Isolation, characterization, and localization of glycosaminoglycans in
rabbit bone marrow
K Oguri, E Okayama, B Caterson and M Okayama
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.
Volume 70,
Issue 2,
pp. 501-510,
08/01/1987
Copyright © 1987 by The American Society of Hematology
