<p>The superior olivary complex (SOC) is comprised of nuclei involved in sound localization. To compute interaural sound level differences, lateral superior olive (LSO) neurons integrate converging glutamatergic inputs from the cochlear nucleus with glycinergic inputs from the medial nucleus of the trapezoid body (MNTB). To compute interaural timing differences, the medial superior olive (MSO) integrates converging glutamatergic inputs from the ipsilateral and contralateral cochlear nucleus. The MSO also receives a major inhibitory input from the MNTB. N-methyl-D-aspartate receptors (NMDARs) are thought to play a role in the developmental refinement of these auditory brainstem pathways. The GluN2A and GluN2B NMDAR subunits confer widely different properties on NMDARs, substantially affecting plasticity. We assessed postnatal developmental messenger RNA (mRNA) expression of GluN1, GluN2A and GluN2B subunits in the LSO, MSO and MNTB using quantitative <em>in-situ</em> hybridization in tissue from 10 litters, ages postnatal day 1 to 36 (P1-36).</p> <p>GluN1 mRNA expression in the LSO, MSO and MNTB decreased with age. In all three nuclei, GluN2B mRNA expression was highest during the first postnatal week, dropping to low levels thereafter. In the LSO, GluN2A levels increased, then decreased to moderate levels. In the MNTB, GluN2A levels decreased from initially high levels. In the MSO, GluN2A levels increased to intermediate levels. The GluN2A/2B ratio increased 2-fold between P1 and P8 in the MNTB, whereas the ratio increased 3-fold between P8 and P15 in the LSO and MSO. The changes in GluN2A:GluN2B ratio are consistent with a developmental switch from GluN2B-containing NMDARs to GluN2A-containing NMDARs. These results are consistent with prior electrophysiological experiments that show NMDAR-mediated currents declining with age in the aVCN-MNTB, aVCN-LSO and MNTB-LSO pathways. The GluN2A subunit exhibited different developmental expression patterns in MNTB, LSO and MNTB, which suggests that GluN2A mRNA expression is locally regulated between nuclei, whereas GluN2B may be globally regulated.</p> / Master of Science (MSc)

<p>The neural code from the inner ear to the brain is dynamically controlled by central nervous efferent feedback to the audio-vestibular epithelium. Although such efference provides the basis for a cognitive control of our hearing and balance, we know surprisingly little about this feedback system. This project has investigated the applicability of a transgenic mouse model, expressing a fluorescent protein under the choline-acetyltransferase (ChAT) promoter, for targeting the cholinergic audio-vestibular efferent neurons in the brainstem. It was found that the mouse model is useful for targeting the vestibular efferents, which are fluorescent, but not the auditory efferents, which are not highlighted. This model enables, for the first time, physiological studies of the vestibular efferent neurons and their synaptic inputs. We next assessed the expression of the potassium channel family Kv4, known to generate transient potassium currents upon depolarization. Such potassium currents are found in auditory efferent neurons, but it is not known whether Kv4 subunits are expressed in these neurons. Moreover, it is not known if Kv4 is present and has a function in the vestibular efferent neurons. Double labelling with anti-ChAT and anti-Kv4.2 or Kv4.3 demonstrates that the Kv4.3 subunits are abundantly expressed in audio-vestibular efferents, thus indicating that this subunit is a large contributor to the excitability and firing properties of the auditory efferent neurons, and most probably also for the vestibular efferent neurons. In addition, we also unexpectedly found a strong expression of Kv4.3 in principal cells of the superior olive, the neurons which are important for sound localization.</p>

