Quantitative and immunohistochemical analysis of neuronal types in the mouse caudal nucleus tractus solitarius: Focus on GABAergic neurons

γ-Aminobutyric acid-ergic (GABAergic) neurons are major inhibitory interneurons that are widely distributed in the central nervous system. The caudal nucleus tractus solitarius (cNTS), which plays a key role in respiratory, cardiovascular, and gastrointestinal function, contains GABAergic neurons for regulation of neuronal firing. In the present study, GABAergic neuronal organization was analyzed in relation to the location of subnuclei in the mouse cNTS. According to the differential expression of glutamate decarboxylase 67 (GAD67), vesicular glutamate transporter 2 (VGLUT2), calbindin, and tyrosine hydroxylase (TH) mRNAs, the cNTS was divided into four subnuclei: the subpostrema, dorsomedial, commissural, and medial subnuclei. The numerical density and size of soma in the four subnuclei were then quantified by an unbiased disector analysis. Calbindin-positive cells constituted subpopulations of small non-GABAergic neurons preferentially localized in the subpostrema subnucleus. TH-positive cells constituted large neurons preferentially localized in the medial subnucleus. GABAergic neurons constituted a subpopulation of small neurons, preferentially localized in the commissural and medial subnuclei, which represented ≥50% of small cells in these subnuclei. Thus, the GABAergic small neurons were located around TH-positive large cells in the ventrolateral portion of the cNTS. This finding, in combination with results of previous studies in the rat cNTS showing that large cells originate efferents from the cNTS, suggests that GABAergic small neurons in the commissural and medial subnuclei might regulate output from the cNTS.