The pattern of glutamate-like immunoreactivity was investigated in the pigeon optic tectum. The most impressive aspect of the labelling pattern was an accumulation of immunoreactive terminal-like elements restricted to those superficial tectal layers that correspond to the termination zone of the retinal afferents. These immunoreactive puncta occurred frequently in small clusters. At the level of electron microscopy, many of the labelled nerve endings showed the characteristics of retinal terminals. Moreover, following unilateral retinal ablation a drastic loss of immunoreactive terminal-like puncta was observed in the retinorecipient layers of the tectum contralateral to the lesion. The remaining glutamate-immunoreactive terminal-like elements had the light and electron microscopic features typical of the afferents from the nucleus isthmi, pars parvocellularis (lpc). The relation between the latter result and the transmitter specificity of the afferents from this subtectal nucleus is unclear at present. On the other hand, the light and electron microscopic labelling patterns and the effect of retinal ablation suggest that afferents from retina and from lpc are the only major sources for glutamate-immunoreactive terminals in the pigeon optic tectum. Furthermore, the results are well in line with previous data indicating glutamate as neurotransmitter at least in part of the retinal afferents to the pigeon optic tectum.