Rostral ventral medulla (RVM) contains significant numbers of local GABAergic neurons which may subserve respiratory chemosensory and baroreceptor reflexes. Nicotinic mechanisms stimulate release of GABA in certain brainstem neurons. Whether the GABAergic neurons at RVM express nicotinic cholinergic receptors (nAChRs) is not known. We used glutamic acid decarboxylase 67-kDa isoform (GAD67) and parvalbumin (PV) as anatomical markers to identify the GABAergic neurons of the RVM and caudal pons and performed double labeling to evaluate the expression of alpha-7 and alpha-4 nAChRs by GAD67 and PV-imnunoreactive (ir) cells at these sites. GAD67-ir cells were found at the ventrolateral pontomedullary border in areas adjacent to the A5 noradrenergic cell group and overlapping the facial nucleus lateral subnuclei and para-facial zones. Of 205 GAD67-ir cells labeled at these sites, 74% exhibited immunoreactivity for alpha-7 nAChRs. Alpha-4 immunoreactivity was also present in 35% of GAD67-ir cells at these sites. The PV-ir cells of RVM and caudal pons were found medial to the facial nucleus and lateral to the pyramid in a column distinct from the GAD67-ir cells. Virtually all the PV-ir cells demonstrated immunoreactivity for alpha-4 nAChR (95%) and alpha-7 (93%) subunits of nAChRs. Differential expression of GAD67 and PV by neurons at the pontomedullary border implies that PV may not be a valid marker for GABAergic neurons. The expression of alpha-4 and alpha-7 nAChRs by GAD67-ir cells suggests nicotinic cholinergic modulation of GABAergic signaling at these ventrolateral pontomedullary sites.