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Somatostatin and leu-enkephalin in the rat auditory brainstem during fetal and postnatal development.

A transient expression of the neuropeptide somatostatin has been described in several brain areas during early ontogeny and several opioid peptides, such as leu-enkephalin, have also been found in the brain at this stage in development. It is therefore believed that somatostatin and leu-enkephalin may play a role in neural maturation. The aim of the present study was to describe the spatiotemporal pattern of somatostatin and leu-enkephalin immunoreactivity in the auditory brainstem nuclei of the developing rat and to correlate it with other developmental events. In order to achieve this goal, we applied peroxidase-antiperoxidase immunocytochemistry to rat brains between embryonic day (E) 17 and adulthood. Somatostatin immunoreactivity (SIR) was found in all nuclei of the auditory brainstem, yet it was temporally restricted in most nuclei. SIR appeared prenatally and reached maximum levels around postnatal day (P) 7, when great numbers of immunoreactive neurons were present in the ventral cochlear nucleus (VCN) and in the lateral lemniscus. At that time relatively low numbers of cells were labeled in the dorsal cochlear nucleus, the lateral superior olive (LSO), and the inferior colliculus (IC). During the same period, when somata in the VCN were somatostatin-immunoreactive (SIR), a dense network of labeled fibers was also present in the LSO, the medial superior olive (MSO), and the medial nucleus of the trapezoid body (MNTB). As these nuclei receive direct input from VCN neurons, and as the distribution and morphology of the somatostatinergic fibers in the superior olivary complex (SOC) was like that of axons from VCN neurons, these findings suggest a transient somatostatinergic connection within the auditory system. Aside from the LSO, MSO, and MNTB, labeled fibers were found to a smaller extent in all other auditory brainstem nuclei. After P7, the SIR decreased and only a few immunoreactive elements were found in the adult auditory brainstem nuclei, indicating that somatostatin is transiently expressed in the rat auditory brainstem. Leu-enkephalin immunoreactivity showed a lower number and weaker intensity of labeled structures as compared to SIR, with E18 being the earliest day at which labeled fibers appeared in the SOC. At birth, immunoreactive fibers were also present in the cochlear nuclear complex and in the IC. Leu-enkephalin immunoreactive somata were found only after P12 in the CN and after P16 in the IC. Leu-enkephalin immunoreactivity was not transient, but increased progressively with age until about P21, when the adult levels were reached.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Neurochemistry of the peripheral and central auditory system after ototoxic drug exposure: implications for tinnitus.

Platinum-containing drugs, such as cisplatin and carboplatin, are known to have ototoxic side effects causing hearing loss that may be accompanied by tinnitus. This study reviews recent studies on the ototoxicity of cisplatin and carboplatin and summarizes the effects of protective agents that may prevent hearing loss and tinnitus. The primary locus of ototoxicity is in the cochlea, but oxidative stress to the inferior colliculus has been reported recently with carboplatin. Enhanced spontaneous activity within the dorsal cochlear nucleus has been correlated with loss of outer hair cells in animal experiments using cisplatin. This may result from disinhibition of neurons within the dorsal cochlear nucleus caused by reduced input from spiral ganglion cells. Carboplatin may cause tinnitus by oxidative stress within the inferior colliculus or by loss of inhibition within the inferior colliculus resulting from cochlear damage. This could lead to compensatory gain and enhanced responses in neurons within the auditory cortex. Protective agents may prevent tinnitus by preventing damage to the cochlea, thereby obviating the development of disinhibition within central auditory pathways.

Animals↗

Distribution of primary cochlear afferents in the bulbar nuclei of the rat: a horseradish peroxidase (HRP) study in parasagittal sections.

HRP was injected into the cochleae of 25 young albino rats in order to trace the primary afferents to the bulbar cochlear nuclei. Besides the classic V-shaped pattern and unconnected with it, HRP labelling revealed two plexuses stemming directly from the axons of the cochlear root. The plexuses cover the posterior area of the posteroventral cochlear nucleus (posterior plexus) and the anterolaterodorsal area of the anteroventral cochlear nucleus (anterior plexus). The fibres giving rise to these two plexuses were previously grouped in two bundles which have been called the posterior and anterior bundles, respectively. The origin of the anterior bundle is typically seen with the fibres stemming out at right angles; the origin and course of the posterior bundle, which characteristically cross over, is also a typical feature.

