Orexin facilitates the peripheral chemoreflex via corticotropin releasing hormone neurons projecting to the nucleus of the solitary tract.

We previously showed that orexin neurons are activated by hypoxia and facilitate the peripheral chemoreflex (PCR)-mediated hypoxic ventilatory response (HVR), mostly by promoting the respiratory frequency response. Orexin neurons project to the nucleus of the solitary tract (nTS) and the paraventricular nucleus of the hypothalamus (PVN). The PVN contributes significantly to the PCR and contains nTS-projecting corticotropin releasing hormone (CRH) neurons. We hypothesized that in male rats orexin neurons contribute to the PCR by activating nTS-projecting CRH neurons. We used neuronal tract tracing and immunohistochemistry (IHC) to quantify the degree that hypoxia activates PVN-projecting orexin neurons. We coupled this with orexin receptor (OxR) blockade with suvorexant (Suvo, 20 mg/kg, i.p.) to assess the degree that orexin facilitates the hypoxia-induced activation of CRH neurons in the PVN, including those projecting to the nTS. In separate groups of rats, we measured the PCR following systemic orexin 1 receptor (Ox1R) blockade (SB-334867;1 mg/kg) and specific Ox1R knockdown in PVN. OxR blockade with Suvo reduced the number of nTS and PVN neurons activated by hypoxia, including those CRH neurons projecting to nTS. Hypoxia increased the number of activated PVN-projecting orexin neurons but had no effect on the number of activated nTS-projecting orexin neurons. Global Ox1R blockade and partial Ox1R knockdown in the PVN significantly reduced the PCR. Ox1R knockdown also reduced the number of activated PVN neurons and the number of activated tyrosine-hydroxylase neurons in the nTS. Our findings suggest orexin facilitates the PCR via nTS-projecting CRH neurons expressing Ox1R.Significance Statement Previously we showed that orexin contributes to the peripheral chemoreflex (PCR), but the mechanisms underlying this effect remain unknown. Here we show that: 1) orexin receptor blockade reduces the activation of the PVN and nTS; 2) hypoxia activates orexin neurons that project to the PVN, but not those projecting to nTS; 3) orexin receptor blockade reduces the activation of nTS-projecting corticotrophin releasing hormone (CRH) neurons in the PVN; 4) orexin 1 receptor (Ox1R) blockade and specific Ox1R knockdown in the PVN reduce the strength of the PCR, and 5) Ox1R knockdown reduces the number of activated PVN neurons and tyrosine-hydroxylase neurons in the nTS. These findings suggest that PVN-projecting orexin neurons facilitate the PCR via Ox1R on nTS-projecting CRH neurons.