This study describes for the first time the distribution of the calcium-binding protein calretinin (CR) in the anterior thalamic nuclei (ATN) of the guinea pig during development. Brains from animals ranging from 40th embryonic day (E40) to 80th postnatal day (P80) were used in the study. No CR-immunoreactive (CR-ir) perikarya were present among the ATN at E40, but thick bundles of fibers containing CR were crossing the anteromedial nucleus (AM). The first CR-ir neurons appeared at E50 in the lateral part of the AM. At E60, the bundles of fibers disappeared and the whole area of AM displayed closely packed CR-ir perikarya. At this stage, CR also appeared in neurons of the anteroventral nucleus (AV), particularly in its lateral part and along its dorsal border. Moreover, from E50 short and thin bundles of fibers were observed in the medial part of the AV. The ATN of newborns (P0) already showed an adult-like CR distribution pattern - perikarya in the AM and AV were distributed more homogenously and their number was slightly decreased in comparison to E60. The anterodorsal nucleus (AD) was devoid of CR-ir neurons in all studied stages. In conclusion, our results demonstrate that calretinin appears for the first time in neurons of various anterior thalamic nuclei of the guinea pig between 40th and 60th day of prenatal development. (C) 2012 Elsevier B.V. All rights reserved.