Home › Analytical Chem › Pharmacological and Functional Genetic Assays to Manipulate Regeneration of the Planarian Dugesia japonica
Analytical Chem JoVE (Open Access) Citable · DOI

Pharmacological and Functional Genetic Assays to Manipulate Regeneration of the Planarian Dugesia japonica

DOI: 10.3791/3058-v
What you'll learn
  • ✓ Perform amputation-based regeneration assays on planarian flatworms
  • ✓ Apply pharmacological agents to manipulate regeneration outcomes
  • ✓ Design and execute in vivo RNAi feeding protocols for genetic manipulation
Protocol

Biopharma Insights An attractive model for studying stem cell differentiation within a live animal is the planarian flatworm. Regeneration is studied by simple amputation experiments that are easily performed in a basic laboratory and are amenable to pharmacological and genetic (in vivo RNAi) manipulation as detailed by protocols in this article.

Difficulty
intermediate
Total time
~7–14 days (initial RNAi feeding phase ~5–7 days; regeneration observation ~7–14 days post-amputation)
Model organism
Planarian flatworm (Dugesia japonica)
Biosafety
BSL-1

Steps

1
Perform trunk fragment regeneration assay

Amputate planarian specimens to generate trunk fragments and establish baseline regeneration conditions. Monitor morphological outcomes over the observation period.

▶ 01:09
2
Induce bipolarity using praziquantel treatment

Apply praziquantel pharmacologically to manipulate regeneration polarity in amputated trunk fragments. Document phenotypic changes in head and tail formation.

▶ 03:00
3
Prepare and deliver in vivo RNAi by feeding

Generate dsRNA-loaded food pellets and feed planarians to achieve gene knockdown through oral RNAi. Maintain feeding schedule over multiple days.

▶ 04:13
4
Assess RNAi knockdown and regenerative phenotypes

Following RNAi treatment and amputation, evaluate regeneration outcomes and compare to control specimens. Document morphological and functional changes.

▶ 07:54
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