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Calretinin (CR) is a calcium-binding protein that plays an important role in the homeostasis of intracellular calcium concentration in the auditory pathway. To test if hearing loss could lead indirectly to modifications in levels of this calcium-binding protein in neurons and neuropilar structures outside of the lemniscal auditory pathway, CR-immunostaining was evaluated in the superior colliculus (SC) in adult ferrets at 1, 20 and 90 days after unilateral cochlear ablation. The results demonstrate that within 24 h there was a significant increase in CR-immunostaining in ablated animals as indicated by an increase in the mean gray level of immunostaining in the deep, multisensory layers of the contralateral SC compared to the ipsilateral side and control ferrets. This upregulation was evident in both neurons and neuropil and did not change at the two subsequent time points. In contrast, there was no change in the superficial layers of the SC which have visual properties but no auditory inputs. These findings suggest that upregulation of CR levels within neurons and neuropil in the contralateral deep SC is subject to modifications by activity in multisynaptic auditory pathways. Therefore, cochlear-driven activity appears to affect calcium-binding protein levels not only in auditory nuclei but also in other neural structures whose response properties may be influenced by auditory-related activity.