In this article, proteoglycans from embryonic chick leg muscle are quantitatively and qualitatively compared with day 8 high density cell culture cartilage proteoglycans by electron microscopy of proteoglycan-cytochrome c monolayers. The visualized proteoglycan profiles were separated into four categories according to shape, size, and complexity. The two major categories were further characterized by lengths of core proteins, lengths of side projections, and distance between side projections. Two large proteoglycans are identifiable in spread leg muscle preparations. One group has a core protein (mean length of 205 nm) from which extend long thin side projections that we interpret to be groups of chondroitin sulfate glycosaminoglycans with a mean length of 79 nm. This large chondroitin sulfate proteoglycan is the only type found in muscle cultures as determined both biochemically in the past and now by electron microscopy and is referred to as muscle proteoglycan. The second large proteoglycan has a mean core protein length of 250 nm and side projections that are visibly shorter (mean length of 38 nm) and thicker than those of the muscle proteoglycan. This group is referred to as the mesenchymal proteoglycan since its biosynthetic origin is still uncertain. We compare these two profiles with the chick cartilage chondroitin sulfate proteoglycan that has a mean core protein length of 202 nm and side projections with a mean length of 50 nm. The data presented here substantiate the earlier biochemical characterization of these noncartilage proteoglycans and establish the unique structural features of the muscle proteoglycan as compared with the similar profiles of the cartilage and mesenchymal proteoglycans.The Kleinschmidt cytochrome c monolayer spreading technique (20, 32, 33) has been used to visualize and characterize proteoglycans. These studies have concentrated on cartilage (2, 4-6, 25, 28, 32-34) or cartilage tumor proteoglycans (3, 17) and have reported on the size and complexity of aggregates (3,4,6,17,32,33), changes in proteoglycans with age or anatomical location (2, 5, 25, 28), and differences between organisms (4). In this context, this laboratory has reported details of transmission electron microscope studies and biochemical characterization of day 8 high density chick limb bud culture chondrocyte proteoglycans (28). The day 8 chick limb bud chondrocyte proteoglycan has a molecular weight of-2 x 106 with chondroitin sulfate chains of ~20,000 mol wt, which contain 67% 6-sulfated chondroitin sulfate. Although cartilage and chondrosarcoma proteoglycans have received substantial attention, proteoglycans from several tissue sources have been characterized biochemically (1,11,12,14,15,18,19,22,23,27, 29, 31, 35-37).We recently biochemicaUy characterized newly synthesized chick cartilage and muscle proteoglycans isolated from embryos (12, 14) as a means of comparing their structure with that of the proteoglycans synthesized in cell cultures (11). In the study of in vivo proteoglycans synthesized ...