We have shown previously that rubrospinal axons grow around a lesion of their pathway in developing opossums and that a critical period exists for that plasticity. As a first step toward addressing the possibility that glial maturation and/or the development of an astrocytic response to lesioning contribute to loss of rubrospinal plasticity, we have studied the normal development of radial glia and astrocytes in the spinal cord of the opossum by immunostaining for vimentin (Vim) and glial fibrillary acidic protein (GFAP). Vim-like immunoreactivity (Vim-LI) was present in radial glia throughout the spinal cord at birth (12 days after conception), whereas GFAP-like immunoreactivity (GFAP-LI) was limited to cells of comparable morphology in the ventral part of the cervical cord. The subsequent appearance of GFAP-LI followed ventral to dorsal and rostral to caudal gradients and by postnatal day (PD) 15, it was found in radial glia throughout the cord. At the same age, processes immunostained by either antibody had lost their radial orientation in the ventral horn of the cervical cord. The subsequent transformation from radial glia to astrocytes also followed ventral to dorsal and rostral to caudal gradients. By PD30, mature appearing astrocytes were immunostained by both antibodies at thoracic levels of the spinal cord, the levels lesioned in the plasticity experiments referred to above, and by PD41, they were found at all levels of the cord.(ABSTRACT TRUNCATED AT 250 WORDS)