Abstract
The accumulation of chondroitin sulfate proteoglycan was correlated with chick limb bud chondrogenesis by immunohistochemical methods. Antisera directed against chondroitin sulfate proteoglycan subunit were used in indirect immunohistochemical reactions which also utilized goat anti-rabbit IgG coupled to fluorescein isothiocyanate, peroxidase, or hemocyanin. Immunohistochemical reaction products were localized to alcian green staining regions in developing limb bud cultures. Developing cartilage nodules reacted with the immunohistochemical probes by 3–4 days and showed a progressive increase in reaction with time. Noncartilage cells in the same cultures did not react. Scanning electron microscopy revealed extensive binding of hemocyanin-antibody complexes to the chondrogenic matrix network. In contrast, the matrix synthesized by limb bud cultures blocked from differentiating by 5-bromo-2′-deoxyuridine showed no reaction.
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