Chapter 28
Platyhelminthes
Subprotocol 1
Regeneration Blastema of Fresh-Water Plana ria (Turbellaria)
The following procedure is based on the methods of Betchaku (1970).
Materials
- Petri dish
- Pasteur pipettes
- Surgical knives
PROTOCOL
Equipment
- Saline solution (2,188 mg NaCI, 31 mg KCI, 63 mg CaClz, 125 mg Solutions
NaHC0 3 , and 100 mg neomycin sulfate in 1,000 ml distilled water,
pH 7.3)
- Wax block
Miscellaneous
- Freshwater Planaria, Dugesia dorotocephala (16-20-mm long)
Animals
Procedure
1. Clean the animals in autoclaved spring water with 0.02% neomycin Cleaning
sulfate with several changes of fresh solution over a period of 10-24 h. animals
1. Cut out a cross-piece of the animal midway between the head and the Dissection
pharynx in saline solution.
1. Slice the cut pieces on a wax block, dorso-ventrally, into small tissue Culture
cubes of between 0.1 and 0.03 mm 3 •
set-up
2. Keep these fragments undisturbed in saline solution at 17°-18°C.
Results
According to Betchaku (1970), the tissue fragments remained essentially
motionless, with slight occasional undulation, and maintained their original cubic form for the first few hours. Morphologic rearrangement of
tissues was completed in 2 days. Parenchyma accounted for most of the
motility of the inner composite tissue layer. Regeneration blastemas appeared to form by local cellular segregation triggered by injury under
hypotonic conditions.
Platyhelminthes
Subprotocol 1
Regeneration Blastema of Fresh-Water Plana ria (Turbellaria)
The following procedure is based on the methods of Betchaku (1970).
Materials
- Petri dish
- Pasteur pipettes
- Surgical knives
PROTOCOL
Equipment
- Saline solution (2,188 mg NaCI, 31 mg KCI, 63 mg CaClz, 125 mg Solutions
NaHC0 3 , and 100 mg neomycin sulfate in 1,000 ml distilled water,
pH 7.3)
- Wax block
Miscellaneous
- Freshwater Planaria, Dugesia dorotocephala (16-20-mm long)
Animals
Procedure
1. Clean the animals in autoclaved spring water with 0.02% neomycin Cleaning
sulfate with several changes of fresh solution over a period of 10-24 h. animals
1. Cut out a cross-piece of the animal midway between the head and the Dissection
pharynx in saline solution.
1. Slice the cut pieces on a wax block, dorso-ventrally, into small tissue Culture
cubes of between 0.1 and 0.03 mm 3 •
set-up
2. Keep these fragments undisturbed in saline solution at 17°-18°C.
Results
According to Betchaku (1970), the tissue fragments remained essentially
motionless, with slight occasional undulation, and maintained their original cubic form for the first few hours. Morphologic rearrangement of
tissues was completed in 2 days. Parenchyma accounted for most of the
motility of the inner composite tissue layer. Regeneration blastemas appeared to form by local cellular segregation triggered by injury under
hypotonic conditions.