The ontogenetic development of afferent (supraspinal and propriospinal) as well as efferent (ascending) fiber connections of the spinal cord was examined following the injection of horseradish peroxidase (HRP) or wheat germ agglutinin HRP (WGA-HRP) into the cervical and lumbar spinal cords (or brains) of embryos ranging in age from 4 to 14 days of incubation. A few cells were first reliably retrogradely labelled in the pontine reticular formation on embryonic day (E) 4 and E5 following the injection of WGA-HRP into the cervical and lumbar spinal cord, respectively. Propriospinal projections to the lumbar spinal cord, originating from brachial spinal cord, were found by E5, and from the cervical spinal cord by E5.5. Ascending fibers arising from neurons in the lumbar spinal cord could be followed to rostral mesencephalic levels in E5 embryos. Thus, the earliest supraspinal, propriospinal, and ascending fiber connections appear to be formed almost simultaneously. Retrogradely labelled cells were found in the raphe, reticular, vestibular, interstitial, and hypothalamic nuclei in E5.5 embryos following lumbar injections of WGA-HRP. Except for neurons in cerebellar nuclei, all the cell groups of origin that project to the cervical spinal cord of posthatching chicks were also retrogradely labelled by E8. There was a delay in the time of appearance of the projections from various regions of the brain stem to the lumbar versus the cervical spinal cord, ranging from 0.5 to 7 days, but typically of about 3 days duration. A large number of cells located in the ventral hypothalamic region, just dorsal to the optic chiasma, were found to be labelled following cervical HRP injection between E6 and E10. These cells may represent transient projections that are present only during embryonic stages since no labelled cells were found in this region in the newly-hatched chick.