RESUMO
The large external pinnae and extensive vocal repertoire of the African wild dog (Lycaon pictus) has led to the assumption that the auditory system of this unique canid may be specialized. Here, using cytoarchitecture, myeloarchitecture, and a range of immunohistochemical stains, we describe the systems-level anatomy of the auditory system of the African wild dog. We observed the cochlear nuclear complex, superior olivary nuclear complex, lateral lemniscus, inferior colliculus, medial geniculate body, and auditory cortex all being in their expected locations, and exhibiting the standard subdivisions of this system. While located in the ectosylvian gyri, the auditory cortex includes several areas, resembling the parcellation observed in cats and ferrets, although not all of the auditory areas known from these species could be identified in the African wild dog. These observations suggest that, broadly speaking, the systems-level anatomy of the auditory system, and by extension the processing of auditory information, within the brain of the African wild dog closely resembles that observed in other carnivores. Our findings indicate that it is likely that the extraction of the semantic content of the vocalizations of African wild dogs, and the behaviors generated, occurs beyond the classically defined auditory system, in limbic or association neocortical regions involved in cognitive functions. Thus, to obtain a deeper understanding of how auditory stimuli are processed, and how communication is achieved, in the African wild dog compared to other canids, cortical regions beyond the primary sensory areas will need to be examined in detail.
Assuntos
Animais Selvagens/anatomia & histologia , Canidae/anatomia & histologia , Núcleo Coclear/anatomia & histologia , Nervo Vestibulococlear/anatomia & histologia , África , Animais , Córtex Auditivo , Vias Auditivas , Núcleo Coclear/fisiologia , Cães , Corpos Geniculados , Colículos Inferiores , Tegmento Pontino , Núcleos Talâmicos , Nervo Vestibulococlear/fisiologiaRESUMO
Located in the midbrain, the inferior colliculus (IC) is the hub of the central auditory system. Although the IC plays important roles in speech processing, sound localization, and other auditory computations, the organization of the IC microcircuitry remains largely unknown. Using a multifaceted approach in mice, we have identified vasoactive intestinal peptide (VIP) neurons as a novel class of IC principal neurons. VIP neurons are glutamatergic stellate cells with sustained firing patterns. Their extensive axons project to long-range targets including the auditory thalamus, auditory brainstem, superior colliculus, and periaqueductal gray. Using optogenetic circuit mapping, we found that VIP neurons integrate input from the contralateral IC and the dorsal cochlear nucleus. The dorsal cochlear nucleus also drove feedforward inhibition to VIP neurons, indicating that inhibitory circuits within the IC shape the temporal integration of ascending inputs. Thus, VIP neurons are well-positioned to influence auditory computations in a number of brain regions.
Assuntos
Colículos Inferiores/anatomia & histologia , Colículos Inferiores/fisiologia , Rede Nervosa/anatomia & histologia , Neurônios/química , Neurônios/fisiologia , Peptídeo Intestinal Vasoativo/análise , Animais , Núcleo Coclear/anatomia & histologia , Camundongos , Técnicas de Rastreamento Neuroanatômico , Neurônios/classificação , OptogenéticaRESUMO
Hearing depends on extracting frequency, intensity, and temporal properties from sound to generate an auditory map for acoustical signal processing. How physiology intersects with molecular specification to fine tune the developing properties of the auditory system that enable these aspects remains unclear. We made a novel conditional deletion model that eliminates the transcription factor NEUROD1 exclusively in the ear. These mice (both sexes) develop a truncated frequency range with no neuroanatomically recognizable mapping of spiral ganglion neurons onto distinct locations in the cochlea nor a cochleotopic map presenting topographically discrete projections to the cochlear nuclei. The disorganized primary cochleotopic map alters tuning properties of the inferior colliculus units, which display abnormal frequency, intensity, and temporal sound coding. At the behavioral level, animals show alterations in the acoustic startle response, consistent with altered neuroanatomical and physiological properties. We demonstrate that absence of the primary afferent topology during embryonic development leads to dysfunctional tonotopy of the auditory system. Such effects have never been investigated in other sensory systems because of the lack of comparable single gene mutation models.SIGNIFICANCE STATEMENT All sensory systems form a topographical map of neuronal projections from peripheral sensory organs to the brain. Neuronal projections in the auditory pathway are cochleotopically organized, providing a tonotopic map of sound frequencies. Primary sensory maps typically arise by molecular cues, requiring physiological refinements. Past work has demonstrated physiologic plasticity in many senses without ever molecularly undoing the specific mapping of an entire primary sensory projection. We genetically manipulated primary auditory neurons to generate a scrambled cochleotopic projection. Eliminating tonotopic representation to auditory nuclei demonstrates the inability of physiological processes to restore a tonotopic presentation of sound in the midbrain. Our data provide the first insights into the limits of physiology-mediated brainstem plasticity during the development of the auditory system.
