Expression and characterization of SPARC in human lens and in the aqueous and vitreous humors.
SPARC (secreted protein, acidic and rich in cysteine) is a matricellular glycoprotein that regulates morphogenesis, cellular proliferation, and differentiation. SPARC is a critical factor in the development and maintenance of lens transparency in mice. SPARC-null mice develop lenticular opacity at an early age that progresses gradually to mature cataract. Despite the high level of homology between the mouse and human genes, little is known about SPARC in the human lens. We have studied the expression of SPARC protein in human lens and surrounding ocular tissues from normal human donors (60-70 years old). Immunohistochemical and immunoblot analyses were conducted on lens, aqueous humor, vitreous, ciliary epithelium, pigment epithelium, cornea and retina. The epithelia and capsule of the lens contained SPARC, whereas the cortical and nuclear fibers did not. In contrast, the aqueous humor and vitreous, which provide nutrients to the lens and regulate its development and function, contained significant amounts of SPARC. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of extracts of various ocular tissues revealed bands of 43 and 29 kD after disulfide bond reduction that were reactive with anti-SPARC IgG. Despite the presence of protease inhibitors during sample preparation, we observed cleavage of intact SPARC to a 29 kD fragment, a peptide reported in other tissues and attributed to endogenous proteolysis. In addition, bands of molecular mass 150 and 200 kD were present that appeared to be disulfide-bonded complexes of SPARC monomers. Human cornea, ciliary epithelium, pigment epithelium and retina also contained SPARC. The presence of SPARC in the aqueous humor and vitreous, as well as in the lens, indicates a functional importance of SPARC in adult human eye as well as in lens development.
Yan
Q
Q
Department of Vascular Biology, The Hope Heart Institute, Seattle, WA 98122, USA.
Clark
J I
JI
Sage
E H
EH
D000368:Aged; D000818:Animals; D001082:Aqueous Humor / Q000378:metabolism*; D002417:Cattle; D002924:Ciliary Body / Q000378:metabolism; D003315:Cornea / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D007908:Lens, Crystalline / Q000378:metabolism*; D008875:Middle Aged; D015676:Osteonectin / Q000378:metabolism*; D010857:Pigment Epithelium of Eye / Q000378:metabolism; D012160:Retina / Q000378:metabolism; D014822:Vitreous Body / Q000378:metabolism*
D015676:Osteonectin