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Microbiology JoVE (Open Access) Citable · DOI

An Anoxia-starvation Model for Ischemia/Reperfusion in C. elegans

DOI: 10.3791/51231-v
What you'll learn
  • ✓ Construct a lab-made hypoxic chamber for anoxia induction in C. elegans.
  • ✓ Perform anoxia/starvation treatment and assess worm viability and touch responses.
  • ✓ Quantify neuronal damage via GFP imaging and behavioral functional assays.
  • ✓ Model ischemia/reperfusion injury using C. elegans as a model organism.
Protocol

Biopharma Insights A protocol is described that uses anoxia/starvation in C. elegans to model ischemia/reperfusion. Functional outcomes include increased mortality, visible abnormalities in GFP-labeled neuronal processes, and impaired behavioral responses that require neuronal function.

Difficulty
intermediate
Total time
~4–6 hours (chamber setup ~1 hr; anoxia/starvation ~2–4 hrs; assessment ~1 hr)
Model organism
Caenorhabditis elegans
Biosafety
BSL-1

Steps

1
Construct a lab-made hypoxic chamber

Build a sealed anoxia chamber using standard laboratory materials to create oxygen-depleted conditions. The chamber design enables controlled exposure of C. elegans to anoxic stress.

▶ 01:55
2
Induce anoxia and starvation in C. elegans

Place worms in the hypoxic chamber under anoxic and starvation conditions for a defined period. This combined stress mimics ischemia/reperfusion-like injury in the nematode model.

▶ 03:06
3
Identify viable worms and assess touch response

Distinguish live from dead worms post-treatment and perform touch-response assays on survivors to measure behavioral recovery dependent on neuronal function.

▶ 05:53
4
Evaluate GFP-labeled neuronal morphology

Visualize and quantify structural abnormalities in GFP-tagged neuronal processes using fluorescence microscopy to assess neuronal damage from the anoxia/starvation insult.

▶ 06:38
5
Analyze survival and behavioral recovery data

Compile mortality rates and touch-response measurements post-anoxia/starvation to quantify functional and morphological outcomes of ischemia/reperfusion-like stress.

▶ 08:07
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