Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Expression profiles of globular bushy cells (GBCs) during maturation

ABSTRACT: Calyx of Held giant presynaptic terminals in the medial nucleus of the trapezoid body of the auditory brainstem form axosomatic synapses that have advanced to one of the best-studied synaptic system of the mammalian brain. As the auditory system matures and adjusts to high fidelity synaptic transmission, the calyx undergoes extensive structural and functional changes: it is formed around postnatal day 3 (P3), achieves immature function until hearing onset around P10 and can be considered mature from P21 onwards. This setting provides the unique opportunity to examine the repertoire of genes driving synaptic structure and function. We performed cell type-specific gene expression profiling of globular bushy cells (GBCs), the neurons giving rise to the calyx of Held, at different maturational stages (P3, P8 and P21). We identified GBCs by stereotaxic injection of fluorescently labelled retrograde tracer Cholera toxin B into the contralateral MNTB of anesthetized rats. Animals were sacrificed 24h after injection, the brain was taken out and flash frozen. 12um thick brainstem cryosections were prepared and 200 fluorescently labelled GBCs per animal were excised from the VCN using laser microdissection. Cells were collected from 6 animals at P3 (synapse formation), 9 animals at P8 (juvenile synapse) and 5 animals at P21 (mature synapse). RNA was isolated from the collected cells and linearly amplified in order to perform cell-type specific expression profiling.

ORGANISM(S): Rattus norvegicus

SUBMITTER: Christoph Koerber

PROVIDER: E-GEOD-49599 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Gene expression profile during functional maturation of a central mammalian synapse.

Körber Christoph C Dondzillo Anna A Eisenhardt Gisela G Herrmannsdörfer Frank F Wafzig Oliver O Kuner Thomas T

The European journal of neuroscience 20140703 6

Calyx of Held giant presynaptic terminals in the auditory brainstem form glutamatergic axosomatic synapses that have advanced to one of the best-studied synaptic connections of the mammalian brain. As the auditory system matures and adjusts to high-fidelity synaptic transmission, the calyx undergoes extensive structural and functional changes - in mice, it is formed at about postnatal day 3 (P3), achieves immature function until hearing onset at about P10 and can be considered mature from P21 on ...[more]

PMID: 24995587

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Project description:Spiral ganglion neurons (SGNs) and the associated components of the auditory nerve are primary carriers of auditory information from hair cells to the brain. Loss of SGNs occurs with many pathological conditions, resulting in permanent sensorineural hearing loss. Neural stem/progenitors (NSPs) have been well-characterized in several locations of adult brain and retina. However, it is unclear whether NSPs are present in the adult auditory nerve. Here we examined the self-renewal potential of the adult auditory nerve using ouabain application as a well-established mouse model of acute SGN injury. The observed increase in cell proliferation, alteration in enchromatin/heterochromatin ratio and down-regulation of histone deacetylase expression in glial cells suggest that the quiescent glial cells convert to an activated state after SGN degeneration. This was further confirmed by global gene expression analysis of injured auditory nerves, which showed up-regulation of numerous neurogenesis- and/or development-associated genes shortly after ouabain exposure. These genes include molecular markers commonly used for the identification of NSPs. Under a strict culture regimen, auditory nerve-derived cells of adult mouse ears gave rise to neurospheres, suggesting that multipotent NSPs are present in adult cochlear nerve. Neurosphere assays on Sox2 transgenic mice revealed that Sox2+ glial cells are the source for NSPs. Our data also showed that acute injury or hypoxia enhances neurosphere formation. Taken together, our study revealed that glial cells of adult cochlea exhibit several NSP characteristics, and hence these mature non-neuronal cells may be important targets for promoting self-repair of degenerative auditory nerves. Analysis was conducted on auditory nerve samples from stages encompassing maturation and onset of hearing. Cochleas were collected from euthanized mice (CBA/CaJ) at perinatal (P) stages P0, P3, P7, P10, P14 and P21. Cochleas underwent microdissection to remove the outer bony cochlear shell and cochlear lateral wall, thus preserving the modiolus portion of cochlea which contains mainly the auditory nerve. Paired cochleas (i.e., from left and right ears) from each mouse were pooled to make individual samples. All sample types were done in experimental duplicate (n=2).

Dataset Information

Expression profiles of globular bushy cells (GBCs) during maturation

Publications

Gene expression profile during functional maturation of a central mammalian synapse.

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