title: Mapping the differential distribution of glycosaminoglycans in the adult human retina, choroid, and sclera.
abstract: PURPOSE. To map the distribution of different classes of glycosaminoglycans (GAGs) in the healthy human retina, choroid, and sclera. METHODS. Frozen tissue sections were made from adult human donor eyes. The GAG chains of proteoglycans (PGs) were detected with antibodies directed against various GAG structures (either directly or after pretreatment with GAG-degrading enzymes); hyaluronan (HA) was detected using biotinylated recombinant G1-domain of human versican. The primary detection reagents were identified with FITC-labeled probes and analyzed by fluorescence microscopy. RESULTS. Heparan sulfate (HS), chondroitin sulfate (CS), dermatan sulfate (DS), and HA were present throughout the retina and choroid, but keratan sulfate (KS) was detected only in the sclera. HS labeling was particularly strong in basement membrane-containing structures, the nerve fiber layer (NFL), and retinal pigment epithelium (RPE)-for example, intense staining was seen with an antibody that binds strongly to sequences containing 3-O-sulfation in the internal limiting membrane (ILM) and in the basement membrane of blood vessels. Unsulfated CS was seen throughout the retina, particularly in the ILM and interphotoreceptor matrix (IPM) with 6-O-sulfated CS also prominent in the IPM. There was labeling for DS throughout the retina and choroid, especially in the NFL, ganglion cell layer, and blood vessels. CONCLUSIONS. The detection of GAG chains with specific probes and fluorescence microscopy provides for the first time a detailed analysis of their compartmentalization in the human retina, by both GAG chain type and sulfation pattern. This reference map provides a basis for understanding the functional regulation of GAG-binding proteins in health and disease processes.
authors: Wellcome Trust Centre for Cell-Matrix Research, UK.
authors: Simon J
authors: SJ
authors: Clark
authors: Tiarnan D L
authors: TD
authors: Keenan
authors: Helen L
authors: HL
authors: Fielder
authors: Lisa J
authors: LJ
authors: Collinson
authors: Rebecca J
authors: RJ
authors: Holley
authors: Catherine L R
authors: CL
authors: Merry
authors: Toin H
authors: TH
authors: van Kuppevelt
authors: Anthony J
authors: AJ
authors: Day
authors: Paul N
authors: PN
authors: Bishop
mesh_terms: D000369:Aged, 80 and over; D002809:Chondroitin Sulfates / Q000378:metabolism; D002829:Choroid / Q000378:metabolism*; D003871:Dermatan Sulfate / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D007632:Keratan Sulfate / Q000378:metabolism; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D012160:Retina / Q000378:metabolism*; D012590:Sclera / Q000378:metabolism*; D014019:Tissue Donors
chemical_list: D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D007632:Keratan Sulfate