Chondroitin

Composition of perineuronal net extracellular matrix in rat brain: A different disaccharide composition for the net-associated proteoglycans.

Deepa SS, Carulli D, Galtrey C, Rhodes K, Fukuda J, Mikami T, Sugahara K, Fawcett JW.

Centre for Brain Repair, University of Cambridge, Cambridge CB2 2PY.

We developed a method to extract differentially chondroitin sulphate proteoglycans (CSPGs) that are diffusely present in the central nervous system (CNS) matrix and CSPGs that are present in the condensed matrix of perineuronal nets (PNNs). Adult rat brain was sequentially extracted with Tris buffered saline (TBS), TBS containing detergent, 1M NaCl and 6M urea. Extracting tissue sections with these buffers showed that the diffuse and membrane-bound CSPGs were extracted in the first three buffers, but PNN-associated CSPGs remained and were only removed by 6M urea. Most of the CSPGs were extracted to some degree with all the buffers, with neurocan, brevican aggrecan and versican particularly associated with the stable urea-extractable PNNs. The CSPGs in stable complexes only extractable in urea buffer are found from postnatal day 7-14 coinciding with PNN formation. Disaccharide composition analysis indicated a different glycosaminoglycan (GAG) composition for PGs strongly associated with extracellular matrix (ECM). For CS/dermatan sulphate (DS)-GAG the content of nonsulfated, 6-O-sulfated, 2,6-O-disulfated and 4,6-O-disulfated disaccharides were higher and for heparan sulphate (HS)-GAG, the content of 6-O-sulfated, 2-N-, 6-O-disulfated, 2-O-, 2-N-disulfated and 2-O-, 2-N-, 6-O-trisulfated disaccharides were higher in urea extract as compared with other buffer extracts. Digestions with chondroitinase ABC and hyaluronidase indicated that aggrecan, versican, neurocan, brevican and phosphacan are retained in PNNs through binding to hyaluronan (HA). A comparison of the brain and spinal cord ECM with respect to CSPGs indicated that the PNNs in both parts of the CNS have the same composition.

PMID: 16644727 [PubMed - as supplied by publisher]

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