Orexin neurons in the hypothalamus mediate cardiorespiratory responses induced by disinhibition of the amygdala and bed nucleus of the stria terminalis.

We previously showed that the defense response elicited by stressors was attenuated in prepro-orexin knockout mice and in orexin neuron-ablated mice, and we proposed that orexin serves as a master switch within multiple efferent pathways that mediate the defense response. In this study we sought to determine whether excitation of the amygdala (AMG) or the bed nucleus of stria terminalis (BNST) activates orexin-containing neurons and whether those neurons are essential in eliciting cardiorespiratory responses to the stimulus. In urethane-anesthetized mice, the GABA-A receptor antagonist bicuculline was microinjected into the AMG or BNST and blood pressure, heart rate, and respiration were measured. Injection of bicuculline in either site induced long-lasting dose-dependent cardiorespiratory excitation in wild-type mice. In contrast, mice in which orexin neurons had been ablated demonstrated no such response after activation of the AMG and an attenuated response after activation of the BNST. Double immunohistochemical staining for orexin and c-Fos, an indicator of neural activation, revealed that an injection of bicuculline induced significantly larger numbers of orexin positive neurons that expressed c-Fos in the perifornical/dorsomedial hypothalamus (58.2+/-6.4% into AMG and 66.4+/-6.6% into BNST, n=3 each) than did vehicle (18.2+/-4.4% into AMG and 28.3+/-2.1% into BNST). Disinhibition to the BNST induced widespread expression of c-Fos not only in orexin-containing neurons but also other neurons in the hypothalamus. We conclude that orexin-containing neurons in the medial hypothalamus mediate at least a part of AMG- and BNST-induced cardiorespiratory responses.