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Updated: Jul 14, 2025

5-Ethynyl-2'-Deoxyuridine/Phospho-Histone H3 Dual-Labeling Protocol for Cell Cycle Progression Analysis in Drosophila Neural Stem Cells

5-Ethynyl-2'-Deoxyuridine/Phospho-Histone H3 Dual-Labeling Protocol for Cell Cycle Progression Analysis in Drosophila Neural Stem Cells

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A modified method to analyse cell proliferation using EdU labelling in large insect brains.

Amaia Alcalde Anton¹, Max S Farnworth¹, Laura Hebberecht¹

¹School of Biological Sciences, University of Bristol, Bristol, United Kingdom.

|October 5, 2023

Summary

Neurogenesis, the creation of new neurons, is key to understanding nervous system evolution. This study adapts EdU staining for studying neurogenesis in large-brained insects like Heliconiini butterflies.

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5-Ethynyl-2'-Deoxyuridine/Phospho-Histone H3 Dual-Labeling Protocol for Cell Cycle Progression Analysis in Drosophila Neural Stem Cells

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Cell Cycle Analysis in the C. elegans Germline with the Thymidine Analog EdU

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Area of Science:

Evolutionary biology
Neuroscience
Developmental biology

Background:

Neurogenesis is crucial for understanding nervous system evolution.
Drosophila melanogaster is a common model, but insect nervous systems show high diversity.
Mushroom bodies, vital for learning and memory, vary significantly across insect species.

Purpose of the Study:

To investigate the evolution of nervous systems by studying neurogenesis in diverse insect taxa.
To adapt and present a modified EdU staining protocol for examining neurogenesis in large-brained insects.
To explore the variation in mushroom body size and its relation to Kenyon cell neurogenesis in Heliconiini butterflies.

Main Methods:

Modified EdU staining protocol.
Application of the protocol to Heliconiini butterflies and cockroaches.
Analysis of neurogenesis in Kenyon cells, the intrinsic neurons of mushroom bodies.

Main Results:

The study successfully adapted EdU staining for large-brained insects.
Demonstrated the protocol's applicability in Heliconiini butterflies and cockroaches.
Provided insights into the variation of mushroom body size linked to Kenyon cell proliferation.

Conclusions:

The adapted EdU staining protocol is effective for studying neurogenesis in understudied, large-brained insects.
This method facilitates research into the evolution of diverse nervous systems.
Understanding Kenyon cell neurogenesis in Heliconiini butterflies offers insights into brain evolution.