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RiboTag translatome analysis of outer hair cell postnatal development

ABSTRACT: RiboTag translatome analysis of outer hair cell postnatal development

PROVIDER: PRJNA479956 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7477280_1.fastq.gz	Fastqsanger.gz
	SRR7477280_2.fastq.gz	Fastqsanger.gz
	SRR7477290_1.fastq.gz	Fastqsanger.gz
	SRR7477290_2.fastq.gz	Fastqsanger.gz
	SRR7477300_1.fastq.gz	Fastqsanger.gz

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RiboTag translatome analysis of outer hair cell postnatal development

Project description:In order to determine the regulators of outer hair cell postnatal maturation, we utilized the RiboTag mouse model to perform a detailed transcriptomic analysis of outer hair cells at five postnatal developmental time points: P8, P14, P28, 6 weeks (6wk) and 10 weeks (10wk). This analysis resulted in consistent enrichment of outer hair cell expressed genes in the immunoprecipitated RNA compared to whole cochlear input RNA from each time point. Using transcription factor binding motif prediction on a set of defined outer hair cell enriched genes, we further use this dataset to identify the helios transcription factor as a regulator of the postnatal outer hair cell transcriptome.

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Project description:Given the potential role of the helios transcription factor in postnatal outer hair cell gene regulation indicated by our RiboTag translatome analyses, our next goal was to identify and validate genes that are regulated by helios in outer hair cells by performing RNA-seq on P8 cello mutant animals compared to wild-type littermates.

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Project description:To further understand the biological properties of hair cells of the mammalian cochlea, we examined the transcriptome of adult inner and outer hair cells. Morphologically distinct inner and outer hair cells were isolated from the organ of Corti from adult CBA/J mice. One thousand inner and outer hair cells were separately collected for each biological replicate, using the suction pipette technique. RNA sequencing of two biological replicates of IHCs and three biological replicates of OHCs, each with two technical repeats, was performed. The resulting sequenced reads were mapped. Comparisons between inner and outer hair cells allow identification of enriched genes, as well as differentially expressed genes that result in cellular specialization. Our dataset provides an extensive resource for understanding the molecular mechanisms underlying morphology, function, and pathology of adult mouse inner and outer hair cells.

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Project description:In order to identify the transcriptional cascade downstream of the GFI1 repressor in inner ear hair cells, we performed a RiboTag-based expression analysis of Gfi1cre/cre mutant hair cells compared to their heterozygous controls. Through this analysis, we identified a significant downregulation of genes involved in normal hair cell development and function within the mutant hair cells, suggesting that in the absence of GFI1, the hair cells undergo a maturation arrest. Furthermore, the Gfi1 deficient hair cells displayed a significant upregulation of genes involved in neurodifferentiation and neuronal function.

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