The claustrum is not missing from all monotreme brains

Many authors have reported that the claustrum, which comprises the insular claustrum and the endopiriform nucleus, is missing from the monotreme forebrain. We used Nissl and myelin staining in conjunction with enzyme histochemistry for acetylcholinesterase and immunohistochemistry for parvalbumin, calbindin, calretinin and tyrosine hydroxylase to examine the brains of two monotremes, the short-beaked echidna (Tachyglossus aculeatus) and the platypus (Ornithorhynchus anatinus). We found that although the insular claustrum is a small structure in the echidna brain, it is nevertheless clearly present as loosely clustered neurons embedded in the white matter ventrolateral to the putamen and deep to the piriform and entorhinal cortices. Neurons in this region share the chemical features of the adjacent cortex (presence of a similar proportion of parvalbumin immunoreactive neurons and minimal activity for acetylcholinesterase and tyrosine hydroxylase), unlike the adjacent putamen and ventral pallidum. A putative endopiriform nucleus can be identified in the interior of the piriform lobe of the echidna as calretinin immunoreactive neurons embedded within the white matter. The situation is much less clear in the platypus, but our data suggest that there may be an insular claustrum deep to frontal cortex, separated from layer VI by only a thin layer of white matter. We could not identify an endopiriform nucleus in our platypus material. Our findings indicate that presence of the claustrum cannot be considered a feature confined to therian mammals and lend weight to arguments that this structure was present in the ancestral mammalian brain.