Stimulation of the Periaqueductal Gray matter (PAG) and nucleus retroambiguus (nRA) evokes vocalization. nRA modulates the activity of laryngeal motoneurons located in the nucleus ambiguus (nA). We have demonstrated the involvement of rostral and ventral pontine structures in altering laryngeal caliber. A heightened expression of the FOXP2 protein was described in both PAG and pontine regions implicated in cardiorespiratory control.To investigate the interactions between the dlPAG and neurons of the nA assessing the double staining patterns of c-Fos/FOXP2 protein immunoreactivity (c-Fos-ir/FOXP2-ir) across the rostro-caudal extent of the nA.Experimental studies were carried out with non-inbred male rats (n=10), SPF, Sprague-Dawley (250-300 g) housed under standard conditions and anesthetized with sodium pentobarbitone (60 mg/kg i.p., initial dose, supplemented 2mg/ kg, i.v., as necessary). The pattern of c-Fos-ir/FOXP2-ir and double immunofluorescence staining were examinated throughout the nA region during electrical stimulation of the dlPAG (1 ms pulses, ranging from 30-50 μA, at 100 Hz, during 5 s). Guanethidine (10 mg/kg iv) was administered to suppress sympathetically mediated cardiovascular responses. Only data from animals in which the histology showed that the microelectrodes were positioned within the dlPAG were used for statistical procedures.dlPAG stimulation induced an increase in c-Fos-ir, with ipsilateral predominance in both the loose (p<0.05) and compact formation (p<0.01) within the nA. Furthermore, we could confirm the bilateral expression of FoxP2 across all nA domains.Our study contributes with new data on the role of the mesencephalic neuronal circuits in the control mechanisms of subglottic pressure and laryngeal activity.