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Coupling clearing and Hybridization Chain Reaction approaches to investigate gene expression in organs inside whole-mount intact insect.
  • Bastien Cayrol,
  • Stefano Colella,
  • Marilyne Uzest
Bastien Cayrol
Univ Montpellier
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Stefano Colella
Univ Montpellier
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Marilyne Uzest
Univ Montpellier

Corresponding Author:[email protected]

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Detecting RNA molecules within their natural environment inside intact arthropods has long been challenging, particularly in small organisms covered by a tanned and pigmented cuticle. Here, we have developed a methodology that enables high-resolution analysis of the spatial distribution of transcripts of interest without having to dissect tiny organs or tissues, thereby preserving their integrity. We have combined an in situ amplification approach based on Hybridization Chain Reaction, which enhances the signal-to-noise ratio, and a clearing approach that allows the visualization of inner organs beneath the cuticle. We have implemented this methodology for the first time in Hemiptera, mapping two salivary aphid ( Acyrthosiphon pisum) transcripts, the effector c002 and the salivary sheath protein SHP. With a multiplex approach, we could simultaneously detect different mRNAs in whole-mount pea aphid head-thorax samples and show that they were distributed in distinct secretory cells of salivary glands.
Submitted to Microscopy Research and Technique
29 Feb 20241st Revision Received
04 Mar 2024Submission Checks Completed
04 Mar 2024Assigned to Editor
04 Mar 2024Review(s) Completed, Editorial Evaluation Pending
14 Mar 2024Editorial Decision: Accept
Apr 2024Published in Microscopy Research and Technique. 10.1002/jemt.24561