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Cell Biology JoVE (Open Access) Citable · DOI

An Injury Paradigm to Investigate Central Nervous System Repair in Drosophila

DOI: 10.3791/50306-v
What you'll learn
  • ✓ Perform precise stab injury to Drosophila larval ventral nerve cord
  • ✓ Acquire time-lapse confocal images of CNS regeneration in vivo
  • ✓ Quantify molecular markers of axonal repair using custom software
Protocol

An injury paradigm using the Drosophila larval ventral nerve cord to investigate central nervous system regeneration and repair is described. Stabbing followed by laser scanning confocal microscopy in time-lapse and fixed specimens, combined with quantitative analysis with purposefully developed software and genetics, are used to investigate the molecular mechanisms of CNS regeneration and repair.

Difficulty
advanced
Total time
~3–4 hours per larva (dissection through imaging and analysis)
Model organism
Drosophila melanogaster larva
Biosafety
BSL-1

Steps

1
Dissect larval ventral nerve cord tissue

Extract and expose the Drosophila larval VNC using standard dissection techniques to prepare the tissue for injury.

▶ 01:18
2
Deliver controlled stab injury to VNC

Use a fine needle or micro-manipulator to create a standardized stab wound in the ventral nerve cord, establishing the injury paradigm for regeneration studies.

▶ 03:56
3
Perform time-lapse confocal microscopy imaging

Acquire sequential confocal laser scanning images of the injured VNC over time to visualize dynamic regeneration and axonal repair processes.

▶ 06:07
4
Analyze post-injury ventral nerve cord images

Quantify regeneration markers and structural changes in fixed and live specimens using custom-developed software and genetic tools to assess repair mechanisms.

▶ 08:05
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