RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> Species

RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> Species

RNA <i>in situ</i> hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model <i>Drosophila</i> sp...

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Main Authors: Mujeeb Shittu, Tessa Steenwinkel, William Dion, Nathan Ostlund, Komal Raja, Thomas Werner
Format: Article
Language:English
Published: MDPI AG 2021-03-01
Series:Methods and Protocols
Subjects:
<i>Drosophila</i>
<i>in situ</i> hybridization
gene expression
Online Access:https://www.mdpi.com/2409-9279/4/1/20
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spelling doaj-245d4563b10544e0b0684b151e0bfacb2021-03-11T00:00:21ZengMDPI AGMethods and Protocols2409-92792021-03-014202010.3390/mps4010020RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> SpeciesMujeeb Shittu0Tessa Steenwinkel1William Dion2Nathan Ostlund3Komal Raja4Thomas Werner5Department of Biological Sciences, Michigan Technological University, 740 Dow Building, Houghton, MI 49931, USADepartment of Biological Sciences, Michigan Technological University, 740 Dow Building, Houghton, MI 49931, USAIntegrative Systems Biology Graduate Program, University of Pittsburgh School of Medicine, Biomedical Science, Tower 3, 3501 Fifth Avenue, Pittsburgh, PA 15213, USADepartment of Biological Sciences, Michigan Technological University, 740 Dow Building, Houghton, MI 49931, USADepartment of Pathology and Immunology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USADepartment of Biological Sciences, Michigan Technological University, 740 Dow Building, Houghton, MI 49931, USARNA <i>in situ</i> hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model <i>Drosophila</i> species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model <i>Drosophila</i> species.https://www.mdpi.com/2409-9279/4/1/20<i>Drosophila</i><i>in situ</i> hybridizationgene expression
collection DOAJ
language English
format Article
sources DOAJ
author Mujeeb Shittu
Tessa Steenwinkel
William Dion
Nathan Ostlund
Komal Raja
Thomas Werner
spellingShingle Mujeeb Shittu
Tessa Steenwinkel
William Dion
Nathan Ostlund
Komal Raja
Thomas Werner
RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> Species
Methods and Protocols
<i>Drosophila</i>
<i>in situ</i> hybridization
gene expression
author_facet Mujeeb Shittu
Tessa Steenwinkel
William Dion
Nathan Ostlund
Komal Raja
Thomas Werner
author_sort Mujeeb Shittu
title RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> Species
title_short RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> Species
title_full RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> Species
title_fullStr RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> Species
title_full_unstemmed RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> Species
title_sort rna <i>in situ</i> hybridization for detecting gene expression patterns in the abdomens and wings of <i>drosophila</i> species
publisher MDPI AG
series Methods and Protocols
issn 2409-9279
publishDate 2021-03-01
description RNA <i>in situ</i> hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model <i>Drosophila</i> species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model <i>Drosophila</i> species.
topic <i>Drosophila</i>
<i>in situ</i> hybridization
gene expression
url https://www.mdpi.com/2409-9279/4/1/20
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