Assuntos
Percepção Auditiva/genética , Percepção Auditiva/fisiologia , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/fisiologia , Mesencéfalo/fisiologia , Percepção da Altura Sonora/fisiologia , Animais , Comportamento Animal/fisiologia , Mapeamento Encefálico , Núcleo Coclear/anatomia & histologia , Núcleo Coclear/fisiologia , Feminino , Audição/fisiologia , Colículos Inferiores/anatomia & histologia , Colículos Inferiores/fisiologia , Masculino , Mesencéfalo/embriologia , Camundongos , Camundongos Knockout , Gravidez , Reflexo de Sobressalto/genética , Reflexo de Sobressalto/fisiologia , Gânglio Espiral da Cóclea/citologia , Gânglio Espiral da Cóclea/fisiologia , Vestíbulo do Labirinto/anatomia & histologia , Vestíbulo do Labirinto/fisiologiaRESUMO
The human auditory brainstem, especially the cochlear nucleus (CN) and the superior olivary complex (SOC) are characterized by a high density of neurons associated with perineuronal nets (PNs). PNs build a specific form of extracellular matrix surrounding the neuronal somata, proximal dendrites and axon initial segments. They restrict synaptic plasticity and control high-frequency synaptic activity, a prominent characteristic of neurons of the auditory brainstem. The distribution of PNs within the auditory brainstem has been investigated in a number of mammalian species. However, much less is known regarding PNs in the human auditory brainstem. The present study aimed at the immunohistochemical identification of PNs in the cochlear nucleus (CN) and superior olivary complex (SOC) in the human brainstem. We focused on the complex nature and molecular variability of PNs in the CN and SOC by using specific antibodies against the main PN components (aggrecan, brevican, neurocan and hyaluronan and proteoglycan link protein 1). Virtually all subnuclei within the ventral CN and SOC were found to be associated with PNs. Direct comparison between gerbil and human yielded similar fine structure of PNs and confirmed the typical tight interdigitation of PNs with synaptic terminals in both species. Noticeably, an elaborate combination of immunohistochemical labelings clearly supports the still debated existence of the medial nucleus of trapezoid body (MNTB) in the human brain. In conclusion, the present study demonstrates that PNs form a prominent extracellular structure on CN and SOC neurons in the human brain, potentially stabilizing synaptic contacts, which is in agreement with many other mammalian species.
Assuntos
Vias Auditivas/anatomia & histologia , Núcleo Coclear/anatomia & histologia , Rede Nervosa/anatomia & histologia , Terminações Pré-Sinápticas , Complexo Olivar Superior/anatomia & histologia , Idoso de 80 Anos ou mais , Agrecanas/análise , Animais , Vias Auditivas/química , Biomarcadores/análise , Brevicam/análise , Cadáver , Proteoglicanas de Sulfatos de Condroitina/análise , Núcleo Coclear/química , Feminino , Gerbillinae , Humanos , Ácido Hialurônico/análise , Imuno-Histoquímica , Lectinas Tipo C/análise , Masculino , Pessoa de Meia-Idade , Rede Nervosa/química , Proteínas do Tecido Nervoso/análise , Técnicas de Rastreamento Neuroanatômico , Neurocam , Terminações Pré-Sinápticas/química , Complexo Olivar Superior/química , Corpo Trapezoide/anatomia & histologia , Corpo Trapezoide/químicaRESUMO
Auditory impulses perceived by the hair cells of the organ of corti are relayed in the cochlear nucleus, the first relay station in the brainstem, by the cochlear nerve. The human foetus is well known to respond to sound during the last trimester of gestation. On the contrary, studies conducted in rat, cat and mouse have shown that these mammals have an immature auditory system at the time of birth. There are very few reports available regarding the morphological and functional maturation of the cochlear nucleus in human. Although the human cochlear nucleus neurons attain adult morphological characters by mid-gestation, there are hardly any studies discussing the functional maturation of the cochlear nucleus. Hence the present study was aimed at observing the morphological as well as functional maturation of the human foetal cochlear nuclei at various gestational ages. Morphological maturation was observed qualitatively while stereological estimation of the volume of well defined ventral cochlear nucleus (VCN) was calculated by the Cavalieri principle; neuronal count and density was estimated by dissector principle. The functional maturation was assessed by observing the expression of synaptophysin, a synaptic marker, at different gestational ages and by the presence of parvalbumin, a calcium binding functional neuronal marker by immunohistochemistry. Neurons showed coarse Nissl's substance and well developed cell processes and gradual increase in cell size by the 24th-30th gestational week. Synaptophysin labeling in the complete cochlear nucleus was observed at 20 weeks of gestation. Adult pattern of synaptophysin labeling was observed finally at37weeks of gestation. Earliest presence of parvalbumin expression was detected at 16 weeks of gestation and a distinct adult pattern was seen at 37 weeks of gestation. This study concluded that morphological and functional maturation of the human cochlear nuclei occurs simultaneously during mid-gestation which represents the critical period of development and continues up to term.
