The nucleus of the tractus solitarius (NTS) is a primary termination zone for laryngeal, gustatory, cardiovascular, respiratory, gastrointestinal, and other visceral afferents. Although considerable information is available on the neurochemical aspects of the NTS in general, very little is known about glutamate receptors that may underlie many of the different functions mediated by the NTS. In addition, most previous glutamate receptor distribution studies were performed in the rat, whereas the cat, the subject of many physiological experiments involving the NTS, has received little attention. In the present study, the immunohistochemical distribution of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)-selective glutamate receptor subunits (GluR1, GluR2/3, GluR4) and the N-methyl-D-aspartate (NMDA) receptor subunit NR1 in the cat caudal brainstem was investigated by using subunit-specific antibodies. In the NTS, statistically significant differences were seen in the distribution of each antibody. Highest labeling was seen for GluR2/3 in most subnuclei, whereas GluR1-immunoreactive neurons were found more frequently than were NR1- or GluR4-immunoreactive neurons. GluR1 immunolabeling was particularly high in the interstitial subnucleus, whereas GluR2/3 immunolabeling was particularly high in the intermediate subnucleus. Qualitatively, labeling for GluR4 was most common in glia. The present results indicate that glutamate receptors show different subunit distributions in the subnuclei of the NTS and in other adjacent structures. This finding suggests that neurons in these structures are designed to respond differently to excitatory input.

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The main functions of the larynx are protection of the airways, respiration, and vocalization. Previous studies have suggested a link between the mechanisms controlling vocalization and afferent feedback from the larynx. We inquired whether stimulation of the laryngeal afferents that run in the internal branch of the superior laryngeal nerve (ISLN) activates neurons of the periaqueductal gray (PAG), a midbrain region implicated in vocalization. We counted the number of neurons expressing Fos, the protein product of the immediate early gene c-fos, in the PAG. The counts were done both in experimental cats after electrical stimulation of the ISLN and nonstimulated controls. We also investigated the possible presence of nitric oxide synthase, an enzyme that synthesizes nitric oxide, in PAG neurons that respond to laryngeal afferent stimulation by double labeling for reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase and Fos. Fos expression was significantly greater (P < or = 0.00714) in the lateral and dorsolateral regions of the PAG in the experimental group than in the controls. The Fos-immunoreactive neurons did not contain NADPH-diaphorase, a marker for nitric oxide synthase. Our study suggests that laryngeal afferent stimulation activates neurons in discrete longitudinal columns of the PAG including the regions that have previously been shown to be involved in vocalization, and that these neurons do not contain nitric oxide synthase.