Afferent Pathways↗

Multiple projections from the ventral nucleus of the trapezoid body in the rat.

An analysis of the central projections of the ventral nucleus of the trapezoid body (VNTB) in the rat, a region of the superior olivary complex known for its neuronal heterogeneity, was made using two anterograde axonal tracers, [3H]leucine and biotinylated dextran amine (BDA). A mixture of these tracers was injected iontophoretically into the VNTB and the results analyzed by first assessing magnitudes of autoradiographic signal in nuclei receiving projections and then identifying the axons and terminals responsible for this signal in parallel sets of sections processed for BDA. Our analysis showed that in addition to its projections to each cochlea via the olivocochlear bundle, the VNTB has 3 major central sites of axonal terminations: (1) the cochlear nucleus, particularly the molecular layer of the contralateral dorsal cochlear nucleus, (2) the contralateral lateral superior olive, and (3) the ipsilateral inferior colliculus. Other sites receiving projections from the VNTB included the VNTB itself and the nuclei of the lateral lemniscus. Significantly, the relative magnitudes of labeling within the nuclei receiving inputs from the VNTB varied consistently as a function of the dorsoventral location of the injection site, confirming previous work showing that there is a partial segregation within this nucleus of neurons according to their projections. Our data also revealed an orderly topographic pattern of projections to the cochlear nuclei, lateral superior olive and the inferior colliculus which is consistent with the known tonotopic organization both of the VNTB and these projection targets. Methodologically, the co-injection of two tracers was advantageous in that patterns of silver grains in autoradiographs could be used to confirm whether axons and terminals labeled with BDA had originated from labeled somata at the injection site or were the result of uptake of BDA by fibers of passage.

Animals↗

A computer model of amplitude-modulation sensitivity of single units in the inferior colliculus.

A computer model is presented of a neural circuit that replicates amplitude-modulation (AM) sensitivity of cells in the central nucleus of the inferior colliculus (ICC). The ICC cell is modeled as a point neuron whose input consists of spike trains from a number of simulated ventral cochlear nucleus (VCN) chopper cells. Input to the VCN chopper cells is provided by simulated spike trains from a model of the auditory periphery [Hewitt et al., J. Acoust. Soc. Am. 91, 2096-2109 (1992)]. The performance of the model at the output of the auditory nerve, the cochlear nucleus and ICC simulations in response to amplitude-modulated stimuli is described. The results are presented in terms of both temporal and rate modulation transfer functions (MTFs) and compared with data from physiological studies in the literature. Qualitative matches were obtained to the following main empirical findings: (a) Auditory nerve temporal-MTFs are low pass, (b) VCN chopper temporal-MTFs are low pass at low signal levels and bandpass at moderate and high signal levels, (c) ICC unit temporal-MTFs are low pass at low signal levels and broadly tuned bandpass at moderate and high signal levels, and (d) ICC unit rate-MTFs are sharply tuned bandpass at low and moderate signal levels and flat at high levels. VCN and ICC units preferentially sensitive to different rates of modulation are presented. The model supports the hypothesis that cells in the ICC decode temporal information into a rate code [Langner and Schreiner, J. Neurophysiol. 60, 1799-1822 (1988)], and provides a candidate wiring diagram of how this may be achieved.

Animals↗

The cochlear nuclei in two patients with Usher syndrome type I.

HYPOTHESIS: Does long-term sound deprivation lead to degeneration of the cochlear nuclei in two Usher type I patients? METHODS: The cochlear nuclei of these patients were morphometrically analyzed and compared with two age-matched controls. Routine autopsy of the brainstems was performed before the design of this study was known. During this procedure, the ventral cochlear nucleus (VCN) can easily be damaged. Five partially damaged VCN could nevertheless be analyzed for this study, including the right VCN of Usher patient 1 and both VCN of Usher patient 2. Using 15 microm thick serial paraffine sections of the cochlear nuclei, estimates of volume, neuronal densities, number of cells and mean cell diameter of the dorsal cochlear nucleus (DCN) and VCN were obtained. RESULTS: This study presents unique material of the cochlear nuclei in two patients with Usher syndrome type I. Data regarding volume and total cell number of the VCN are influenced by the absence of a part of the VCN. Results suggest a decrease in mean cell diameter of the VCN in Usher patients. Other parameters of the VCN and DCN, however, showed no major differences between Usher type I patients and controls. CONCLUSION: Only minor degenerative changes are apparent in the cochlear nuclei of two patients with Usher type I, who were deprived of acoustic stimuli since birth.

