21 Insecta
249
- Pasteur pipettes
- Petri dishes
- Maximov slide
- Culture vessels
- Physiological solution
Solutions
- Culture media
- Pupae of lepidopteran insects
Animals
Procedure
1. Soak the pupae in 70 % ethanol for 5 min.
Sterilization
2. Rinse the pupae with sterilized distilled water.
1. Fix the pupae using insect pins on a dissecting tray filled with physi- Dissection
ological solution.
2. Make an incision on the anterior part of the pupa.
3. Excise the brain carefully so as to avoid injuring the gut.
1. Wash the brain in physiological solution placed in a depression of a Culture
Maximov slide, and remove any attached tissues.
set-up
2. Transfer the brain into a culture vessel with culture medium.
Results
Leloup and Marks (1973) cultured the brains of diapausing pupae of the
tobacco horn worm Manduca sexta in a modified Grace's medium (see
Appendix 1). During cultivation, mitosis was observed in the cells of optic lobes, and neurosecretory materials moved to the corpora allata. Agui
and Hiruma (1977) cultured the brains of diapausing pupae of the cabbage armyworm M. brassicae in CSM-2F medium (see Appendix O. The
synthesis of neurosecretory materials by neurosecretory cells of the pars
intercerebralis seemed to be increased following the addition of 20hydroxyecdysone to the culture medium.
Subprotocol 8
Corpora Cardiaca-Corpora Allata Complex of Nymphal
Cockroaches (Blattaria)
Materials
- Dissecting microscope
- Maximov slide
- Scotch tape
- Fine forceps
Equipment
249
- Pasteur pipettes
- Petri dishes
- Maximov slide
- Culture vessels
- Physiological solution
Solutions
- Culture media
- Pupae of lepidopteran insects
Animals
Procedure
1. Soak the pupae in 70 % ethanol for 5 min.
Sterilization
2. Rinse the pupae with sterilized distilled water.
1. Fix the pupae using insect pins on a dissecting tray filled with physi- Dissection
ological solution.
2. Make an incision on the anterior part of the pupa.
3. Excise the brain carefully so as to avoid injuring the gut.
1. Wash the brain in physiological solution placed in a depression of a Culture
Maximov slide, and remove any attached tissues.
set-up
2. Transfer the brain into a culture vessel with culture medium.
Results
Leloup and Marks (1973) cultured the brains of diapausing pupae of the
tobacco horn worm Manduca sexta in a modified Grace's medium (see
Appendix 1). During cultivation, mitosis was observed in the cells of optic lobes, and neurosecretory materials moved to the corpora allata. Agui
and Hiruma (1977) cultured the brains of diapausing pupae of the cabbage armyworm M. brassicae in CSM-2F medium (see Appendix O. The
synthesis of neurosecretory materials by neurosecretory cells of the pars
intercerebralis seemed to be increased following the addition of 20hydroxyecdysone to the culture medium.
Subprotocol 8
Corpora Cardiaca-Corpora Allata Complex of Nymphal
Cockroaches (Blattaria)
Materials
- Dissecting microscope
- Maximov slide
- Scotch tape
- Fine forceps
Equipment