Assuntos
Núcleo Coclear/anatomia & histologia , Núcleo Coclear/embriologia , Adulto , Vias Auditivas , Contagem de Células , Tamanho Celular , Núcleo Coclear/metabolismo , Feminino , Idade Gestacional , Humanos , Imageamento Tridimensional , Imuno-Histoquímica , Masculino , Neurônios/ultraestrutura , Corpos de Nissl/ultraestrutura , Parvalbuminas/metabolismo , Gravidez , Sinaptofisina/metabolismoRESUMO
Ascending projections of the dorsal cochlear nucleus (DCN) target primarily the contralateral inferior colliculus (IC). In turn, the IC sends bilateral descending projections back to the DCN. We sought to determine the nature of these descending axons in order to infer circuit mechanisms of signal processing at one of the earliest stages of the central auditory pathway. An anterograde tracer was injected in the IC of CBA/Ca mice to reveal terminal characteristics of the descending axons. Retrograde tracer deposits were made in the DCN of CBA/Ca and transgenic GAD67-EGFP mice to investigate the cells giving rise to these projections. A multiunit best frequency was determined for each injection site. Brains were processed by using standard histologic methods for visualization and examined by fluorescent, brightfield, and electron microscopy. Descending projections from the IC were inferred to be excitatory because the cell bodies of retrogradely labeled neurons did not colabel with EGFP expression in neurons of GAD67-EGFP mice. Furthermore, additional experiments yielded no glycinergic or cholinergic positive cells in the IC, and descending projections to the DCN were colabeled with antibodies against VGluT2, a glutamate transporter. Anterogradely labeled endings in the DCN formed asymmetric postsynaptic densities, a feature of excitatory neurotransmission. These descending projections to the DCN from the IC were topographic and suggest a feedback pathway that could underlie a frequency-specific enhancement of some acoustic signals and suppression of others. The involvement of this IC-DCN circuit is especially noteworthy when considering the gating of ascending signal streams for auditory processing. J. Comp. Neurol. 525:773-793, 2017. © 2016 Wiley Periodicals, Inc.
Assuntos
Núcleo Coclear/fisiologia , Colículos Inferiores/fisiologia , Animais , Vias Auditivas/anatomia & histologia , Vias Auditivas/fisiologia , Núcleo Coclear/anatomia & histologia , Eletrofisiologia , Imunofluorescência , Colículos Inferiores/anatomia & histologia , CamundongosRESUMO
Axons of olivocochlear neurons originate from the brainstem and project to the cochlea. A subpopulation, medial olivocochlear (MOC) neurons, also projects collateral branches to the cochlear nucleus. The precise targets of these collaterals are as yet unknown. Previous methods for labelling these collaterals include firstly, cochlear injections of retrograde tracers, but this is technically demanding and can also label afferent projections or secondly, labelling by injecting tracers into the nuclei of origin of MOC neurons. However, this latter method is non-specific because it also labels non-MOC projections. A technique was used to specifically label MOC collaterals, which involved injections of the tracer biocytin at the floor of the fourth ventricle and fixation 3 hours later. Biocytin injections resulted in labelled neurons in the ventral nucleus of the trapezoid body and rostral periolivary nucleus, confirming MOC axonal labelling. Labelled neurons in dorsal cochlear nucleus indicated labelling of the dorsal acoustic stria and these injections were discarded. After selective MOC labelling, collateral branches were found to innervate granule cell regions, medial edge and core of the ventral cochlear nucleus, as well as the dorsal cochlear nucleus, in agreement with previous data. Therefore we conclude that injections at the floor of the fourth ventricle provide a simple, rapid and specific technique for labelling the majority of MOC axons and their collaterals and this technique may assist in defining the precise neuronal targets of olivocochlear collaterals in cochlear nucleus.