Aged↗

The long-term effect of bilateral inferior colliculus ablation on auditory brainstem response in awake cats.

In this study auditory brainstem responses (ABRs) were recorded repeatedly in three awake cats before and after bilateral aspiration of the inferior colliculi (ICs). All surgical procedures were performed under anaesthesia with Nembutal. Postoperative recordings were obtained 1 week, 3 months, 8 months and 13 months after bilateral aspiration of the ICs in order to study the long-term changes in ABRs. The P5 amplitude was markedly reduced only at 1 week after aspiration. P1 and P2 amplitudes were affected slightly at 3 months and 1 year, but those of P3 and P4 decreased significantly. After perfusion of these cats, histological examination of the brainstem revealed complete ablation of both ICs, neuronal loss in the nucleus of the lateral lemniscus and the superior olivary complex and preservation of the cochlear nucleus. Our results suggest that retrograde degeneration of neuronal cells of the brainstem, which project to the IC from the nucleus of the lateral lemniscus and superior olivary complex except the cochlear nucleus, affect the peak amplitudes of the ABR over the long-term.

Animals↗

The intra-aural muscle reflex in retrocochlear pathology: a model study in the rabbit.

An animal model for the evaluation of the acoustic stapedius reflex is described. The intra-aural muscle reflex in the rabbit can be recorded with a technique identical to that used in man. The validity and reproducibility of the measurements are good. Lesions in the reflex pathway were shown to be followed by changes in reflex properties. The type of alteration depended on the location of the damage. Lesions in the dorsal cochlear nucleus were not followed by reflex changes. Lesions in the ventral cochlear nucleus and/or the eighth nerve were followed by a rise of reflex threshold and often, but not always, by reflex decay. Lesions in the midline of the trapezoid body were followed by specific changes in the crossed reflex, whereas the ipsilateral reflex was unaffected. The advantage of making simultaneous ipsilateral and contraleral reflex recordings was pointed out.

Acoustic Stimulation↗

Expression of the c-fos transcription factor in the rat auditory pathway following postnatal auditory deprivation.

As an animal model for inborn hearing loss rat pups were reared in a sound-proof chamber from birth until age 21 days. In addition, pinnae were bilaterally sutured closed to reduce any influence of ambient sound. At the end of the sound deprivation, outer ear channels were reopened. Since previous studies failed to show any difference in the number or morphology of neurons in the auditory pathway in bilaterally sound-deprived animals, expression of c-fos protein was used as a functional marker to map trans-synaptic information transfer in the auditory pathway with cellular resolution. At day 21 sound-deprived rats and untreated controls were stimulated with pure tones of 8kHz for 5min at different sound pressure levels. Acoustic stimulation induced c-fos in both parts of the cochlear nucleus, superior olivary complex and inferior colliculus. Compared to untreated rats, deprivation reduced the number of c-fos labeled neurons in the dorsal and ventral part of the cochlear nucleus and inferior colliculus by 58% and 30%, respectively, following low sound pressure levels (90dB). In contrast, high sound pressure levels (120dB) increased the number of c-fos labeled neurons in these areas and evoked only minor differences in the number of labeled neurons in both untreated and sound deprived rats.

Acoustic Stimulation↗

Spatial organization of inner hair cell synapses and cochlear spiral ganglion neurons.

