This experiment is suitable for students at any level of education. The aim of this experiment is to observe the regeneration of worms following amputation to identify: blastema formation, antero-posterior (A/P) and dorsal-ventral (D/V) axes conservation, eye regrowth, re-pigmentation of the regenerated tissues, and worm behavior. In Section 2.1, three different amputation paradigms are suggested, but the student should feel free to experiment also different strategies.
REQUIRED MATERIALS |
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Plastic Pasteur pipettes |
Petri dishes |
Ethanol |
Blade or coverslip |
Paper towels or lab wiper KimWipes™ |
Optional: Lens magnification or dissecting microscope or smartphone |
Squeeze bottle or standard bottle and plastic Pasteur pipettes |
Instant Ocean® Sea Salt or 1x Montjuïc water |
Waste container |
Put the desired number of 7-day starved worms in a Petri dish of adequate size as reported in Section 1.1 on the basis of the final number of fragments to be obtained.
The suggested amputation strategies are:
REQUIRED MATERIALS |
---|
Lens magnification or dissecting microscope or smartphone |
Optional: Camera |
Plastic Pasteur pipettes |
Petri dishes |
Squeeze bottle or standard bottle and plastic Pasteur pipettes |
Cold Instant Ocean® Sea Salt or 1x Montjuïc water |
Waste container |
During amputation students should pay attention to the “escape” behavior of the worms, mucous production, and body contraction. During the days after the amputation, the students can examine wound closure by muscle contraction, blastema formation, swimming behavior, conservation of the body axes, tissue remodeling to re-establish proper body proportions, eye formation, and pigmentation of the regenerated tissues that represents the end of the process.
A detailed description of the regeneration time course of each planarian species after each amputation paradigm are available in the paper “Hands-on, classroom studies of regeneration, and stem cell biology using freshwater planarians”.
The specific features of each observed time point can be documented with picture acquisition and movie recording as described in the Section 1.5.