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Cell Rep. 2019 Mar 12;26(11):3160-3171.e3. doi: 10.1016/j.celrep.2019.02.053.

Insights into the Biology of Hearing and Deafness Revealed by Single-Cell RNA Sequencing.

Author information

1
Interdisciplinary Graduate Program in Molecular & Cellular Biology, University of Iowa Graduate College, University of Iowa, Iowa City, IA 52242, USA; Molecular Otolaryngology and Renal Research Laboratories, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
2
Molecular Otolaryngology and Renal Research Laboratories, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
3
Molecular Otolaryngology and Renal Research Laboratories, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA; Department of Otorhinolaryngology, Shinshu University School of Medicine, Matsumoto, Japan.
4
Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, NE 68178, USA.
5
Interdisciplinary Graduate Program in Molecular & Cellular Biology, University of Iowa Graduate College, University of Iowa, Iowa City, IA 52242, USA; Molecular Otolaryngology and Renal Research Laboratories, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA; Department of Otolaryngology, Head and Neck Surgery, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA. Electronic address: richard-smith@uiowa.edu.

Abstract

Single-cell RNA sequencing is a powerful tool by which to characterize the transcriptional profile of low-abundance cell types, but its application to the inner ear has been hampered by the bony labyrinth, tissue sparsity, and difficulty dissociating the ultra-rare cells of the membranous cochlea. Herein, we present a method to isolate individual inner hair cells (IHCs), outer hair cells (OHCs), and Deiters' cells (DCs) from the murine cochlea at any post-natal time point. We harvested more than 200 murine IHCs, OHCs, and DCs from post-natal days 15 (p15) to 228 (p228) and leveraged both short- and long-read single-cell RNA sequencing to profile transcript abundance and structure. Our results provide insights into the expression profiles of these cells and document an unappreciated complexity in isoform variety in deafness-associated genes. This refined view of transcription in the organ of Corti improves our understanding of the biology of hearing and deafness.

KEYWORDS:

RNA sequencing; RNA-seq; alternative splicing; auditory; cochlea; gene expression; hair cells; hearing; isoforms; scRNA-seq; single-cell RNA-seq; sorcin; splicing

PMID:
30865901
DOI:
10.1016/j.celrep.2019.02.053
Free PMC Article

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