The anatomical distribution of the transcript encoding calcyon, a 24 kDa membrane protein associated with coupling D1-like dopamine receptor activation to potentiated intracellular calcium release, was examined using in situ hybridization in the macaque brain. Calcyon mRNA is found to be abundantly distributed throughout the primate brain. In neocortex, moderately dense, diffuse signal is found in all areas, with increased intensity present in a superficial isodense band corresponding generally to cortical layers II and III. Increased intensity of signal is also seen in the pyramidal cell layers of medial prefrontal and anterior cingulate cortex. Calcyon mRNA is present abundantly in subcortical limbic areas such as the nucleus accumbens, septum, hypothalamus, amygdala, and hippocampus. Moderate calcyon transcript expression is seen in caudate and putamen, with lower levels in globus pallidus. Thalamic nuclei, including the reticular nucleus, express low to moderate levels. Very dense expression is noted in the substantia nigra pars compacta. Numerous brainstem regions express this transcript, notably monoaminergic nuclei including the locus coeruleus and dorsal raphe. The cerebellum has detectable levels of expression in both cortex and deep nuclei. Although calcyon is hypothesized as a means for D1-like receptors to modulate “cross-talk” with other neurotransmitter receptor systems, it is notable that abundant calcyon transcript is detected in brain regions not associated with D1-like neurotransmission, particularly the substantia nigra pars compacta and other dopamine-synthesizing cell groups. A substantial proportion of this may relate to the reported association of calcyon with the D5 receptor, or in addition, may suggest that calcyon has a wider role as a regulator of intracellular signal transduction. J. Comp. Neurol. 468:264–276, 2004. © 2003 Wiley-Liss, Inc.