title: Isolation, characterization, and localization of glycosaminoglycans in rabbit bone marrow.
abstract: The glycosaminoglycans that exist in rabbit bone marrow were analyzed chemically, and their in situ localization was studied immunohistochemically. Femoral bone marrow of 3-month-old rabbits was defatted with organic solvents. Glycosaminoglycans were prepared from the defatted tissue after its digestion with pronase, treatment with mild alkali, and then digestion with DNase-I. The tissue contained glycosaminoglycans equivalent to 195 mg of hexosamine per femur, which accounted for 27.3% of the total hexosamine in the tissue. Studies with hyaluronidase from Streptomyces hyalurolyticus and chondroitinase ABC showed that the glycosaminoglycans were composed of hyaluronic acid (16% of the total glycosaminoglycan) and chondroitin 6-sulfate (79%). The chondroitin 6-sulfate was separated on Bio-Gel A-0.5m gel into two molecular species with mol wt of greater than 12,000 (Kd greater than 0.2) and approximately 8,000 (Kd = 0.47). Bone marrow digested with chondroitinase ABC and then treated with three monoclonal antibodies 4/8/9-A-2, 5/6/3-B-3, and 5/6/1-B-5, which were specific for unsaturated 4-sulfated, 6-sulfated, and nonsulfated disaccharide structures, respectively, at the nonreducing end of chondroitin sulfate chains, reacted with only 5/6/3-B-3. This result indicated that the chondroitin sulfate, isomer in the bone marrow is chondroitin 6-sulfate, consistent with the biochemical results. The chondroitin 6-sulfate was localized mainly in the extracellular compartment and was considered to be involved in construction of the hemopoietic microenvironment in the bone marrow.
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D001853:Bone Marrow / Q000378:metabolism*; D055598:Chemical Phenomena; D002621:Chemistry; D002809:Chondroitin Sulfates / Q000378:metabolism; D005260:Female; D006025:Glycosaminoglycans / Q000302:isolation & purification* / Q000378:metabolism; D007536:Isomerism; D011817:Rabbits; D014018:Tissue Distribution
chemical_list: D000911:Antibodies, Monoclonal; D006025:Glycosaminoglycans; D002809:Chondroitin Sulfates