Qualitative and quantitative analyses of sialic acid in the human trabecular meshwork.
By using colloidal iron and polycationic ferritin at low pH (1.8 and 1.9, respectively) as electron histochemical stains, we localized sialic acid moieties in the trabecular meshwork of normal human eyes. The markers were distributed continuously over the entire surface of the trabecular cells, but were localized linearly as well as randomly in the basal lamina. Within the beams, the markers were located irregularly in the collagen core, but were absent from the elastic tissue and from the 100 nm banded structures in the cortical zone. Pretreatment of the tissue with neuraminidase abolished this staining which indicates that sialated moieties are present in the stained structures. Biochemical analysis with the lectin Limax flavus agglutinin revealed that the major sialated polypeptides in individual samples of human trabecular meshwork migrated at apparent molecular weights of 56, 75, 95, 128, 140, 180 and 220 kDa under reducing conditions. The fractions at approximate molecular weights of 180 and 220 kDa include the sialoglycoproteins thrombospondin and fibronectin, respectively. The total content of sialic acid in the human trabecular meshwork (patient age range, 27-57 yr) was 3.6 +/- 0.6 mumol g-1 wet weight, as determined by a colorimetric assay. We conclude that significant quantities of sialic acid are present in the normal human trabecular meshwork as neuraminidase-sensitive alpha-ketosidically linked terminal residues of the polypeptides.
Tripathi
B J
BJ
Visual Sciences Center, University of Chicago, IL 60637.
Millard
C B
CB
Tripathi
R C
RC
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D003124:Colorimetry; D006651:Histocytochemistry / Q000379:methods; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008854:Microscopy, Electron; D008875:Middle Aged; D019158:N-Acetylneuraminic Acid; D012794:Sialic Acids / Q000032:analysis*; D014129:Trabecular Meshwork / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism
D037102:Lectins; D012794:Sialic Acids; D019158:N-Acetylneuraminic Acid