Assuntos
Vias Auditivas/fisiologia , Cóclea/inervação , Núcleo Coclear/fisiologia , Núcleo Olivar/fisiologia , Acústica , Animais , Axônios/fisiologia , Tronco Encefálico/fisiologia , Núcleo Coclear/anatomia & histologia , Feminino , Quarto Ventrículo/patologia , Lisina/análogos & derivados , Lisina/química , Masculino , Modelos Neurológicos , Neurônios/metabolismo , Ratos , Ratos Wistar , Núcleos Vestibulares/fisiologiaRESUMO
BACKGROUND: Auditory brainstem implantation at the cochlear nuclei used mainly for neurofibromatosis type 2 patients with bilateral loss of the cochlear nerves has more recently been extended to the inferior colliculus. OBJECTIVE: To examine the microsurgical and endoscopic anatomy of the cochlear nuclei and inferior colliculus as seen through the translabyrinthine and retrosigmoid approaches used for cochlear nuclei and inferior collicular implantation. METHODS: Ten cerebellopontine angles of formalin-fixed adult cadaveric heads were examined with the aid of the surgical microscope and endoscope. The ascending auditory pathways between the cochlear nuclei and inferior colliculi and above were examined by the fiber dissection technique. RESULTS: Both the translabyrinthine and retrosigmoid routes provide sufficient exposure for concurrent tumor removal and implantation at either the cochlear nuclei or inferior colliculus. The position of the inferior colliculus in the auditory pathways and its accessibility in the infratentorial supracerebellar exposure directed through either the translabyrinthine or retrosigmoid approach makes it an alternative site for electrode placement if the cochlear nuclei are not functionally or structurally suitable for implantation. Endoscopic assistance may aid the exposure and electrode placement at either site. CONCLUSION: The translabyrinthine or retrosigmoid approaches provide access to the cochlear nuclei for implantation and also to the inferior colliculus through the translabyrinthine or retrosigmoid infratentorial supracerebellar route. The endoscope may aid in exposing either site.
Assuntos
Implante Auditivo de Tronco Encefálico/métodos , Núcleo Coclear/anatomia & histologia , Colículos Inferiores/anatomia & histologia , Adulto , Cadáver , Ângulo Cerebelopontino/cirurgia , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Auditory brainstem implants have failed to produce consistent clinical results comparable to those with the cochlear implant, both with surface and penetrating electrodes. OBJECTIVE: To determine neuromorphological constraints of the auditory brainstem implant interface. METHODS: The size, shape, surface depth, and spatial orientation of 33 human cochlear nuclei in 20 brainstem specimens obtained at autopsy were systematically analyzed in 792 slices each with a thickness of 8 µm. Three-dimensional renderings of the cochlear nucleus complex were obtained from a true-to-scale model, and the resulting photographic views were arranged according to the axes of the brainstem. RESULTS: The dimensions of the ventral and dorsal cochlear nuclei in the axial, coronal, and sagittal planes correlated linearly with each other. There were no significant side differences. Maximum dimensions of the whole cochlear nuclear complex were 8.01 × 1.53 × 3.76 mm. The appearance of the ventral and dorsal nuclei combined resembles a distorted X shape from a lateral view and an angulated wedge shape when viewed from above. Slanted into the depth of the brainstem above the facial nerve entrance, the superior boundary of the ventral nucleus is located more than 7 mm off the surface of the brainstem on average. CONCLUSION: In the absence of appropriate surface landmarks and imaging guidance, to gain tonotopic access to the human cochlear nucleus with surface and depth electrode remains a major challenge. Due to its location close to the surface, the dorsal cochlear nucleus is vulnerable to surgical manipulation and to tumors.
Assuntos
Implantes Auditivos de Tronco Encefálico , Núcleo Coclear/anatomia & histologia , Idoso , Idoso de 80 Anos ou mais , Cadáver , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The relationship between structure and function is an invaluable context with which to explore biological mechanisms of normal and dysfunctional hearing. The systematic and topographic representation of frequency originates at the cochlea, and is retained throughout much of the central auditory system. The cochlear nucleus (CN), which initiates all ascending auditory pathways, represents an essential link for understanding frequency organization. A model of the CN that maps frequency representation in 3D would facilitate investigations of possible frequency specializations and pathologic changes that disturb frequency organization. Toward this goal, we reconstructed in 3D the trajectories of labeled auditory nerve (AN) fibers following multiunit recordings and dye injections in the anteroventral CN of the CBA/J mouse. We observed that each injection produced a continuous sheet of labeled AN fibers. Individual cases were normalized to a template using 3D alignment procedures that revealed a systematic and tonotopic arrangement of AN fibers in each subdivision with a clear indication of isofrequency laminae. The combined dataset was used to mathematically derive a 3D quantitative map of frequency organization throughout the entire volume of the CN. This model, available online (http://3D.ryugolab.com/), can serve as a tool for quantitatively testing hypotheses concerning frequency and location in the CN.
