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Isolation and Immunohistochemical Localization of a Chondroitin Sulfate Proteoglycan from Adult Rat Brain
Author(s) -
Normand Guy,
Kuchler Sabine,
Meyer Alphonse,
Vincendon Guy,
Zanetta JeanPierre
Publication year - 1988
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1988.tb01796.x
Subject(s) - proteoglycan , chondroitin sulfate proteoglycan , guanidinium chloride , chemistry , biochemistry , chondroitin sulfate , neurofilament , chondroitin , size exclusion chromatography , biology , immunohistochemistry , glycosaminoglycan , extracellular matrix , immunology , enzyme
A chondroitin sulfate proteoglycan called PGM 1 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. PGM 1 appeared to be a high‐molecular‐weight chondroitin sulfate proteoglycan, capable of strong interaction with hy‐aluronic 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 PGM 1. 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.