It has been suggested that an extracellular matrix - and cell surface - associated glycoprotein, fibronectin, plays a role in the positioning of cells in morphogenesis and in the maintenance of orderly tissue organization. In the present study the appearance and distribution of fibronectin during in ovo chick limb development has been investigated by indirect immunofluorescence techniques in H.H. stages 20–30. Fibronectin is not detectable until just prior to the transition from the morphogenetic to the cytodifferentiation phase of development. Beginning at H.H. stage 25, successive nonrandom patterns of fibronectin detection and distribution, which resemble the subsequent cartilaginous elements, precede overt chondrogenesis as detected by Alcian blue staining. This corresponds to the onset of the cytodifferentiation phase of limb development. As the accumulation of acidic proteoglycan increases in the cartilage matrix and the mesenchymal cells become more round in appearance, the presence of detectable fibronectin decreases and is ultimately seen only in the perichondria and basement membrane. However, predigestion of developed cartilage tissue with testicular hyaluronidase, prior to fibronectin staining, indicated that fibronectin remains a major constituent of cartilage matrix and is apparently masked by cartilagespecific proteoglycans. This study of chick limb development is consistent with the hypothesis that fibronectin may be a molecule that facilitates the spatial organization of cartilaginous primordia cytodifferentiation.

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