Assuntos
Núcleo Coclear/anatomia & histologia , Imageamento Tridimensional , Estimulação Acústica , Animais , Nervo Coclear/anatomia & histologia , Nervo Coclear/fisiologia , Núcleo Coclear/fisiologia , Eletrofisiologia , Feminino , Masculino , Camundongos , Camundongos Endogâmicos CBARESUMO
The axons of commissural neurons that project from one cochlear nucleus to the other were studied after labeling with anterograde tracer. Injections were made into the dorsal subdivision of the cochlear nucleus in order to restrict labeling only to the group of commissural neurons that gave off collaterals to, or were located in, this subdivision. The number of labeled commissural axons in each injection was correlated with the number of labeled radiate multipolar neurons, suggesting radiate neurons as the predominant origin of the axons. The radiate commissural axons are thick and myelinated, and they exit the dorsal acoustic stria of the injected cochlear nucleus to cross the brainstem in the dorsal half, near the crossing position of the olivocochlear bundle. They enter the opposite cochlear nucleus via the dorsal and ventral acoustic stria and at its medial border. Reconstructions of single axons demonstrate that terminations are mostly in the core and typically within a single subdivision of the cochlear nucleus. Extents of termination range from narrow to broad along both the dorsoventral (i.e., tonotopic) and the rostrocaudal dimensions. In the electron microscope, labeled swellings form synapses that are symmetric (in that there is little postsynaptic density), a characteristic of inhibitory synapses. Our labeled axons do not appear to include excitatory commissural axons that end in edge regions of the nucleus. Radiate commissural axons could mediate the broadband inhibition observed in responses to contralateral sound, and they may balance input from the two ears with a quick time course.
Assuntos
Vias Auditivas/anatomia & histologia , Núcleo Coclear/anatomia & histologia , Animais , Axônios , Feminino , Masculino , Camundongos , Camundongos Endogâmicos CBARESUMO
HYPOTHESIS: Optimal pharmacotherapy entails a safe delivery method that specifically targets auditory structure(s) of interest. A retrocochlear neuronal tracer may enable comparison of various pharmacotherapy delivery methods and localization of the drug along the auditory pathway. BACKGROUND: Sensorineural hearing loss (SNHL) can involve cochlear hair cell or neural cell death, which often is accompanied by secondary degeneration of central auditory neurons. Targeting the precise location of nerve degeneration is important for treatment success. To be clinically relevant, the method of drug delivery must be safe and reliable while being maximally absorbed by the relevant inner ear structures of interest. METHODS: We compared 3 methods of FluoroGold (FG) delivery, a retrograde neuronal tracer, in delineating the retrocochlear auditory pathway using a normal-hearing strain of CBA mice. FG was delivered either intratympanic (IT), intracochlear (IC), or through the round window (RW). Five days after FG injection, mice were sacrificed for cell counts in the cochlear nucleus (CN), superior olivary complex (SOC), and the lateral lemniscus (LL). RESULTS: Although neurons in the CN and SOC were abundantly labeled by FG in all 3 injection methods, the IT method was the most reproducible and specific. The average cells for the CN, SOC, and LL were 851 ± 121, 2629 ± 367, and 112 ± 30, respectively. Accurate cell counts could not be established for the IC and RW injection methods because of nonspecific cell staining. Only 1 of the 5 IC-injected mice had specific labeling along the retrocochlear auditory pathway. Cell counts for the single mouse with specific IC staining in the CN, SOC, and LL were 177, 1839, and 56, respectively. Similarly, 2 of the 5 RW-injected mice had specific labeling, whereas the rest were nonspecific. The average cell counts for the 2 mice with specific labeling in the CN, SOC, and LL was 723.5 ± 580.0, 2173.5 ± 998.0, and 131.5 ± 8.0, respectively. CONCLUSION: The IT injection method resulted in reproducible, specific staining of neuronal cells along the retrocochlear auditory pathway compared with the RW or IC route of delivery.
