We localized a calcium-binding protein, calretinin, in the superior colliculus of the mouse and studied the distribution and effect of eye enucleation on the distribution of this protein. Calretinin was localized with immunocyto-chemistry. A dense plexus of anti-calretinin-labeled fibers was found within the superficial layers. The highest density was found in the deep superficial gray layer. Monocular enucleation produced an almost complete reduction of calretinin-immunoreactive fibers in the superficial layers of the superior colliculus contralateral to the enucleation. Furthermore, many calretinin-labeled cells appeared in the contralateral superior colliculus. These newly appeared neurons had small oval or round cell bodies. The results demonstrate that calretinin identify unique neuronal sublaminar organizations in the superior colliculus of the mouse. They also suggest that the retinal projection may control in part the content of calretinin in some neurons in the superficial layers of the mouse superior colliculus.