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成年小鼠血管纹的解剖用于单细胞测序或免疫染色。

Dissection of Adult Mouse Stria Vascularis for Single-Nucleus Sequencing or Immunostaining.

机构信息

Auditory Development and Restoration Program, National Institute on Deafness and Other Communication Disorders, National Institutes of Health.

Auditory Development and Restoration Program, National Institute on Deafness and Other Communication Disorders, National Institutes of Health;

出版信息

J Vis Exp. 2023 Apr 21(194). doi: 10.3791/65254.

DOI:10.3791/65254
PMID:37154552
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10443831/
Abstract

Endocochlear potential, which is generated by the stria vascularis, is essential to maintain an environment conducive to appropriate hair cell mechanotransduction and ultimately hearing. Pathologies of the stria vascularis can result in a decreased hearing. Dissection of the adult stria vascularis allows for focused single-nucleus capture and subsequent single-nucleus sequencing and immunostaining. These techniques are used to study stria vascularis pathophysiology at the single-cell level. Single-nucleus sequencing can be used in the setting of transcriptional analysis of the stria vascularis. Meanwhile, immunostaining continues to be useful in identifying specific populations of cells. Both methods require proper stria vascularis dissection as a prerequisite, which can prove to be technically challenging.

摘要

内淋巴电位由血管纹产生,对于维持有利于毛细胞机械转导的环境至关重要,而毛细胞机械转导最终又与听觉相关。血管纹的病变可导致听力下降。成年血管纹的解剖可以实现单个核细胞的聚焦捕获,进而进行单个核细胞测序和免疫染色。这些技术可用于在单细胞水平上研究血管纹的病理生理学。单个核细胞测序可用于血管纹的转录分析。同时,免疫染色在鉴定特定细胞群方面仍然有用。这两种方法都需要适当的血管纹解剖作为前提,这可能具有一定的技术挑战性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebb3/10443831/9e2fa8be00e5/nihms-1921971-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebb3/10443831/2c489657f892/nihms-1921971-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebb3/10443831/d4e2ba00a7cf/nihms-1921971-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebb3/10443831/91c4afd9ce59/nihms-1921971-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebb3/10443831/9e2fa8be00e5/nihms-1921971-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebb3/10443831/2c489657f892/nihms-1921971-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebb3/10443831/d4e2ba00a7cf/nihms-1921971-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebb3/10443831/91c4afd9ce59/nihms-1921971-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebb3/10443831/9e2fa8be00e5/nihms-1921971-f0004.jpg

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本文引用的文献

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Perspectives on single-nucleus RNA sequencing in different cell types and tissues.不同细胞类型和组织中单核RNA测序的研究视角。
J Pathol Transl Med. 2023 Jan;57(1):52-59. doi: 10.4132/jptm.2022.12.19. Epub 2023 Jan 10.
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The role of the stria vascularis in neglected otologic disease.血管纹在被忽视的耳科学疾病中的作用。
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In vivo real-time imaging reveals megalin as the aminoglycoside gentamicin transporter into cochlea whose inhibition is otoprotective.
体内实时成像显示 megalin 是氨基糖苷类抗生素庆大霉素进入耳蜗的转运体,其抑制具有耳保护作用。
Proc Natl Acad Sci U S A. 2022 Mar 1;119(9). doi: 10.1073/pnas.2117946119.
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Tissue-Resident Macrophages in the Stria Vascularis.血管纹中的组织驻留巨噬细胞
Front Neurol. 2022 Feb 3;13:818395. doi: 10.3389/fneur.2022.818395. eCollection 2022.
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Single nucleus RNA-sequencing: how it's done, applications and limitations.单细胞 RNA 测序:技术原理、应用及局限性
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Nat Rev Genet. 2021 Oct;22(10):627-644. doi: 10.1038/s41576-021-00370-8. Epub 2021 Jun 18.
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VEGFA165 gene therapy ameliorates blood-labyrinth barrier breakdown and hearing loss.VEGFA165 基因治疗可改善血迷路屏障破坏和听力损失。
JCI Insight. 2021 Apr 22;6(8):143285. doi: 10.1172/jci.insight.143285.
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