Fig. 3
Subtracted brain regions expressing putative valine amide peptide immunoreactivity. P18C5-mAb-ir detected along the neuroaxis of the rat brain (Fig. 7a–e) was subtracted from brain areas expressing peptide immunoreactivity for α-MSH, metorphamide, secretin, and urocortins 1-2. As shown, moderate-to-high intensity mAb-ir signals detected in subtracted brain areas were localized to the cortex [secondary motor (M2), the hindlimb (S1HL), the trunk region (S1Tr) of the primary somatosensory, parietal association (PtA), secondary visual mediomedial (V2MM) and mediolateral (V2ML) cortices] and the main and accessory olfactory nuclei [anterior accessory olfactory nucleus dorsal (AOD), nucleus ventral (AOV), nucleus lateral (AOL) and nucleus medialis (AOM), the olfactory tubercle (Tu), the bed nucleus of the accessory olfactory tract (BAOT) and the nucleus of the lateral olfactory tract (LOT)]. Subcortical areas displaying mAb-ir signal included the subiculum (S), the core (AcbC) and shell (AcbSh) of the nucleus accumbens (NACC), the substantia innominata of the septum (SI), the amygdalo-hippocampal (Ahi) and amygdalo-anterior nuclei (AA) of the limbic system, the lateral posterior nucleus of the thalamus (LPMR) and the lateral hypothalamic (LH) nuclei. Brainstem areas included the substantia nigra (SNL), the superior colliculus (SC), the retrorubral nucleus (RR), the solitary tract (Sol) and the flocculus of the cerebellum (FL). With the exception of the areas described above, only the LH and the FL showed intense immunolabeling similar to that detected in cortical layers of the Cb, the IL of the pituitary and the adrenal medulla (me). As shown, these brain areas were arbitrarily graded as displaying low Val-CONH2-peptide immunoreactive signals because they corresponded to areas containing the amidated peptides used as references in this study (see text for additional details). The neuroanatomical areas showing putative valine amide peptide immunoreactivity were identified based on the rat brain atlas of Paxinos and Watson [46]