Combined in situ hybridization chain reaction and immunostaining to visualize gene expression in whole-mount Drosophila central nervous systems

Methods to visualize gene expression in the Drosophila central nervous system are important in fly neurogenetic studies. In this chapter, we describe a detailed protocol that sequentially combines in situ hybridization chain reaction (HCR) and immunostaining to detect mRNA and protein expression in...

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Bibliographic Details
Published in:bioRxiv
Main Authors: Duckhorn, Julia C, Junker, Ian P, Ding, Yun, Shirangi, Troy R
Format: Paper
Language:English
Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 10.08.2021
Cold Spring Harbor Laboratory
Edition:1.1
Subjects:
Central nervous system
Drosophila
Gene expression
Hybridization
Insects
Kinases
Nervous system
Neuroscience
ISSN:2692-8205, 2692-8205
Online Access:Get full text
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Summary:Methods to visualize gene expression in the Drosophila central nervous system are important in fly neurogenetic studies. In this chapter, we describe a detailed protocol that sequentially combines in situ hybridization chain reaction (HCR) and immunostaining to detect mRNA and protein expression in whole-mount Drosophila larval and adult central nervous systems. We demonstrate the application of in situ HCR in comparisons of nervous system gene expression between Drosophila species, and in the validation of single-cell RNA-Seq results in the fly nervous system. Our protocol provides a simple, robust, multiplexable, and relatively affordable means to quantitatively visualize gene expression in the nervous system of flies, facilitating its general use in fly neurogenetic studies. Competing Interest Statement The authors have declared no competing interest.
Bibliography:SourceType-Working Papers-1
ObjectType-Working Paper/Pre-Print-1
content type line 50
Competing Interest Statement: The authors have declared no competing interest.
ISSN:2692-8205
2692-8205
DOI:10.1101/2021.08.02.454831