Profiling mouse cochlear cell maturation using 10× Genomics single-cell transcriptomics.

Profiling mouse cochlear cell maturation using 10× Genomics single-cell transcriptomics.
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DOI:
10.3389/fncel.2022.962106
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发表时间:
2022
影响因子:
5.3
通讯作者:
--
中科院分区:
医学2区
文献类型:
--
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幼年和成熟的小鼠耳蜗含有各种低丰度的,脆弱的感觉上皮细胞嵌入钙化的颞骨,使其具有挑战性的轮廓在单细胞水平上的成熟过程中,这些细胞的动态转录组变化。在这里,我们在出生后第14天(P14)和第28天对小鼠耳蜗进行了10 x Genomics单细胞RNA测序(scRNA-seq)。我们获得了多种细胞类型的转录组,包括毛细胞、支持细胞、螺旋神经节、纤维纹细胞和免疫细胞。我们的毛细胞scRNA-seq数据集与已发表的来自批量RNA-seq的转录物一致。我们还将已知的耳聋基因映射到相应的耳蜗细胞类型。重要的是,伪时间轨迹分析显示,内毛细胞成熟在P14达到峰值,而外毛细胞继续发育,直到P28。我们进一步鉴定并证实了一个长的非编码RNA基因Miat在耳蜗毛细胞和螺旋神经节神经元的成熟过程中表达,而Pcp 4在耳蜗毛细胞的成熟过程中表达。我们的少年和成熟的小鼠耳蜗细胞的转录组提供的续集,以前发表的胚胎晚期和出生后早期的年龄,将是宝贵的资源,以调查耳蜗成熟的单细胞分辨率。
Juvenile and mature mouse cochleae contain various low-abundant, vulnerable sensory epithelial cells embedded in the calcified temporal bone, making it challenging to profile the dynamic transcriptome changes of these cells during maturation at the single-cell level. Here we performed the 10x Genomics single-cell RNA sequencing (scRNA-seq) of mouse cochleae at postnatal days 14 (P14) and 28. We attained the transcriptomes of multiple cell types, including hair cells, supporting cells, spiral ganglia, stria fibrocytes, and immune cells. Our hair cell scRNA-seq datasets are consistent with published transcripts from bulk RNA-seq. We also mapped known deafness genes to corresponding cochlear cell types. Importantly, pseudotime trajectory analysis revealed that inner hair cell maturation peaks at P14 while outer hair cells continue development until P28. We further identified and confirmed a long non-coding RNA gene Miat to be expressed during maturation in cochlear hair cells and spiral ganglia neurons, and Pcp4 to be expressed during maturation in cochlear hair cells. Our transcriptomes of juvenile and mature mouse cochlear cells provide the sequel to those previously published at late embryonic and early postnatal ages and will be valuable resources to investigate cochlear maturation at the single-cell resolution.
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