Proteoglycans synthesized by embryonic chicken retina in culture: composition and compartmentalization

Characteristics of the chondroitin sulfate/dermatan sulfate proteoglycans (CS/DSPGs) and heparan sulfate proteoglycans (HSPGs) from retinas of 14-day chicken embryos were examined following specific lyase digestion of the HSPG and CS/DSPG glycosaminoglycans, respectively. On the basis of gel exclusion chromatography the prevalent CS/DSPGs in the tissue were above Mr 400 X 10(3) with two or three glycosaminoglycan chains of Mr 60-70 X 10(3). The HSPGs existed in two distinct populations in the tissue. Those in the dominant population appeared to be in the range of Mr 250-300 X 10(3) with 9 to 12 glycosaminoglycan chains of Mr 15-25 X 10(3). The other population consisted of free heparan sulfate chains of Mr 15-25 X 10(3). The HSPGs in the medium tended to be intermediate in size. To examine the distribution of proteoglycans, tissues were sequentially homogenized and extracted in saline and reextracted with 4 M guanidine HCl (GdnHCl) and Triton X-100 (TX), or they were washed in heparin solution and dissociated to single cells with trypsin before sequential extraction in saline and GdnHCl with TX. Through comparison of the results of these two extraction methods, CS/DSPGs were found to be almost entirely within the medium or matrix or loosely associated with the cell surface, and most HSPGs were associated with either the basal lamina or the plasma membrane. The single heparan sulfate glycosaminoglycan chains appeared to be intracellular degradation products. These results support reports that CS/DSPGs may be present in the retina interphotoreceptor matrix and that HSPGs may be present in regions of synaptogenesis, associated with cell membranes.