PURPOSE: To compare the abundance and spatial distribution of serum albumin in the mouse and bovine cornea. METHODS: Serum albumin from cornea was separated from transketolase by SDS-PAGE (+/-dithiothreitol [DTT]) and identified by peptide sequencing and immunoblot analyses. The fractional content of serum albumin was determined in water-soluble extracts of cornea by imaging analyses after SDS-PAGE. Serum albumin was localized in cornea by immunohistochemistry and by SDS-PAGE analyses of samples from separated epithelium and stroma. RESULTS: SDS-PAGE (-DTT) resolved mouse serum albumin and transketolase and indicated that serum albumin was 13% of the water-soluble protein in whole mouse corneas. By contrast, corneal epithelial fractions contained little (<1%) serum albumin. Immunohistochemistry indicated that mouse serum albumin was present throughout the stroma between collagen lamellae. Immunohistochemical analyses of bovine cornea yielded similar results. In addition, immunohistochemistry for serum albumin revealed positive staining in a small number of basal epithelial cells next to Bowman's membrane, and greater staining in the anterior-peripheral stroma as well as immediately adjacent to Descemet's membrane. CONCLUSIONS: Mouse and bovine cornea have a similar content and spatial distribution of serum albumin. The appreciable serum albumin in the cornea documented here and elsewhere raise the possibility that it contributes to the physiological or optical functions of the cornea. Moreover, serum albumin's ability to bind drugs suggests that mice corneas could be exploited to study drug-serum albumin interactions in vivo and to test the usefulness of serum albumin as a drug carrier for corneal disorders.
PURPOSE: To compare the abundance and spatial distribution of serum albumin in the mouse and bovine cornea. METHODS: Serum albumin from cornea was separated from transketolase by SDS-PAGE (+/-dithiothreitol [DTT]) and identified by peptide sequencing and immunoblot analyses. The fractional content of serum albumin was determined in water-soluble extracts of cornea by imaging analyses after SDS-PAGE. Serum albumin was localized in cornea by immunohistochemistry and by SDS-PAGE analyses of samples from separated epithelium and stroma. RESULTS:SDS-PAGE (-DTT) resolved mouse serum albumin and transketolase and indicated that serum albumin was 13% of the water-soluble protein in whole mouse corneas. By contrast, corneal epithelial fractions contained little (<1%) serum albumin. Immunohistochemistry indicated that mouse serum albumin was present throughout the stroma between collagen lamellae. Immunohistochemical analyses of bovine cornea yielded similar results. In addition, immunohistochemistry for serum albumin revealed positive staining in a small number of basal epithelial cells next to Bowman's membrane, and greater staining in the anterior-peripheral stroma as well as immediately adjacent to Descemet's membrane. CONCLUSIONS:Mouse and bovine cornea have a similar content and spatial distribution of serum albumin. The appreciable serum albumin in the cornea documented here and elsewhere raise the possibility that it contributes to the physiological or optical functions of the cornea. Moreover, serum albumin's ability to bind drugs suggests that mice corneas could be exploited to study drug-serum albumin interactions in vivo and to test the usefulness of serum albumin as a drug carrier for corneal disorders.
Authors: Natalie Lassen; J Bronwyn Bateman; Tia Estey; Jer R Kuszak; David W Nees; Joram Piatigorsky; Gregg Duester; Brian J Day; Jie Huang; Lisa M Hines; Vasilis Vasiliou Journal: J Biol Chem Date: 2007-06-13 Impact factor: 5.157