Assuntos
Vias Auditivas/anatomia & histologia , Nervo Coclear/anatomia & histologia , Sistemas de Liberação de Medicamentos , Perda Auditiva Neurossensorial/tratamento farmacológico , Marcadores do Trato Nervoso , Animais , Contagem de Células , Núcleo Coclear/anatomia & histologia , Orelha Interna/anatomia & histologia , Corantes Fluorescentes , Camundongos , Camundongos Endogâmicos CBA , Núcleo Olivar/anatomia & histologia , Reprodutibilidade dos Testes , Janela da Cóclea/anatomia & histologia , Estilbamidinas , Membrana Timpânica/anatomia & histologiaRESUMO
Despite the outstanding auditory capabilities of dolphins, there is only limited information available on the cytology of the auditory brain stem nuclei in these animals. Here, we investigated the cochlear nuclei (CN) of five brains of common dolphins (Delphinus delphis) and La Plata dolphins (Pontoporia blainvillei) using cell and fiber stain microslide series representing the three main anatomical planes. In general, the CN in dolphins comprise the same set of subnuclei as in other mammals. However, the volume ratio of the dorsal cochlear nucleus (DCN) in relation to the ventral cochlear nucleus (VCN) of dolphins represents a minimum among the mammals examined so far. Because, for example, in cats the DCN is necessary for reflexive orientation of the head and pinnae towards a sound source, the massive restrictions in head movability in dolphins and the absence of outer ears may be correlated with the reduction of the DCN. Moreover, the same set of main neuron types were found in the dolphin CN as in other mammals, including octopus and multipolar cells. Because the latter two types of neurons are thought to be involved in the recognition of complex sounds, including speech, we suggest that, in dolphins, they may be involved in the processing of their communication signals. Comparison of the toothed whale species studied here revealed that large spherical cells were present in the La Plata dolphin but absent in the common dolphin. These neurons are known to be engaged in the processing of low-frequency sounds in terrestrial mammals. Accordingly, in the common dolphin, the absence of large spherical cells seems to be correlated with a shift of its auditory spectrum into the high-frequency range above 20 kHz. The existence of large spherical cells in the VCN of the La Plata dolphin, however, is enigmatic asthis species uses frequencies around 130 kHz.
Assuntos
Núcleo Coclear/anatomia & histologia , Núcleo Coclear/fisiologia , Golfinhos Comuns/anatomia & histologia , Golfinhos Comuns/fisiologia , Animais , Encéfalo/anatomia & histologia , Encéfalo/fisiologia , Mapeamento Encefálico , Tronco Encefálico/anatomia & histologia , Tronco Encefálico/fisiologia , Neurônios/fisiologia , Som , Coloração e Rotulagem , Núcleos Vestibulares/anatomia & histologiaRESUMO
OBJECTIVE: To find morphological evidence of a direct projection from the cochlear nucleus (CN; at the brainstem level) in the auditory system to the peripheral end organs in the vestibular system. METHODS: Experiments were conducted on male Wistar rats (n = 24). Two neuronal tracers were used: (1) 5% molecular probe F-8793 was injected into the unilateral peripheral vestibule and used as a retrograde tracer; (2) PHA-L (Invitrogen L-11270) was injected into the unilateral CN and used as an anterograde tracer. All animals were allowed to recover for 7 days after surgery to facilitate sufficient transportation of the tracers. Subsequently, brainstems in the retrograde group and inner ears in the anterograde group were sliced coronally on a freezing microtome and observed under a fluorescence microscope. RESULTS: After PHA-L injection into the CN, terminal labeling was observed in the peripheral vestibule, especially in the inferior vestibular nerve. The retrograde tracing study showed that the positive cells could be found in the ventral part of the CN. CONCLUSIONS: These results suggest that there is a novel pathway with a consanguineous functional connection between the CN and peripheral vestibule.