Made available in DSpace on 2014-12-17T15:36:53Z (GMT). No. of bitstreams: 1 FrancimarAS.pdf: 4205737 bytes, checksum: a4c2113c8c56b274ed7600ae2a39f704 (MD5) Previous issue date: 2008-09-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The auditory system is composed by a set of relays from the outer ear to the cerebral cortex. In mammals, the central auditory system is composed by cochlear nuclei, superior olivary complex, inferior colliculus and medial geniculate body. In this study, the auditory rombencephalic centers, the cochlear nuclear complex and the superior olivary complex were evaluated from the cytoarchitecture and neurochemical aspects, thorough Nissl staining and immunohistochemical techniques to reveal specific neuron nuclear protein (NeuN), glutamate (Glu), glutamic acid decarboxilase (GAD), enkephalin (ENK), serotonin (5-HT), choline acetyltransferase (ChAT) and calcium-binding proteins calbindin (CB), calretinin (CR), and parvalbumin (PV). The common marmoset (Callithrix jacchus), a little native primate of the Brazilian atlantic forest was used as an experimental animal. As results, it was noted that the cochlear nuclear complex is composed by anteroventral, posteroventral and dorsal nuclei, and the superior olivary complex is constituted by the lateral and medial superior olivary nuclei and the trapezoid body nucleus. Glu, GAD, ENK, ChAT, CB, CR, PV-immunoreactive cells, fibers and terminals besides besides only 5-HT terminals were found unhomogeneously in all nuclei, of both complex. The emerging data are discussed in a comparative and functional context, and represent an important contribution to knowledge of the central auditory pathways in the common marmoset, and then in primates / O sistema auditivo compreende uma s?rie de esta??es que se estendem desde a orelha externa at? o c?rtex cerebral. Em mam?feros o sistema auditivo central subcortical ? formado essencialmente por n?cleos cocleares, complexo olivar superior, col?culo inferior e corpo geniculado medial. Neste estudo, os centros rombencef?licos, compreendendo o complexo nuclear coclear e o complexo olivar superior foram avaliados com rela??o a sua citoarquitetura e conte?do neuroqu?mico de corpos celulares e terminais ax?nicos, atrav?s das t?cnicas de colora??o de Nissl e imuno-histoqu?mica para prote?na nuclear neur?nio espec?fica (NeuN), glutamato (Glu), descaboxilase de ?cido glut?mico (GAD), encefalina (ENK), serotonina (5-HT), colina acetiltransferase (ChAT) e prote?nas ligantes de c?lcio calbindina (CB), cal-retinina (CR) e parvalbumina (PV). Foi utilizado como animal experimental o sag?i (Callithrix jacchus), um pequeno primata nativo da Mata Atl?ntica do Nordeste Brasileiro. Como resultados, foi evidenciado que o complexo nuclear coclear ? composto pelos n?cleos cocleares antero-ventral, p?stero-ventral e dorsal, e o complexo olivar superior pelos n?cleos olivares superiores lateral e medial e o n?cleo do corpo trapez?ide. Em todos os n?cleos, de ambos os complexos, foram encontrados de forma vari?vel corpos celulares, fibras e terminais imunorreativos a Glu, GAD, ChAT, CB, CR, PV, corpos celulares e terminais imunoreativos a ENK, al?m de fibras e terminais imunorreativos a 5-HT em diferentes densidades. Os dados obtidos s?o discutidos dentro de um contexto comparativo e funcional e representam uma importante contribui??o ao conhecimento das vias auditivas centrais no sag?i, e por extens?o em primatas

The neural code from the inner ear to the brain is dynamically controlled by central nervous efferent feedback to the audio-vestibular epithelium. Although such efference provides the basis for a cognitive control of our hearing and balance, we know surprisingly little about this feedback system. This project has investigated the applicability of a transgenic mouse model, expressing a fluorescent protein under the choline-acetyltransferase (ChAT) promoter, for targeting the cholinergic audio-vestibular efferent neurons in the brainstem. It was found that the mouse model is useful for targeting the vestibular efferents, which are fluorescent, but not the auditory efferents, which are not highlighted. This model enables, for the first time, physiological studies of the vestibular efferent neurons and their synaptic inputs. We next assessed the expression of the potassium channel family Kv4, known to generate transient potassium currents upon depolarization. Such potassium currents are found in auditory efferent neurons, but it is not known whether Kv4 subunits are expressed in these neurons. Moreover, it is not known if Kv4 is present and has a function in the vestibular efferent neurons. Double labelling with anti-ChAT and anti-Kv4.2 or Kv4.3 demonstrates that the Kv4.3 subunits are abundantly expressed in audio-vestibular efferents, thus indicating that this subunit is a large contributor to the excitability and firing properties of the auditory efferent neurons, and most probably also for the vestibular efferent neurons. In addition, we also unexpectedly found a strong expression of Kv4.3 in principal cells of the superior olive, the neurons which are important for sound localization.