Chondroitin sulfate proteoglycan is a constituent of the basement membrane in the rat embryo parietal yolk sac.
In addition to containing Type IV collagen, laminin and entactin, basement membranes contain small amounts of proteoglycans substituted primarily with heparan sulfate chains. We have previously shown, however, that parietal yolk sacs in organ culture synthesize predominantly chondroitin sulfate proteoglycan. In the present study, we have used histochemical and immunohistochemical techniques coupled with chondroitinase ABC digestion to provide evidence for the presence of chondroitin sulfate proteoglycan in the basement membrane (Reichert's membrane) of the 14.5-day rat embryo parietal yolk sac. The results revealed numerous cuprolinic blue-positive filaments and granules, 20-30 nm in greater length or diameter, dispersed throughout the thickness of the basement membrane. Both structures were removed by preincubating freshly isolated parietal yolk sacs with chondroitinase ABC. A similar labeling pattern was also obtained with immunoelectron microscopy using gold-labeled monoclonal antibodies directed against the three major isomers of protein-bound chondroitin sulfate. In contrast, coarser cuprolinic blue granules, 40-100 nm in diameter, were neither sensitive to chondroitinase ABC digestion nor labeled by the monoclonal antibodies. These results thus indicate that Reichert's membrane contains chondroitin sulfate proteoglycan in addition to heparan sulfate proteoglycan.
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104.
R V
RV
Iozzo
C C
CC
Clark
D000818:Animals; D001485:Basement Membrane / Q000032:analysis; D002808:Chondroitin Lyases / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis*; D004396:Coloring Agents; D006651:Histocytochemistry; D007150:Immunohistochemistry; D007211:Indoles; D008854:Microscopy, Electron; D009942:Organometallic Compounds; D011509:Proteoglycans / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D015017:Yolk Sac / Q000032:analysis*
D011508:Chondroitin Sulfate Proteoglycans; D004396:Coloring Agents; D007211:Indoles; D009942:Organometallic Compounds; D011509:Proteoglycans; C015445:copper phthalocyanine; D002808:Chondroitin Lyases