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Título: Using cortical neuron markers to target cells in the dorsal cochlear nucleus
Autor(es): Borges, Thawann Malfatti
Boerner, Barbara Ciralli
Hilscher, Markus M
Edwards, Steven J.
Kullander, Klas
Leão, Richardson Naves
Leão, Emelie Katarina Svahn
Palavras-chave: CaMKIIa;Chrna2;Dorsal cochlear nucleus;Extracellular recording;Optogenetics;Ventral cochlear nucleus
Data do documento: 9-Fev-2021
Editor: Society for Neuroscience
Referência: MALFATTI, Thawann; CIRALLI, Barbara; HILSCHER, Markus M.; EDWARDS, Steven J.; KULLANDER, Klas; LEAO, Richardson N.; LEAO, Katarina E. Using cortical neuron markers to target cells in the dorsal cochlear nucleus. eNeuro, [S. l.], p. ENEURO.0413-20.2020, fev. 2021. Doi: Disponível em: Acesso em: 19 fev. 2021.
Resumo: The dorsal cochlear nucleus (DCN) is a region of particular interest for auditory and tinnitus research. Yet, lack of useful genetic markers for in vivo manipulations hinders elucidation of the DCN contribution to tinnitus pathophysiology. This work assesses whether adeno-associated viral vectors (AAV) containing the calcium/calmodulin-dependent protein kinase 2 alpha (CaMKIIα) promoter and a mouse line of nicotinic acetylcholine receptor alpha 2 subunit (Chrna2)-Cre can target specific DCN populations. We found that CaMKIIα cannot be used to target excitatory fusiform DCN neurons as labelled cells showed diverse morphology indicating they belong to different classes of DCN neurons. Light stimulation after driving Channelrhodopsin2 by the CaMKIIα promoter generated spikes in some units but firing rate decreased when light stimulation coincided with sound. Expression and activation of CaMKIIα-eArchaerhodopsin3.0 in the DCN produced inhibition in some units but sound-driven spikes were delayed by concomitant light stimulation. We explored the existence of Cre+ cells in the DCN of Chrna2-Cre mice by hydrogel embedding technique (CLARITY). There were almost no Cre+ cell bodies in the DCN; however, we identified profuse projections arising from the ventral cochlear nucleus (VCN). Anterograde labeling in the VCN revealed projections to the ipsilateral superior olive and contralateral medial nucleus of the trapezoid body (bushy cells); and a second bundle terminating in the DCN, suggesting the latter to be excitatory Chrna2+ T-stellate cells. Exciting Chrna2+ cells increased DCN firing. This work shows that cortical molecular tools may be useful for manipulating the DCN especially for tinnitus studies
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