Chondrogenesis was monitored in micromass cultures of mesenchymal cells derived from the distal tip of stage-25 chick limb buds over a 6-day period. Alcian green staining and immunofluorescent localization of cartilage-specific proteoglycans revealed the appearance of cartilage matrix by day 3 of cell culture. By day 6, cultures contained a uniform and homogeneous population of fully differentiated chondrocytes throughout the cell layer, with only a narrow rim of nonchondrogenic cells around the extreme periphery of the culture. Synthesis of sulfated glycosaminoglycans also progressively increased between days 3 and 6, being 8-fold higher at day 6 than at day 1 of culture. Both adenylate cyclase (AC) activity and cAMP concentrations increased dramatically during the first 2 days of culture, reaching maximal levels by day 2, which remained elevated and stable throughout the remaining chondrogenic period (days 3-6). Responsiveness of both AC and cAMP concentrations of the cells to PGE2 was maximal by day 1 of culture and was increased over control cells by 12-fold and 8-fold respectively. Both responses, however, were dramatically reduced by day 3, at which time the initiation of cartilage formation was apparent. Responsiveness of cells during the prechondrogenic period to PGE2 was relatively specific in that no effects could be demonstrated with equivalent concentrations of PGF2 alpha or 6-keto-PGF1 alpha, although PGl2 did produce increases in cAMP concentrations of about 50% of those of PGE2. These results indicate that previously reported changes in the cAMP system in heterogeneous cell cultures derived from whole limb buds reflect changes occurring in the chondrogenic cell type and indicate further that peak responsiveness of the cAMP system of these cells to prostaglandins is restricted to prechondrogenic developmental periods.
Light cells in the hamster thyroid gland were examined by electron microscopy and histochemical methods in control, hypercalcemic and hypocalcemic animals. Elevation of plasma calcium levels stimulated a marked depletion of light cell granules as detected by electron microscopy and silver impregnation. In the hypercalcemic state the size and number of light cells did not differ from that of control animals. Hypocalcemic hamsters showed a significant increase in light cell numbers with marked hypertrophy and hyperplasia of this cell type. These observations support the view that the light cell in the hamster thyroid is associated with thyrocalcitonin secretion.
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