The expression pattern of the extracellular matrix molecule tenascin‐C was investigated in the retinotectal system of the frog Discoglossus pictus and the salamander Pleurodeles waltl during development and optic nerve regeneration in the adult. In both species, the retina was devoid of tenascin‐C immunoreactivity at all ages studied. During development, tenascin‐C was distributed in a gradient in the optic nerve, with the highest immunoreactivity in the eye near part of the optic nerve. The myelin‐associated glycoprotein was distributed in a gradient with opposite polarity. In Discoglossus, but not Pleurodeles, tenascin‐C was detected in the anterior chiasm. In the tectum of both species, tenascin‐C was observed in deep cellular and fiber layers but not in the layers receiving optic fibers or proliferative zones. The distribution patterns of tenascin‐C were the same during development and in the adult, except for a disappearance of the molecule from the intraocular part of the optic nerve. After lesioning the optic nerve of adult animals, tenascin‐C was strongly reexpressed in the intraocular part of the optic nerve but was only weakly upregulated in the distal optic nerve stump. In contrast, a chondroitin sulfate epitope was strongly upregulated in the distal optic nerve stump. These observations suggest that during development, tenascin‐C serves as an attenuating barrier for myelinating cells in the optic nerve and contributes to the guidance of growing retinal ganglion cell axons. Due to its sustained expression in the adult, tenascin‐C may have similar functions during regeneration of the lesioned adult retinotectal system. © 1995 Wiley‐Liss, Inc.