Orexin A activates retrotrapezoid neurons in mice

The retrotrapezoid nucleus (RTN), located at the ventral surface of the medulla oblongata, contains glutamatergic Phox2b-expressing interneurons that have central respiratory chemoreceptor properties. RTN also operates as a relay for hypothalamic pathways that regulate breathing, one of which probably originates from the orexinergic neurons (Dias et al., 2009. J. Physiol. 587, 2059–2067). The present study explores this hypothesis at the cellular level. Using immunohistochemistry in adult Phox2b-eGFP transgenic mice, we demonstrate the presence of numerous close appositions between orexin-containing axonal varicosities and RTN chemoreceptor neurons. Using electrophysiological recordings in slices from neonatal (P6-P10) Phox2b-eGFP mice, we show that orexin A produces a robust dose-dependent excitation of the acid-sensitive RTN neurons (ED50 ∼250 nM). These data support the idea that RTN neurons are a point of convergence for several groups of CNS neurons that contribute to respiratory chemoreflexes, now including serotonergic and orexinergic neurons.