Assuntos
Vias Auditivas/anatomia & histologia , Tronco Encefálico/anatomia & histologia , Nervo Coclear/anatomia & histologia , Núcleo Coclear/anatomia & histologia , Audição/fisiologia , Potenciais Evocados Miogênicos Vestibulares/fisiologia , Vestíbulo do Labirinto/anatomia & histologia , Animais , Vias Auditivas/fisiologia , Tronco Encefálico/fisiologia , Nervo Coclear/fisiologia , Núcleo Coclear/fisiologia , Masculino , Ratos , Ratos Wistar , Vestíbulo do Labirinto/fisiologiaRESUMO
Fast, temporally-precise, and consistent synaptic transmission is required to encode features of acoustic stimuli. Neurons of nucleus magnocellularis (NM) in the auditory brain stem of the chick possess numerous adaptations to optimize the coding of temporal information. One potential problem for the system is the depression of synaptic transmission during a prolonged stimulus. The present study tested the hypothesis that cannabinoid receptor 1 (CB1) signaling may limit synaptic depression at the auditory nerve-NM synapse. In situ hybridization was used to confirm that CB1 mRNA is expressed in the cochlear ganglion; immunohistochemistry was used to confirm the presence of CB1 protein in NM. These findings are consistent with the common presynaptic locus of CB1 in the brain. Rate-dependent synaptic depression was then examined in a brain slice preparation before and after administration of WIN 55,212-2 (WIN), a potent CB1 agonist. WIN decreased the amplitude of excitatory postsynaptic currents (EPSCs) and also reduced depression across a train of stimuli. The effect was most obvious late in the pulse train and during high rates of stimulation. This CB1-mediated influence could allow for lower, but more consistent activation of NM neurons, which could be of importance for optimizing the coding of prolonged, temporally-locked acoustic stimuli.
Assuntos
Vias Auditivas/fisiologia , Tronco Encefálico/fisiologia , Núcleo Coclear/fisiologia , Receptor CB1 de Canabinoide/metabolismo , Gânglio Espiral da Cóclea/fisiologia , Animais , Vias Auditivas/anatomia & histologia , Tronco Encefálico/anatomia & histologia , Embrião de Galinha , Galinhas , Núcleo Coclear/anatomia & histologia , Feminino , Masculino , Técnicas de Cultura de Órgãos , Receptor CB1 de Canabinoide/genética , Receptor CB1 de Canabinoide/fisiologia , Transdução de Sinais/fisiologia , Gânglio Espiral da Cóclea/anatomia & histologia , Sinapses/fisiologiaRESUMO
The chicken auditory brainstem is a well-established model system that has been widely used to study the anatomy and physiology of auditory processing at discreet periods of development as well as mechanisms for temporal coding in the central nervous system. Here we present a method to prepare chicken auditory brainstem slices that can be used for acute experimental procedures or to culture organotypic slices for long-term experimental manipulations. The chicken auditory brainstem is composed of nucleus angularis, magnocellularis, laminaris and superior olive. These nuclei are responsible for binaural sound processing and single coronal slice preparations preserve the entire circuitry. Ultimately, organotypic slice cultures can provide the opportunity to manipulate several developmental parameters such as synaptic activity, expression of pre and postsynaptic components, expression of aspects controlling excitability and differential gene expression This approach can be used to broaden general knowledge about neural circuit development, refinement and maturation.
Assuntos
Tronco Encefálico/anatomia & histologia , Núcleo Coclear/anatomia & histologia , Microtomia/métodos , Técnicas de Cultura de Órgãos/métodos , Animais , Tronco Encefálico/citologia , Embrião de Galinha , Núcleo Coclear/citologiaRESUMO
Acetylcholine (Ach) affects a variety of cell types in the cochlear nucleus (CN) and is likely to play a role in numerous functions. Previous work in rats suggested that the acetylcholine arises from cells in the superior olivary complex, including cells that have axonal branches that innervate both the CN and the cochlea (i.e. olivocochlear cells) as well as cells that innervate only the CN. We combined retrograde tracing with immunohistochemistry for choline acetyltransferase to identify the source of ACh in the CN of guinea pigs. The results confirm a projection from cholinergic cells in the superior olivary complex to the CN. In addition, we identified a substantial number of cholinergic cells in the pedunculopontine tegmental nucleus (PPT) and the laterodorsal tegmental nucleus (LDT) that project to the CN. On average, the PPT and LDT together contained about 26% of the cholinergic cells that project to CN, whereas the superior olivary complex contained about 74%. A small number of additional cholinergic cells were located in other areas, including the parabrachial nuclei.The results highlight a substantial cholinergic projection from the pontomesencephalic tegmentum (PPT and LDT) in addition to a larger projection from the superior olivary complex. These different sources of cholinergic projections to the CN are likely to serve different functions. Projections from the superior olivary complex are likely to serve a feedback role, and may be closely tied to olivocochlear functions. Projections from the pontomesencephalic tegmentum may play a role in such things as arousal and sensory gating. Projections from each of these areas, and perhaps even the smaller sources of cholinergic inputs, may be important in conditions such as tinnitus as well as in normal acoustic processing.
