Figure 5. Colocalization of MCH neuropeptides and GAD₆₇ mRNA in neurons of the tuberal hypothalamus.

(A) Photomicrograph showing one MCH-ir cell (black star, brown cytoplasmic staining only) and one MCH-ir cell co-expressing GAD₆₇ mRNA (black arrows, brown and blue diffuse cytoplasmic stainings superimposed) in the lateral hypothalamic area. (B) Graphics illustrating the distribution of MCH-ir cells along the rostrocaudal axis (from the first to the 7^th section). The upper graph reports the average number of MCH-ir cells (black dots) (median ± Q1;Q3) and of MCH-ir/GAD⁺ neurons (red dots) counted on hemisections from three rats. The bottom graph illustrates the percent of MCH-ir/GAD⁺ cells over the total number of MCH-ir neurons on each hemisection. (C) Schematic distribution of MCH-ir (black dots) and MCH-ir/GAD⁺ (red dots) neurons in on representative rat brain. The approximate planes of sections relative to Bregma (β), according to the atlas of Paxinos and Watson [51], are −2.10 (1), −2.40 (2), −2.70 (3), −3.00 (4), 3.30 (5), 3.60 (6) and −3.90 (7). Scale bar: 30 µm. Abbreviations: AH: anterior hypothalamic area; Arc: arcuate nucleus; DA: dorsal hypothalamic area; DM: dorsomedial hypothalamic nucleus; f: fornix; ic: internal capsule; LH: lateral hypothalamic area; ml: medial lemniscus; mt: mammillothalamic tract; PH: posterior hypothalamic area; PMV: premammillary nucleus, ventral part; PVN: paraventricular hypothalamic nucleus; opt: optic tract; Rt: reticular thalamic nucleus; SPF: subparafascicular thalamic nucleus; STh: subthalamic nucleus; SubI: subincertal nucleus; VM: ventromedial thalamic nucleus; VMH: ventromedial hypothalamic nucleus; ZI: zona incerta.