Do Trigeminal Nerves that Innervate the Nose and External Nares Project to Medullary Dorsal Horn Locations Activated during Voluntary Diving in Rats?

Abstract

Underwater diving initiates the diving response, an autonomic cardiorespiratory reflex that includes parasympathetically‐mediated bradycardia, a sympathetically‐mediated increase in total peripheral resistance, and apnea. The initial afferent node of the reflex circuitry includes the ventral tip of the medullary dorsal horn (MDH), between the pyramidal decussation and obex. This region is also the location of the central terminations of trigeminal nerves that innervate the nose and external nares. Accordingly, the objective of this research was to compare the pattern of neuronal activation of secondary neurons activated by voluntary diving, as determined by neuronal Fos production, with the central terminations of specific trigeminal nerves that innervate the nose and external nares, as determined by the transganglionic tracer Wheat Germ Agglutinin (WGA). Sprague‐Dawley rats (N=12) were trained to voluntarily dive through a 5 m underwater maze. Six rats then received bilateral AEN sectioning, while 6 rats received sham surgery. Nine days post‐surgery all rats had 24 dive trials over 2 hours to activate brainstem neurons to produce Fos. Other rats had 1 μl of 100% WGA injected into the anterior ethmoidal nerve (AEN; N=4), the main orbital branch of the infraorbital nerve (ION; N=7), terminal branches of the ION that innervate the nasal region (N=9), or supratrochlear nerve (N=8). Results indicate Fos was primarily located within the ventral tip of the superficial MDH and adjacent paratrigeminal nucleus, between the pyramidal decussation caudally and the obex rostrally. The Fos labeling was heaviest when the area postrema was present, especially when Spinal Trigeminal Nucleus Interpolaris displaces the ventral MDH medially. This pattern of Fos labeling within the MDH was similar in rats both with and without intact AENs. Within the MDH WGA central termination sites showed varying degrees of overlap with the location of Fos‐positive neurons. The AEN WGA labeling had the best overlap with Fos‐positive neurons. The main orbital branch of the ION had extensive WGA labeling throughout the MDH, both ventrally and dorsally, and did partially overlap the MDH Fos labeling. The WGA labeling from both nasal branches of the ION and supratrochlear nerve were located more caudally within the MDH near the pyramidal decussation, thus showing poor overlap with the Fos labeling. We conclude that the AEN, and possibly the ION, provides the anatomical pathway that provides the afferent information necessary to initiate the mammalian diving response. However, in the absence of the AEN, it is not clear whether the ION can actually provide this afferent information.Support or Funding InformationSupported by MWU Intramural funding (PFM), CCOM Suarez Summer Research Fellowships (BDP and KAL), and CHS MBS program (EAW).