The morphological organization of the central projections of the cat cochlear spiral ganglion into the cochlear nucleus was previously investigated by creating restricted lesions in the anteroventral cochlear nucleus (AVCN) to ablate selectively either the lateral or the medial aspect of isofrequency projection laminae. Such lesions resulted in highly selective retrograde degeneration of spiral ganglion cells. Ablation of the lateral part of the AVCN caused degeneration of cells within the scala tympani part of the ganglion, whereas medial ablations within the AVCN induced degeneration of the scala vestibuli aspect of the ganglion. The peripheral axons also degenerated and this fiber loss exhibited selective topographies that paralleled the cell loss within the spiral ganglion, although this phenomenon was more prominent in the proximal part of the osseous spiral lamina near the ganglion and less obvious more distally near the habenula perforata. In this investigation, inner hair cells (IHCs) from these selective lesion cases were evaluated by electron microscopy of serial sections through the basal synaptic regions. Results demonstrated differential degeneration of afferent synapses, with greater (but not completely selective) loss of pillar synapses after lateral AVCN lesions and greater loss of modiolar synapses after medial lesions. Because auditory nerve fibers of different spontaneous discharge rates (SRs) have different spatial distributions on the IHC (Liberman, Science 216:1239, 1982), our results suggest that this SR-based organization is maintained in a topographic organization across the vertical (scala tympani-to-scala vestibuli) dimension of the spiral ganglion cell cluster and carried into the ventral cochlear nuclei (VCN). Thus, in addition to the spiral frequency organization represented by the dorsal-to-ventral frequency map in the VCN, there is also an orderly organization of inputs from high- and low-SR fibers across the lateral-to-medial dimension of the VCN such that the lateral isofrequency laminae receive a proportionately greater input from high-SR fibers, whereas medial isofrequency laminae receive preferential input from low- and medium-SR fibers.

Animals↗

Evidence for an auditory subsector within the zona incerta of rats.

This study explores the patterns of connections between the zona incerta (ZI) of the thalamus and the major auditory centres of the rat brain. Two series of experiments were performed using neuroanatomical tract tracing methods (biotinylated dextran or cholera toxin subunit b). First, tracers were injected into the ZI and resultant patterns of labelling in the auditory centres were examined. Labelling was seen in distinct subdivisions of the temporal cortex (Te3), medial geniculate complex (medial division; MGm), inferior colliculus (external cortex; ICe) and cochlear nucleus (dorsal and ventral divisions). For the most part, these subdivisions are involved in more associative or integrative aspects of audition. In a second series of experiments, three of these centres (Te3, ICe, cochlear nucleus) were injected with tracers and the labelling patterns formed in the ZI were explored. These results show that each of these connections makes a very distinct territory or subsector within the most lateral region of the ZI. This subsector of connectivity with the auditory centres does not respect the well defined cytoarchitectonic sectors of ZI, being made up of small slabs in both the dorsal and ventral sectors. Overall, the results suggest that ZI may integrate auditory information together with the other exteroceptive and interoceptive information that it receives and then influences subsequently global states of arousal and/or attention.

Animals↗

Effects of L-baclofen and D-baclofen on the auditory system: a study of click-evoked potentials from the inferior colliculus in the rat.

The drug baclofen is a potential treatment for severe tinnitus, but its action in relieving tinnitus is not known. Baclofen is available as an approved drug only in racemic form with about equal content of the two enantiomers. In the present paper we show that L-baclofen causes a considerable (40.7%) suppression of the amplitude of the second peak in the click-evoked response from the cochlear nucleus. Bipolar recordings from the external nucleus of the inferior colliculus showed that L-baclofen caused a reduction in the amplitude of three or four distinct peaks in this response. D-Baclofen had no detectable effect on the response from the cochlear nucleus, and had only a slight effect on one component of the response from the external nucleus of the inferior colliculus. The demonstrated effect of L-baclofen on excitation in the ascending auditory pathway indicates that this drug may be a potential treatment for hyperactive auditory disorders such as tinnitus and hyperacusis.

Animals↗

Role of acoustic striae in hearing: discrimination of sound-source elevation.

