Endogenous patterned activity-regulated genes specifically regulate different phases of synaptogenesis in the auditory system
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ABSTRACT:
PROVIDER: PRJNA1195347 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR31639800_1.fastq.gz | Fastqsanger.gz | |||
| SRR31639800_2.fastq.gz | Fastqsanger.gz | |||
| SRR31639801_1.fastq.gz | Fastqsanger.gz | |||
| SRR31639801_2.fastq.gz | Fastqsanger.gz | |||
| SRR31639802_1.fastq.gz | Fastqsanger.gz |
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Project description:In the auditory system, spatiotemporal patterning of correlated activity that originates in immature hair cells (HC) instructs refinement of synaptic connections and tonotopy. Patterned activity-dependent synaptic pruning requires the interplay between Hebbian plasticity and activity-regulated genes (ARGs). However, the genetic basis of this process is incompletely understood. Here we demonstrate that developing spiral ganglion neurons (SGNs) transiently generate discrete bursts of spikes and that blocking burst activity in this critical window causes profound deafness in mice. Using RNA sequencing, we profile ARG programs by genetically disrupting endogenous activity patterns of SGNs without changing spontaneous firing of inner HCs. Gene knockdown reveals that Nptx2 mediates initial synaptic formations at early stage, whereas Them5, which is important for cardiolipin remodeling that enhances mitochondrial function, specifically regulates synaptic pruning at later stage of development. These results suggest that ARGs dictate different phases of synaptogenesis and that Them5-depedent mitochondrial function might underlie synaptic pruning.
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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. Auditory nerves were removed from the temporal bones of adult CBA/CaJ mice, aged 8 to 12 weeks. Tissues were either collected and used directly as the tissue samples or dissociated and used for the cell culture samples. Dissociated auditory nerve cells were propagated and grown to full confluency (5-7 days), constituting the cultured cell samples. For neurosphere samples, growth medium was changed to neurosphere formation medium and the cells were cultured for an additional 12 days. All samples were prepared in triplicate (n=3).
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