Isolation and immunohistochemical localization of a chondroitin sulfate proteoglycan from adult rat brain.
A chondroitin sulfate proteoglycan called PGM1 has been isolated from the particulate fraction of adult rat forebrain. Delipidation of the material, solubilization of proteoglycans in guanidinium chloride, precipitation at low ionic strength, and final extraction at pH 5.0 were used for its isolation. Proteoglycans were subjected to further purification by diethylaminoethyl-cellulose chromatography. Individual components were separated by gel filtration. PGM1 appeared to be a high-molecular-weight chondroitin sulfate proteoglycan, capable of strong interaction with hyaluronic acid. It was finally isolated by gel filtration on Ultrogel AcA 22 in the presence of 4 M guanidinium chloride. Monospecific antibodies obtained in rabbits against the purified molecule did not cross-react with other brain proteoglycans. Immunocytochemical techniques revealed an almost unique association of this compound with axons, particularly those known to contain neurofilaments. However, not all these axons and all parts of these axons contained PGM1. This component was not detectable in liver, intestine, spleen, kidney, lung, heart, skin, hair, lens, and muscle, a finding suggesting a specificity for the nervous tissue. This component is expressed in neural cell cultures. Despite the preservation of the neuronal specificity, it seems to lose its specific axonal localization in vitro.
Normand
G
G
Centre de Neurochimie du CNRS, Strasbourg, France.
Kuchler
S
S
Meyer
A
A
Vincendon
G
G
Zanetta
J P
JP
D000818:Animals; D000918:Antibody Specificity; D001369:Axons / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D011508:Chondroitin Sulfate Proteoglycans / Q000302:isolation & purification* / Q000378:metabolism; D002810:Chondroitinases and Chondroitin Lyases; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006821:Hyaluronoglucosaminidase; D007150:Immunohistochemistry; D009413:Nerve Fibers, Myelinated / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D011509:Proteoglycans / Q000032:analysis / Q000302:isolation & purification*; D051381:Rats; D011919:Rats, Inbred Strains; D014018:Tissue Distribution
D011508:Chondroitin Sulfate Proteoglycans; D011509:Proteoglycans; D006821:Hyaluronoglucosaminidase; D002810:Chondroitinases and Chondroitin Lyases