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Cell Biology JoVE (Open Access) Citable · DOI

Methods to Assess Subcellular Compartments of Muscle in C. elegans

DOI: 10.3791/52043-v
What you'll learn
  • ✓ Synchronize C. elegans L1 larvae using gravity flotation technique
  • ✓ Assess muscle function via movement assays and locomotion analysis
  • ✓ Visualize mitochondrial networks and sarcomere structure using GFP imaging
  • ✓ Measure muscle protein degradation and membrane potential in body wall muscle
Protocol

Skeletal muscle is essential for locomotion and is the bodies’ main protein store. Muscle health measurements within C. elegans are described. Prospective changes to muscle structure and function are assessed using localized GFP and cationic dyes.

Difficulty
intermediate
Total time
~3–4 hours per experimental cohort (including larval synchronization, imaging, and assays)
Model organism
Caenorhabditis elegans
Biosafety
BSL-1

Steps

1
Synchronize L1 larvae using gravity flotation

Use gravity flotation method to obtain synchronized L1 larval C. elegans populations for standardized downstream muscle assays.

▶ 01:50
2
Perform movement assays on animals

Measure locomotion and movement capacity as a functional readout of muscle health in synchronized worms.

▶ 03:08
3
Image mitochondrial networks and sarcomeres

Use GFP-based imaging to visualize mitochondrial organization and sarcomeric structure in body wall muscle; assess membrane potential with cationic dyes.

▶ 03:59
4
Assess muscle protein degradation levels

Quantify protein degradation in muscle tissue using established fluorescent or biochemical methods to evaluate catabolic state.

▶ 06:13
5
Analyze effects of muscle disruption

Interpret combined data from movement, imaging, and protein degradation assays to evaluate phenotypic consequences of muscle perturbation.

▶ 07:57
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