Interphotoreceptor matrix in the fovea and peripheral retina of the primate Macaca mulatta: distribution and glycoforms of SPACR and SPACRCAN.
SPACR and SPACRCAN localization in the interphotoreceptor matrix (IPM) of the fovea and peripheral retina of Macaca mulatta was established with antibodies to these core proteins and the chondroitin sulfate epitopes and lectin binding properties of these molecules were defined. The IPM of both rods and cones labeled with anti-SPACR, anti-SPACRCAN, anti-Delta Di6S antibodies and wheat germ agglutinin (WGA). Whereas anti-SPACR and anti-SPACRCAN antibodies labeled rod and cone matrix compartments with similar intensity, the Delta Di6S chondroitin antibody labeling was more intense around cones than rods. Peanut lectin (PNA) labeling was present only around cones. No IPM labeling was observed with Delta Di0S-chondroitin or Delta Di4S-chondroitin antibodies. Western blots of undigested IPM extracts showed anti-SPACR immunoreactivity at 150 kDa, colocalizing with the position of WGA and PNA binding. In Western blots of the chondroitinase ABC digested sample and samples double digested with chondroitinase ABC and AC II, anti-SPACR immunoreactivity, WGA and PNA labeling intensity were virtually identical to that in the undigested sample, with prominent staining of the 150 kDa SPACR band. In contrast, anti-SPACRCAN immunoreactivity was not present in the undigested sample, but was evident in both the chondroitinase ABC and double digested samples as a broad band at approximately 230 kDa. Delta Di6S, Delta Di4S, WGA and PNA labeling colocalized with the anti-SPACRCAN immunoreactivity in the chondroitinase ABC digested sample. These findings indicate that SPACR and SPACRCAN are present around cones in the fovea and both rods and cones in the peripheral retina, but that the specific glycoforms of these molecules are different depending on whether present in the cone or rod associated IPM.
Hollyfield
J G
JG
Cole Eye Institute, The Cleveland Clinic Foundation, Cleveland, OH 44195, USA. hollyfj@ccf.org
Rayborn
M E
ME
Nishiyama
K
K
Shadrach
K G
KG
Miyagi
M
M
Crabb
J W
JW
Rodriguez
I R
IR
D000818:Animals; D015153:Blotting, Western; D019765:Chondroitin ABC Lyase; D004591:Electrophoresis, Polyacrylamide Gel; D008253:Macaca mulatta; D013058:Mass Spectrometry; D015676:Osteonectin / Q000032:analysis* / Q000378:metabolism; D010449:Peptide Mapping; D020419:Photoreceptor Cells, Vertebrate / Q000737:chemistry*
D015676:Osteonectin; D019765:Chondroitin ABC Lyase