After years of systematic experimentation, we finally uncovered one thing the dorsal system contributes to hearing which the ventral system may not -- the mechanism for orienting to an elevated sound source [Sutherland, D.P., Masterton, R.B., Glendenning, K.K. (1998) Behav. Brain Res. in press]. This paper follows up this one positive result on a historical background of uniformly negative results. The focus of this report is on the fusiform cells of the dorsal cochlear nucleus whose axons course through the dorsal acoustic stria (DAS). Because electrophysiological studies have shown that the cues for sensing the elevation of a sound source would seem to be best analyzed by the dorsal cochlear nucleus, we tested, behaviorally, normal cats and cats deprived of their DAS or intermediate acoustic stria, bilaterally or ipsilaterally (with or without their contralateral ear deafened), for their ability to orient to elevated sources of broad-band noise. For behavioral testing, we made use of a conventional shock-avoidance procedure. The results lead to the conclusion that DCN and DAS may play no role in learned elevation discriminations. This result builds on that of another of our papers which suggests that a deficit in reflexive discrimination of elevation is strictly auditory in nature [Sutherland, D.P., Masterton, R.B., Glendenning, K.K. (1998) Behav. Brain Res. in press].

Acoustic Stimulation↗

Age-related changes in the distribution of Kv1.1 and Kv3.1 in rat cochlear nuclei.

OBJECTIVES: To identify age-related changes in voltage-gated K(+) (Kv) channels that contribute to temporal processing in neurons of the central auditory system, we investigated the distribution of Kv1.1 and Kv3.1 in the auditory brainstem of adult and aged rats. METHODS: Immunohistochemistry was performed in accordance with the free-floating method described earlier. RESULTS: Among the auditory nuclei, only the posterior ventral cochlear nucleus (PVCN) showed age-related changes. Kv1.1 immunoreactivity was increased in the octopus cell bodies, while the staining intensity was significantly decreased in the neuropil. Image analysis demonstrated the specific increase in Kv1.1 immunoreactivity in aged cochlear nucleus neurons although the mean density of the entire selection was significantly decreased. In contrast, the number of Kv1.1-immunoreactive neurons was not significantly different between control and aged groups. The immunoreactivity for Kv3.1 was decreased in the octopus cells and neuropil of aged PVCN, which was confirmed by image analysis. The number of Kv3.1-positive cells was also significantly decreased in aged PVCN. DISCUSSION: This study may provide useful data to compare age-related changes in Kv1.1 and Kv3.1 with known physiological properties of auditory neurons.

Age Factors↗

Development of brain-derived neurotrophic factor and neurotrophin-3 immunoreactivity in the lower auditory brainstem of the postnatal gerbil.

The localization of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) in the gerbil auditory brainstem was studied during normal postnatal development. The principal objective of this paper was to compare the developmental distribution of BDNF and NT-3 proteins to the known developmental distribution of their cognate, high-affinity tyrosine kinase receptors. BDNF and NT-3 proteins were localized using standard immunohistochemistry. No specific immunoreactivity for BDNF or NT-3 was detected on the day of birth (P0) in any auditory structure, although fibers comprising the spinal tract of the Vth cranial nerve were well labelled with antibodies against BDNF. Diffuse immunoreactivity for both BDNF and NT-3 was first detected at P3 in the cochlear nucleus and in several second order auditory nuclei in the superior olivary complex. This diffuse immunoreactivity became clustered and restricted to neuronal cell bodies by P10. Immunoreactivity for both BDNF and NT-3 transiently disappeared in the lateral and medial superior olivary nuclei at P10. However, neurons in the medial nucleus of the trapezoid body remained immunopositive for both BDNF and NT-3. Fibers in the trapezoid body were labelled with BDNF immunoreactivity by P12. Between P12 and P15, the distribution of BDNF and NT-3 immunoreactivity in the cochlear nucleus and superior olivary complex became comparable to adult (P140) immunolabel. These results show that the normal developmental distribution of the neurotrophins BDNF and NT-3 in the lower auditory brainstem occurs during the first two postnatal weeks in parallel with the developmental expression of their cognate receptors, trkB and trkC.

Animals↗

Distribution of mRNAs coding for liver and heart gap junction proteins in the rat central nervous system.

