Ann M. Thompson

Serotonin and Serotonin Receptors in the Central Auditory System

Immunohistochemical and ligand-binding techniques were used to visualize the neurotransmitter serotonin and one of its receptors, the 5-HT1A subtype, in auditory nuclei of the brainstem. Serotonergic fibers and terminal endings were found in all auditory nuclei extending from the cochlear nucleus to the inferior colliculus, including the superior olivary complex and the nuclei of the lateral lemniscus. The density of the innervation varied between and within each nucleus. All serotonergic cell bodies were located outside the auditory nuclei. The 5-HT1A receptor subtype was found in the cochlear nucleus as well as in the inferior colliculus. With no serotonergic cell bodies present in the auditory nuclei, the present neuroanatomic and neurochemical findings support behavioral and neurophysiologic findings that the serotonergic system may modulate central auditory processing.

Serotonin in the inferior colliculus

It has been recognized for some time that serotonin fibers originating in raphe nuclei are present in the inferior colliculi of all mammalian species studied. More recently, serotonin has been found to modulate the responses of single inferior colliculus neurons to many types of auditory stimuli, ranging from simple tone bursts to complex species-specific vocalizations. The effects of serotonin are often quite strong, and for some neurons are also highly specific. A dramatic illustration of this is that serotonin can change the selectivity of some neurons for sounds, including species-specific vocalizations. These results are discussed in light of several theories on the function of serotonin in the IC, and of outstanding issues that remain to be addressed.

Serotonergic innervation of the auditory brainstem of the Mexican free-tailed bat, Tadarida brasiliensis

Anatomical and electrophysiological evidence suggests that serotonin alters the processing of sound in the auditory brainstem of many mammalian species. The Mexican free‐tailed bat is a hearing specialist, like other microchiropteran bats. At the same time, many aspects of its auditory brainstem are similar to those in other mammals. This dichotomy raises an interesting question regarding the serotonergic innervation of the bat auditory brainstem: Is the serotonergic input to the auditory brainstem similar in bats and other mammals, or are there specializations in the serotonergic innervation of bats that may be related to their exceptional hearing capabilities? To address this question, we immunocytochemically labeled serotonergic fibers in the brainstem of the Mexican free‐tailed bat, Tadarida brasiliensis. We found many similarities in the pattern of serotonergic innervation of the auditory brainstem in Tadarida compared with other mammals, but we also found two striking differences. Similarities to staining patterns in other mammals included a higher density of serotonergic fibers in the dorsal cochlear nucleus and in granule cell regions than in the ventral cochlear nucleus, a high density of fibers in some periolivary nuclei of the superior olive, and a higher density of fibers in peripheral regions of the inferior colliculus compared with its core. The two novel features of serotonergic innervation in Tadarida were a high density of fibers in the fusiform layer of the dorsal cochlear nucleus relative to surrounding layers and a relatively high density of serotonergic fibers in the low‐frequency regions of the lateral and medial superior olive. J. Comp. Neurol. 435:78–88, 2001.

Serotonin projection patterns to the cochlear nucleus.

