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用于解剖果蝇蛹并使用杂交链式反应可视化RNA表达的实验方案。

Protocol for dissecting Drosophila pupae and visualizing RNA expression using hybridization chain reaction.

作者信息

Bayala Erick X, Sinha Pratyush, Wittkopp Patricia J

机构信息

Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA.

Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103456. doi: 10.1016/j.xpro.2024.103456. Epub 2024 Nov 21.

DOI:10.1016/j.xpro.2024.103456
PMID:39576730
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11616507/
Abstract

Visualizing RNA expression in the Drosophila epidermis during pupal development is challenging because the tissue is fragile during early pupal stages and increasingly impermeable at later stages. Here, we present a protocol for tissue dissection and detection of RNA in situ. We describe steps for using the hybridization chain reaction (HCR) in early and late Drosophila pupal stages (and larval imaginal discs). This protocol facilitates the study of dynamically changing patterns in gene expression during pupal development.

摘要

在蛹发育过程中可视化果蝇表皮中的RNA表达具有挑战性，因为该组织在蛹早期阶段很脆弱，而在后期则越来越难以渗透。在这里，我们提出了一种组织解剖和RNA原位检测的方案。我们描述了在果蝇蛹早期和晚期（以及幼虫成虫盘）使用杂交链式反应（HCR）的步骤。该方案有助于研究蛹发育过程中基因表达的动态变化模式。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/580b/11616507/13fdb46126d2/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/580b/11616507/88cfbba51c4e/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/580b/11616507/9baa897de767/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/580b/11616507/13fdb46126d2/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/580b/11616507/88cfbba51c4e/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/580b/11616507/9baa897de767/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/580b/11616507/13fdb46126d2/gr2.jpg

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Development. 2024 Feb 15;151(4). doi: 10.1242/dev.202307. Epub 2024 Feb 28.
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Molecular evidence of anteroposterior patterning in adult echinoderms.分子证据表明成年棘皮动物具有前后模式。
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Dev Biol. 2023 Jun;498:87-96. doi: 10.1016/j.ydbio.2023.03.008. Epub 2023 Mar 24.
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A timer gene network is spatially regulated by the terminal system in the embryo.一个定时器基因网络通过胚胎中的终端系统在空间上被调控。
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