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An autoradiographic and electron microscopic study of collagen synthesis in differentiating cartilage

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Summary

The synthesis of the proline-rich collagen component of cartilage matrix has been studied by autoradiography using both the light and electron microscope. Amblystoma maculatum larvae had their forelimbs amputated, were allowed to regenerate for 12–15 days, and then injected intraperitoneally with tritiated proline. The animals were fixed at various times (1 min. to 28 days) after the injection and sections of the developing limbs were coated for autoradiography by dipping in Ilford L 4 or Gevaert 3.07 emulsion. The sequential labeling of the organelles of the cartilage cell which occurred is illustrated in light and electron micrographs. Radioactive products first appeared in the ergastoplasm and were associated with the cisternae of the endoplasmic reticulum. Twenty to thirty minutes after the injection, labeled material began to appear in the Golgi zone. There, the newly synthesized protein accumulated within large vacuoles. The fibrillar material within the vacuoles may represent collagen and the more amorphous material, mucoprotein. The vacuoles subsequently (∼2 hrs. later) discharge their labeled contents into the extracellular space. The secreted protein is probably soluble collagen (tropocollagen) for it diffuses readily through the matrix to polymerize into striated collagen fibrils some distance from the cell. These findings contradict some widely held opinions that the fibrillar component of the matrix arises by excortication and appositional growth of fibrils originating from the ectoplasm of chondrocytes. It seems reasonable to conclude that the secretory pathway by which extracellular proteins are produced in cartilage is analogous to that suggested for epithelial gland cells.

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  1. Department of Anatomy, Harvard Medical School, Boston 15, Massachusetts, USA

    Jean -Paul Revel & Elizabeth D. Hay

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Supported by grants CA 05196-04S1 and GM-K3-13, 979-C1-A from the United States Public Health Service.

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Revel, J.P., Hay, E.D. An autoradiographic and electron microscopic study of collagen synthesis in differentiating cartilage. Zeitschrift für Zellforschung 61, 110–144 (1963). https://doi.org/10.1007/BF00341524

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