The cochlear nucleus is well known as an obligatory relay center for primary auditory nerve fibers. Perhaps not so well known is the neural input to the cochlear nucleus from cells containing serotonin that reside near the midline in the midbrain raphe region. Although the specific locations of the main, if not sole, sources of serotonin within the dorsal cochlear nucleus subdivision are known to be the dorsal and median raphe nuclei, sources of serotonin located within other cochlear nucleus subdivisions are not currently known. Anterograde tract tracing was used to label fibers originating from the dorsal and median raphe nuclei while fluorescence immunohistochemistry was used to simultaneously label specific serotonin fibers in cat. Biotinylated dextran amine was injected into the dorsal and median raphe nuclei and was visualized with Texas Red, while serotonin was visualized with fluorescein. Thus, double-labeled fibers were unequivocally identified as serotoninergic and originating from one of the labeled neurons within the dorsal and median raphe nuclei. Double-labeled fiber segments, typically of fine caliber with oval varicosities, were observed in many areas of the cochlear nucleus. They were found in the molecular layer of the dorsal cochlear nucleus, in the small cell cap region, and in the granule cell and external regions of the cochlear nuclei, bilaterally, of all cats. However, the density of these double-labeled fiber segments varied considerably depending upon the exact region in which they were found. Fiber segments were most dense in the dorsal cochlear nucleus (especially in the molecular layer) and the large spherical cell area of the anteroventral cochlear nucleus; they were moderately dense in the small cell cap region; and fiber segments were least dense in the octopus and multipolar cell regions of the posteroventral cochlear nucleus. Because of the presence of labeled fiber segments in subdivisions of the cochlear nucleus other than the dorsal cochlear nucleus, we concluded that the serotoninergic projection pattern to the cochlear nucleus is divergent and non-specific. Double-labeled fiber segments were also present, but sparse, in the superior olive, localized mainly in periolivary regions; this indicated that the divergence of dorsal and median raphe neurons that extends throughout regions of the cochlear nucleus also extended well beyond the cochlear nucleus to include at least the superior olivary complex as well.

Heterogeneous projections of the cat posteroventral cochlear nucleus

The anterograde tracer Phaseolus vulgaris‐leucoagglutinin was used to identify the projections of the posteroventral cochlear nucleus in cats. After labeling predominately cells of the core and multipolar regions, varicose fibers were observed in a variety of auditory nuclei. Ipsilaterally, most varicose fibers were located in periolivary regions situated lateral to the medial superior olive of the superior olivary complex. Contralaterally, the majority of labeled fibers were located in the ventral nucleus of the trapezoid body and the ventral nucleus of the lateral lemniscus. Labeled varicose fibers were also observed in regions not commonly identified as receiving input from the posteroventral cochlear nucleus. These regions included bilaterally the principal nuclei of the superior olivary complex, some periolivary regions, and the sagulum, as well as the ipsilateral intermediate and dorsal nucleus of the lateral lemniscus, inferior colliculus, and lateral pontine nucleus. Both similarities and differences were observed in the projections of the core and multipolar regions. With the exception of calyceal‐type endings in the contralateral ventral nucleus of the lateral lemniscus, the varicose fibers in all regions, including the contralateral medial nucleus of the trapezoid body, were beaded, en passant type terminal varicosities. J. Comp. Neurol. 390:439–453, 1998.

Light microscopic evidence of serotoninergic projections to olivocochlear neurons in the bush baby (Otolemur garnettii)

Double-label techniques were used to concomitantly label olivocochlear neurons and serotoninergic fibers in the bush baby (Otolemur garnettii) brainstem. Light-microscopic examination (using a 100 x plan apochromatic oil-immersion objective) of the sections revealed that serotonin-positive varicosities (presumptive terminal endings) contacted somata and dendrites of neurons belonging to both the lateral and medial olivocochlear neurons near the superior olivary nuclei. These results provide direct evidence that the olivocochlear system (a specific auditory brainstem pathway) receives input from the serotoninergic system (a diffuse reticular brainstem network).

Serotoninergic innervation of stapedial and tensor tympani motoneurons

Retrograde tracing and neurotransmitter immunohistochemistry were combined to determine whether serotonin neurons innervated stapedial and tensor tympani motoneurons. With high-power light microscopy, putative axo-somatic and axo-dendritic contacts were observed between serotonin-positive endings and both stapedial and tensor tympani motoneurons, indicating that serotonin neurons terminate on brainstem motoneurons innervating the middle-ear muscles. With this connection, the serotonin system may directly modulate middle-ear muscle activity.

Neural connections identified with PHA-L anterograde and HRP retrograde tract-tracing techniques.

