Chondroitin sulfate proteoglycan core proteins in the interphotoreceptor matrix: a comparative study using biochemical and immunohistochemical analysis.
This study characterizes the core proteins of chondroitin sulfate-type glycosaminoglycans located in the interphotoreceptor matrix and establishes the tissue distribution of chondroitin immunoreactivity in human, bovine, mouse and rat retinas. Monoclonal antibodies specific to unsulfated (DeltaDiOS), 4-sulfated (DeltaDi4S) and 6-sulfated (DeltaDi6S) chondroitin were employed. Retinal sections and IPM samples were either (a) digested with chondroitinase ABC to expose antibody specific epitopes, (b) double digested with chondroitinase ABC and chondroitinase AC II to remove specific epitopes, or (c) left undigested to evaluate mimotope labeling. In tissue sections from each species studied, positive immunoreactivity to the DeltaDi6S antibody was present in the IPM surrounding both rods and cones. In human and bovine, DeltaDi6S labeling of the cone matrix compartments was more intense than labeling of the matrix surrounding rods. Intense DeltaDi6S immunoreactivity was present surrounding the foveal cones. In mouse and rat, no differences in labeling intensity of IPM surrounding rod and cone photoreceptors were evident, although labeling of the IPM near the apical surface of the retinal pigment epithelium and around the photoreceptor inner segments was more pronounced than that surrounding the outer segments. All DeltaDi6S antibody labeling was eliminated with chondroitinase AC II digestion. No IPM immunoreactivity in tissue sections was observed when the DeltaDi0S or DeltaDi4S antibodies were used. In Western blots of IPM extracts treated with chondroitinase ABC, prominent DeltaDi6S immunoreactive bands were present at approximately 230 kD and 150 kD in each species studied, with the exception of the human, where the 150 kD component is not a chondroitin proteoglycan. Each of the prominent DeltaDi6S immunoreactive bands showed minor immunoreactivity to the DeltaDi4S antibody. No DeltaDi0S immunoreactivity was noted in Western blots of IPM samples from any species. All immunoreactivity was lost following chondroitinase AC II digestion. These observations document similarities in the electrophoretic mobility of IPM proteoglycan core proteins released following chondroitinase ABC digestion in the four species studied, but reveal pronounced differences in the tissue distribution. Bovine and human IPM show greater concentrations of DeltaDi6S immunoreactivity surrounding cones than rods, whereas rodent tissues show higher concentrations near the retinal pigment epithelium and around the photoreceptor inner segments than around the outer segments. The pattern of distribution of these proteoglycan molecules is highly conserved in these species, suggesting a common role in IPM structure and function.
Hollyfield
J G
JG
The Cole Eye Institute, The Cleveland Clinic Foundation, Cleveland, OH, 44195, USA.
Rayborn
M E
ME
Midura
R J
RJ
Shadrach
K G
KG
Acharya
S
S
D000328:Adult; D000368:Aged; D000818:Animals; D002417:Cattle / Q000378:metabolism; D002809:Chondroitin Sulfates / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D005136:Eye Proteins / Q000378:metabolism*; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008322:Mammals / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C / Q000378:metabolism; D008875:Middle Aged; D008969:Molecular Sequence Data; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism*; D011509:Proteoglycans / Q000378:metabolism*; D051381:Rats / Q000378:metabolism; D013045:Species Specificity
D005136:Eye Proteins; D011509:Proteoglycans; D002809:Chondroitin Sulfates