Increased expression of osteonectin/SPARC mRNA and protein in age-related human cataracts and spatial expression in the normal human lens

PURPOSE: We have previously reported increased levels of Osteonectin/SPARC transcript in age-related cataractous compared to normal human lenses. The purpose of the present study was to evaluate the corresponding levels of os teonectin/SPARC protein in age-related cataractous relative to normal lense s and to evaluate the levels of osteonectin/SPARC transcript in specific ty pes of age-related human cataracts. The spatial expression of osteonectin/S PARC was also evaluated in normal human lenses. METHODS: Specific types of age-related cataracts were collected and graded. Normal human lenses were microdissected into epithelia and fibers. Osteone ctin/SPARC protein levels were monitored by Western immunoblotting, and tra nscript levels were evaluated by reverse transcriptase polymerase chain rea ction (RT-PCR). Osteonectin/SPARC expression patterns were examined by RT-P CR and by immunostaining. RESULTS: Higher levels of osteonectin/SPARC protein were detected in age-re lated cataractous relative to normal human lenses. Increased levels of oste onectin/SPARC transcript were also detected in posterior-subcapsular and nu clear cataractous lenses relative to normal lenses. Osteonectin/SPARC trans cripts were detected in the lens epithelium but not fibers. Osteonectin/SPA RC protein levels were highest in the peripheral lens epithelium. CONCLUSIONS: Consistent with our previous studies on osteonectin/SPARC mRNA levels, osteonectin/SPARC protein levels were also elevated in cataractous compared to normal human lenses. Increased levels of osteonectin/SPARC mRN A were also found in nuclear and posterior-subcapsular cataracts relative t o normal lenses. Osteonectin/SPARC expression is confined to the lens epith elium, and osteonectin/SPARC levels are highest in the peripheral lens epit helium.