This paper describes a method of identifying specific input and output elements of a two-neuron projection pathway. Phaseolus vulgaris leucoagglutinin (PHA-L) anterograde tract-tracing was used in combination with the retrograde transport of horseradish peroxidase (HRP) to demonstrate connections between small groups of neurons in the brainstem auditory system. Specifically, the projection from cochlear nucleus to olivary neurons that project to the cochlea were demonstrated. The first neuron in this pathway (the cochlear nucleus neuron) was anterogradely labelled with PHA-L and could be traced via labelled axons and terminals to the second neuron (the olivocochlear neuron) whose soma was labelled with HRP after cochlear injection.

Activation of serotonergic neurons during salicylate-induced tinnitus.

Hypothesis: Serotonergic neurons are activated during salicylate-induced tinnitus and modulate the cochlea during tinnitus. Background: During salicylate-induced tinnitus in the gerbil, neurons in the dorsal raphe nucleus were activated. Because approximately half of the neurons in this region are serotonergic, this indicates that serotonin (5-HT) might play a role in the mechanisms of central tinnitus. The goal of this study was to determine if serotonergic neurons are activated during salicylate-induced tinnitus. Furthermore, to determine if the same neurons might modulate the cochlea during tinnitus, neuroanatomic tract-tracing with 5-HT immunohistochemistry was used to determine if serotonergic neurons project to the gerbil cochlea. Methods: A randomized, prospective study was performed. Six gerbils were injected with salicylate (saline for controls). Four hours later, the gerbils were euthanized and perfused, and their brains were collected for immunohistochemical labeling of 5-HT and c-fos. For the tract-tracing, FluoroGold was injected into the cochleae of 3 gerbils. The gerbils were euthanized and perfused 4 to 11 days later, and the brains immunohistochemically were processed for 5-HT. Results: More serotonergic neurons expressed c-fos in the salicylate-injected animals compared with the controls. The increase was significant for 3 of the 8 major serotonergic cell groups including B7, B9, and the caudal linear nucleus. Despite robust labeling of olivocochlear and vestibular efferents with FluoroGold, 5-HT-labeled neurons containing FluoroGold were lacking. Conclusion: Salicylate-induced tinnitus activates serotonergic neurons in rostral cell groups. Activation of these neurons is not likely to influence cochlear function directly but is likely to influence a number of auditory and nonauditory regions known to be involved with tinnitus.

Aminergic projections to cochlear nucleus via descending auditory pathways

The cochlear nucleus (CN) receives descending input from a variety of auditory nuclei. Descending inputs from the superior olive in particular have been well described, especially those of olivocochlear neurons, which terminate ultimately in the cochlea. It has been demonstrated that olivocochlear neurons receive serotonergic and noradrenergic inputs and thus form a route by which the aminergic system may modulate cochlear mechanisms. Since olivocochlear neurons send collaterals into the CN, it is possible that they also from a route by which the aminergic systems modulate CN processes. The goal of the current study was to determine if neurons in the superior olive that projected to the CN received serotonergic or noradrenergic inputs. The retrograde tracer WGAapoHRP-Au was injected into the CN of cats. The brainstems were silver-enhanced to visualize the tracer and then immunohistochemically processed with antibodies raised against serotonin or dopamine-beta-hydroxylase (DBH) to label serotonergic or noradrenergic fibers, respectively. The sections were viewed with high power light microscopy to determine if the retrogradely labeled neurons were contacted by serotonin- or DBH-immunoreactive varicosities. Retrogradely labeled cells were observed in auditory brainstem nuclei known to project to the CN including the superior olivary complex and inferior colliculus bilaterally and the opposite CN. In these regions, retrogradely labeled neurons were closely associated with serotonin- and/or DBH-immunoreactive varicosities. Assuming a synaptic relationship between the projection neurons and varicosities, these results indicate that the serotonergic and noradrenergic systems innervate the descending pathways to the CN. Since the serotonergic and noradrenergic systems modulate their targets based on level of arousal, these results support the theory that descending systems are involved in selective attention.