Assuntos
Vias Auditivas/citologia , Fibras Colinérgicas , Núcleo Coclear/anatomia & histologia , Acetilcolina/metabolismo , Animais , Vias Auditivas/metabolismo , Núcleo Coclear/metabolismo , Cobaias , Imuno-HistoquímicaRESUMO
The vesicular glutamate transporters (VGLUTs) regulate the storage and release of glutamate in the brain. In adult animals, the VGLUT1 and VGLUT2 isoforms are widely expressed and differentially distributed, suggesting that neural circuits exhibit distinct modes of glutamate regulation. Studies in rodents suggest that VGLUT1 and VGLUT2 mRNA expression patterns are partly complementary, with VGLUT1 expressed at higher levels in the cortex and VGLUT2 prominent subcortically, but with overlapping distributions in some nuclei. In primates, VGLUT gene expression has not been previously studied in any part of the brain. The purposes of the present study were to document the regional expression of VGLUT1 and VGLUT2 mRNA in the auditory pathway through A1 in the cortex, and to determine whether their distributions are comparable to rodents. In situ hybridization with antisense riboprobes revealed that VGLUT2 was strongly expressed by neurons in the cerebellum and most major auditory nuclei, including the dorsal and ventral cochlear nuclei, medial and lateral superior olivary nuclei, central nucleus of the inferior colliculus, sagulum, and all divisions of the medial geniculate. VGLUT1 was densely expressed in the hippocampus and ventral cochlear nuclei, and at reduced levels in other auditory nuclei. In the auditory cortex, neurons expressing VGLUT1 were widely distributed in layers II-VI of the core, belt and parabelt regions. VGLUT2 was expressed most strongly by neurons in layers IIIb and IV, weakly by neurons in layers II-IIIa, and at very low levels in layers V-VI. The findings indicate that VGLUT2 is strongly expressed by neurons at all levels of the subcortical auditory pathway, and by neurons in the middle layers of the cortex, whereas VGLUT1 is strongly expressed by most if not all glutamatergic neurons in the auditory cortex and at variable levels among auditory subcortical nuclei. These patterns imply that VGLUT2 is the main vesicular glutamate transporter in subcortical and thalamocortical (TC) circuits, whereas VGLUT1 is dominant in corticocortical (CC) and corticothalamic (CT) systems of projections. The results also suggest that VGLUT mRNA expression patterns in primates are similar to rodents, and establish a baseline for detailed studies of these transporters in selected circuits of the auditory system.
Assuntos
Córtex Auditivo/fisiologia , Regulação da Expressão Gênica , Proteína Vesicular 1 de Transporte de Glutamato/biossíntese , Proteína Vesicular 2 de Transporte de Glutamato/biossíntese , Animais , Aotus trivirgatus , Córtex Auditivo/anatomia & histologia , Mapeamento Encefálico/métodos , Núcleo Celular/metabolismo , Núcleo Coclear/anatomia & histologia , Corpos Geniculados/anatomia & histologia , Hibridização In Situ , Colículos Inferiores/anatomia & histologia , Núcleo Olivar/anatomia & histologia , Primatas , Distribuição Tecidual , Proteína Vesicular 1 de Transporte de Glutamato/genética , Proteína Vesicular 2 de Transporte de Glutamato/genéticaRESUMO
A major limitation of the present auditory brainstem implant (ABI) is its inability to access the tonotopic organization of the ventral cochlear nucleus (VCN). A previous study by our group indicated that stimulation of single sites within a given VCN frequency region did not always elicit frequency-specific responses within the central nucleus of the inferior colliculus (CIC) and in some cases did not elicit a response at all. For this study, we hypothesized that sequential stimulation (with a short interpulse delay of 320 µsec) of two VCN sites in similar frequency regions would enhance responsiveness in CIC neurons. Multiunit neural recordings in response to pure tones were obtained at 58 VCN and 164 CIC sites in anesthetized rats. Among the 58 VCN sites, 39 pairs of sites with similar characteristic frequencies were chosen for electrical stimulation. Each member of a VCN pair was electrically stimulated individually, followed by sequential stimulation of the pair, while recording CIC responses. On average, CIC sites were found to respond to dual-site VCN stimulation with significantly lower thresholds, wider dynamic ranges, a greater extent of activation with increasing current levels, and a higher degree of frequency specificity compared with single-site stimulation. Although these effects were positive for the most part, in some cases dual-site stimulation resulted in increased CIC thresholds and decreased dynamic ranges, extent of activation, and frequency specificity. The results suggest that multisite stimulation within VCN isofrequency laminae using penetrating electrodes could significantly improve ABI stimulation strategies and implant performance.