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Text
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URL Address
<a href="http://doi.org/10.1016/j.heares.2018.07.008" target="_blank" rel="noreferrer noopener">http://doi.org/10.1016/j.heares.2018.07.008</a>
Pages
88–96
Volume
367
Dublin Core
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Title
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Detection of single mRNAs in individual cells of the auditory system.
Publisher
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Hearing research
Date
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2018
2018-09
Subject
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Cochlea; Immunohistochemistry; Inner hair cell; Outer hair cell; Single-molecule fluorescence in situ hybridization; Spiral ganglion neuron
Creator
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Salehi Pezhman; Nelson Charlie N; Chen Yingying; Lei Debin; Crish Samuel D; Nelson Jovitha; Zuo Hongyan; Bao Jianxin
Description
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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.
Identifier
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<a href="http://doi.org/10.1016/j.heares.2018.07.008" target="_blank" rel="noreferrer noopener">10.1016/j.heares.2018.07.008</a>
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Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
2018
Bao Jianxin
Chen Yingying
Cochlea
Crish Samuel D
Department of Anatomy & Neurobiology
Department of Pharmaceutical Sciences
Hearing research
Immunohistochemistry
Inner hair cell
Lei Debin
Nelson Charlie N
Nelson Jovitha
NEOMED College of Medicine
NEOMED College of Pharmacy
Outer hair cell
Salehi Pezhman
Single-molecule fluorescence in situ hybridization
Spiral ganglion neuron
Zuo Hongyan