The present study examined the distributions of connexin43 mRNA and connexin32 mRNA in the central nervous system (CNS) of the rat by using in situ hybridization histochemistry. These connexins are the best studied gap junction proteins; connexin32 forms direct cell-cell channels in the liver, as does connexin43 in the heart. There was a differential distribution of cells containing connexin32 mRNA compared with the population of cells which contained connexin43 mRNA, thus implying a regional specificity in the expression of connexins in the CNS. Cells containing connexin43 mRNA were uniformly distributed throughout the gray matter of the neuraxis. Several areas had a higher concentration of cells that express connexin43, such as layer IA of the piriform cortex, supraoptic and paraventricular nuclei of the hypothalamus, anterior cortical amygdaloid nucleus, the reticular part of the substantia nigra, lateral habenula, mesencephalic trigeminal nucleus. Purkinje cell layer of the cerebellum, facial nucleus, prepositus hypoglossal nucleus, and dorsal cochlear nucleus. The pattern of connexin43 hybridization and the morphology of connexin43 mRNA containing cells suggest that this gap junction forming protein is found predominantly in astrocytes. Connexin32 mRNA was detected in discrete cell groups of the gray matter that appeared to be neurons, including cells in layer 2 of the neocortex, layer II of the piriform cortex, pyramidal cell layer of the hippocampus, granule and polymorphic cell layers of the dentate gyrus, islands of Calleja, olfactory tubercle, lateral thalamic nuclei, lateral habenula, and Purkinje cell layer of the cerebellar cortex. A large population of cells in white matter tracts that were labelled with the connexin32 riboprobe appeared to be oligodendrocytes. These studies suggest that neurons and glial cells express connexin32 mRNA, but only astrocytes express connexin43 mRNA. Many of the areas in which connexin mRNAs were demonstrated have electrically coupled cells, morphologically distinct gap junction plaques, and/or have immunocytochemically identifiable connexin proteins. These results indicate that cells with mRNAs coding for intercellular channels have a widespread distribution in the mammalian CNS.

Animals↗

An anatomical substrate for the inhibitory gradient in the VLVp of the owl.

The interaural difference in the level of sounds is an important cue for the localization of the sound's source. In the barn owl, a keen auditory predator, this binaural cue is first computed in the nucleus ventralis lemnisci laterale, pars posterior (VLVp), a cell group found within the fibers of the lateral lemniscus. Its neurons are excited by inputs from the contralateral ear and inhibited by inputs to the ipsilateral ear and are therefore sensitive indicators of interaural level difference. The excitation arrives by a direct input from the contralateral nucleus angularis, a cochlear nucleus, and the inhibition is mediated by a commissural projection that interconnects the VLVps of the two sides. The dorsally located neurons in the VLVp are more heavily inhibited than those found more ventrally, thus giving rise to a gradient of inhibition. This inhibitory gradient plays a central role in recent models of VLVp function. We present evidence based on standard anterograde tracing methods that this gradient of inhibition is mediated by a dorsoventral gradient in the density of synaptic inputs from the contralateral VLVp, the source of inhibition. Specifically, injection of tracers into one VLVp, regardless of the position of the injection within the nucleus, produced a vertically oriented field of label that was densest along the dorsal margin of the contralateral VLVp and became sparser a more ventral levels. Furthermore, we found that injections into the medial and lateral aspects of the nucleus produced this dorsoventrally graded field of label along the medial and lateral aspects of the contralateral VLVp, respectively. Finally, we confirmed an earlier observation suggesting that the anterior and posterior aspects of one VLVp project to the anterior and posterior aspects of the contralateral nucleus, respectively.

Animals↗

Comparative effects of pentylenetetrazol on the sensory responsiveness of lateral geniculate and reticular formation neurons.

The responses of bulbar and mesencephalic reticular formation (MRF) neurons to visual, auditory and/or somatosensory stimuli were considerably enhanced after subconvulsant doses of pentylenetetrazol (PTZ) in a similar fashion suggesting a general action of PTZ on reticular formation (RF) neurons. PTZ enhanced MRF responses evoked by electrical stimuli in the lateral geniculate nucleus (LGN) or cochlear nucleus but only modestly enhanced LGN neuronal responses. These findings indicate that the effects of this convulsant on the first brain sensory 'relay' nuclei and primary sensory receptors do not appear to be sufficient to account for the extensive PTZ-induced enhancement of RF neuronal responses, and direct effects of PTZ on the reticular formation may play a major role in this enhancement.

Animals↗