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

Mujeeb Shittu; Tessa Steenwinkel; William Dion; Nathan Ostlund; Komal Raja; Thomas Werner

doi:10.3390/mps4010020

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

Methods Protoc. 2021 Mar 10;4(1):20. doi: 10.3390/mps4010020.

Authors

Mujeeb Shittu¹, Tessa Steenwinkel¹, William Dion^{2

3}, Nathan Ostlund¹, Komal Raja⁴, Thomas Werner¹

Affiliations

¹ Department of Biological Sciences, Michigan Technological University, 740 Dow Building, Houghton, MI 49931, USA.
² Integrative Systems Biology Graduate Program, University of Pittsburgh School of Medicine, Biomedical Science, Tower 3, 3501 Fifth Avenue, Pittsburgh, PA 15213, USA.
³ Aging Institute of UPMC, University of Pittsburgh School of Medicine, Bridgeside Point 1, 5th Floor, 100 Technology Drive, Pittsburgh, PA 15219, USA.
⁴ Department of Pathology and Immunology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA.

Abstract

RNA in situ 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 Drosophila 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 Drosophila species.

Keywords: Drosophila; gene expression; in situ hybridization.

Grants and funding

1R15GM107801-01A1/NH/NIH HHS/United States