.00±7.35) mV, respectively, in 15 and 20 days old degenerative RCS rats. The ERG resembled flat form in the degenerative RCS rats at age of 25 and 30 days. TUNEL-positive cells were located only in the outer nuclear layer, mostly prevailed in 25-day-old group. Caspase-9 protein expressed in the rat retina started to be found immunohistochemically in 25-day-old and was most predominant in the inner segment. Fluorometric activity assay showed that caspase-9 activity reached its peak, (10.98±0.13) nmol·g-1·min-1 at 20th day. Western blot assay revealed the small cleavage fragmentation at 20th day. No cleavage fragmentation was revealed in the 15-day-old degenerative RCS rats and all other control rats. CONCLUSION: The activation and time-spatiotemporal expression of caspase-9 are coincidence with photoreceptor apoptosis as well as the loss of vision function in RCS degenerative rats, which implicates the important role of caspase-9 in photoreceptor apoptosis.