Immunohistochemical study on the neuronal diversity and three-dimensional organization of the mouse entopeduncular nucleus

• Entopeduncular nucleus of mouse has parvalbumin-rich core and substance P-rich shell.
• Parvalbumin neurons known to target thalamus constitute only 30% of EPN neurons.
• New type of GABAergic neurons lacking both parvalbumin and somatostatin are found.
• About 10% of neurons in mouse entopeduncular nucleus is non-GABAergic.

The entopeduncular nucleus (EPN) is one of the major output nuclei of the basal ganglia in rodents. Previous studies have divided it into rostral and caudal halves, with the former containing somatostatin (SOM)-immunoreactive neurons and the latter dominated by parvalbumin (PV)-containing neurons, respectively. However, it is unclear whether this simple rostrocaudal segmentation is appropriate, and the possibility of the existence of other neuronal populations remains to be investigated. In this study the cytoarchitecture of the mouse EPN was analyzed immunohistochemically. Substance P (SP)-immunoreactivity determined the extent of the EPN, which was 800 μm-long along the rostrocaudal axis. PV-positive neurons were concentrated in the caudal two-thirds of this range. PV-negative neurons were abundant in the rostral half but were further located caudally around the PV neuron-rich core. PV(+)/SOM(−) and PV(−)/SOM(+) neurons constituted 28.6% and 45.7% of EPN neurons, respectively, whereas the remaining population (25.7%) exhibited neither immunoreactivity. Eleven percent of EPN neurons lacked immunoreactivity for glutamic acid decarboxylase, indicating their non-GABAergic nature. Three-dimensional reconstruction revealed that PV-rich/SP-poor core was surrounded by PV-poor/SP-rich shell region. Therefore, presumptive thalamus-targeting PV neurons are outnumbered by other populations, and the regional heterogeneity shown here might be related to functionally distinct pathways through the basal ganglia.