Immunohistochemistry and Fluorescent Whole Mount RNA In Situ Hybridization in Larval and Adult Brains of Tribolium

  • Vera S. Hunnekuhl
  • Janna Siemanowski
  • Max S. Farnworth
  • Bicheng He
  • Gregor BucherEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2047)


Arthropod brains are fascinating structures that exhibit great complexity but also contain conserved elements that can be recognized between species. There is a long tradition of research in insect neuroanatomy, cell biology, and in studying the genetics of insect brain development. Recently, the beetle Tribolium castaneum has gained attention as a model for insect head and brain development, and many anterior patterning genes have so far been characterized in beetle embryos. The outcome of embryonic anterior development is the larval and, subsequently, the adult brain. A basic requirement to understand genetic cell type diversity within these structures is the ability to localize mRNA and protein of neural genes. Here we detail our protocols for RNA in situ hybridization in combination with immunohistochemistry, optimized for dissected brains of larval and adult beetles.


In situ hybridization Gene expression Antibody staining Fluorescence labeling Insect brain Tribolium castaneum Red flour beetle 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Vera S. Hunnekuhl
    • 1
  • Janna Siemanowski
    • 1
  • Max S. Farnworth
    • 1
  • Bicheng He
    • 1
  • Gregor Bucher
    • 1
    Email author
  1. 1.Department of Evolutionary Developmental GeneticsGeorg-August-University GöttingenGöttingenGermany

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