Insights into the Biology of Hearing and Deafness Revealed by Single-Cell RNA Sequencing
Insights into the Biology of Hearing and Deafness Revealed by Single-Cell RNA Sequencing
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DOI：
10.1016/j.celrep.2019.02.053
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发表时间：
2019-03-12
期刊：
影响因子：
8.8
通讯作者：
Smith, Richard J. H.
中科院分区：
文献类型：
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作者：
Ranum, Paul T.;Goodwin, Alexander T.;Smith, Richard J. H.
单细胞 RNA 测序是一种强大的工具，可用于表征低丰度细胞类型的转录谱，但其在内耳的应用受到骨迷路、组织稀疏以及难以分离膜性耳蜗极其稀有细胞的阻碍。在此，我们提出了一种在出生后任何时间点从小鼠耳蜗中分离单个内毛细胞 (IHC)、外毛细胞 (OHC) 和 Deiters 细胞 (DC) 的方法。我们从出生后第 15 天 (p15) 到第 228 天 (p228) 收获了 200 多个小鼠 IHC、OHC 和 DC，并利用短读长和长读单细胞 RNA 测序来分析转录物丰度和结构。我们的结果提供了对这些细胞表达谱的深入了解，并记录了耳聋相关基因异构体多样性未被认识到的复杂性。这种对柯蒂氏器转录的精细观察提高了我们对听力和耳聋生物学的理解。
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 ultrarare 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.