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punctata, and showed juvenile hormone synthesis during short-and longterm cultures. The performance of the gland was improved when L-15B
medium (see Appendix 1) was used and when the gland was floating at
the surface of the medium.
Subprotocol 9
Cephalic Complex of Flies (Diptera)
Materials
- Dissecting microscope
- Petri dishes
- Maximov slide
- Scissors
- Forceps
- Needles
- Pasteur pipettes
- Culture vessels
- Physiological solution
- Culture medium (Leloup's medium 1; see Appendix 1)
- Prepupae of the blowfly Calliphora erythrocephala
Procedure
Equipment
Solutions
Animals
1. Surface sterilize the prepupae of flies by submersion in 70% ethanol Sterilization
for 5 min.
2. Rinse the prepupae with sterilized distilled water.
1. Soak the prepupae in physiological solution placed in the depression Dissection
of a Maximov slide.
2. Excise the brains with the ring glands from prepupae, care being taken
not to damage the alimentary tract.
1. Remove the tissues attached to the cephalic complex in physiological Culture
solution.
set-up
2. Transfer the complex into a culture vessel with culture medium.
3. Culture the glands at 25°C.
Results
According to Leloup and Gianfelici (1966), the cephalic complex of the
blowfly could be maintained in culture for more than 4 days in a healthy
condition. The median neurosecretory cells of the brain showed positive
staining for secretory material even after 4 days culture.