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Título: Validation of optogenetic protein expression in the Dorsal cochlear nucleus: molecular basis for in vitro and in vivo investigation of tinnitus in mice
Título(s) alternativo(s): Validação da expressão de proteínas optogenéticas no Núcleo coclear dorsal: bases moleculares para investigação in vitro e in vivo de tinnitus em camundongos
Autor(es): Borges, Thawann Malfatti
Palavras-chave: Optogenetics;Dorsal cochlear nucleus;Auditory system;Tinnitus
Data do documento: 26-Jun-2015
Editor: Universidade Federal do Rio Grande do Norte
Citação: BORGES, Thawann Malfatti. Validation of optogenetic protein expression in the Dorsal cochlear nucleus: molecular basis for in vitro and in vivo investigation of tinnitus in mice. 2015. 60f. Dissertação (Mestrado em Neurociências) - Universidade Federal do Rio Grande do Norte, Natal, 2015.
metadata.dc.description.resumo: Tinnitus is the perception of a sound in the absence of a corresponding physical stimulus. It is not clear yet what mechanisms are involved in tinnitus and how it starts and/or becomes chronic. Due to the relationship between tinnitus and somatosensory trauma/stimuli, the dorsal cochlear nucleus (DCN), a region known to integrate somatosensory and auditory pathways, has been identified as a potential key structure in the generation of phantom sound perception. Here, we target specific neuronal populations in the DCN to allow further investigation on how this region may contribute to the generation of tinnitus signals that spread to other auditory areas. We examined the expression of optogenetic proteins (Channelrhodopsin 2 - ChR2; and enhanced Archaerhodopsin 3.0 - eArch3.0), targeting neurons expressing Calmoduline Kinase II alpha (CaMKIIa) promoter in wild-type C57/Bl6 mice and neurons expressing nicotinic acetylcholine receptor subunit alpha-2 promoter (ChRNA2) in ChRNA2- Cre transgenic C57/Bl6 mice, using local virus injection, verified by fluorescence microscopy. Unit responses were differentiated based on their electrophysiological response to sound. We then investigated if firing of neurons expressing optogenetic tools can be controlled in vivo and if the same neurons also fire action potentials in response to precisely timed sound stimulation. Both in vivo and preliminary in vitro data shows that neurons expressing ChR2 do respond to sound, and that they furthermore also can respond to light stimulation with a stable and similar waveform. Moreover, in vivo data shows that neurons expressing eArch3.0, responding to sound, will decrease their firing rate when exposed to green light. Thereby showing that optogenetic tools can be used functionally in the DCN, it is possible to further test tinnitus theories by, for example, producing an increased firing rate in the DCN, trying to mimic tinnitus; or inhibiting increased spontaneous firing rate in the DCN of animals with noise-induced or salycilate-induced tinnitus.
Aparece nas coleções:PPGNEURO - Mestrado em Neurociências

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