Detection of single mRNAs in individual cells of the auditory system.

Title

Detection of single mRNAs in individual cells of the auditory system.

Creator

Salehi Pezhman; Nelson Charlie N; Chen Yingying; Lei Debin; Crish Samuel D; Nelson Jovitha; Zuo Hongyan; Bao Jianxin

Publisher

Hearing research

Date

2018
2018-09

Description

Gene expression analysis is essential for understanding the rich repertoire of cellular functions. With the development of sensitive molecular tools such as single-cell RNA sequencing, extensive gene expression data can be obtained and analyzed from various tissues. Single-molecule fluorescence in situ hybridization (smFISH) has emerged as a powerful complementary tool for single-cell genomics studies because of its ability to map and quantify the spatial distributions of single mRNAs at the subcellular level in their native tissue. Here, we present a detailed method to study the copy numbers and spatial localizations of single mRNAs in the cochlea and inferior colliculus. First, we demonstrate that smFISH can be performed successfully in adult cochlear tissue after decalcification. Second, we show that the smFISH signals can be detected with high specificity. Third, we adapt an automated transcript analysis pipeline to quantify and identify single mRNAs in a cell-specific manner. Lastly, we show that our method can be used to study possible correlations between transcriptional and translational activities of single genes. Thus, we have developed a detailed smFISH protocol that can be used to study the expression of single mRNAs in specific cell types of the peripheral and central auditory systems.

Subject

Cochlea; Immunohistochemistry; Inner hair cell; Outer hair cell; Single-molecule fluorescence in situ hybridization; Spiral ganglion neuron

Rights

Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).

Pages

88–96

Volume

367

Citation

Salehi Pezhman; Nelson Charlie N; Chen Yingying; Lei Debin; Crish Samuel D; Nelson Jovitha; Zuo Hongyan; Bao Jianxin, “Detection of single mRNAs in individual cells of the auditory system.,” NEOMED Bibliography Database, accessed December 1, 2021, https://neomed.omeka.net/items/show/3587.

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