10 Insecta (Biattaria)
III
5. Assemble five to six culture vessels in a petri dish in which absorbent
cotton soaked with sterile distilled water has been placed.
6. Keep the petri dish in a desiccator filled with 95% air and 5% CO 2 •
7. Culture at 29°C.
Results
According to Chen and Levi-Montalcini (1970), brain cells survive this
culture process, but glial cells do not. The pure nerve cell population was
maintained for as long as 4-5 months in the presence of foregut explants
from the same embryo, whereas, if cultured alone, the nerve cell population survived for some weeks but did not produce a fibrillar network.
Radial fiber bundles emerging from the nerve fiber networks converged
toward the explant and established synaptic connections with it (Fig. 3).
Subprotocol 4
Neurosecretory Cells
The following method is based on that reported by Seshan and LeviMontalcini (1973).
Materials
- Dissecting microscope
- Dissecting tray (see chapter 1, section 3 "Tools")
- Maximov slides
- Scissors
- Forceps
- Needles
- Pasteur pipettes
- Physiological solution
- Culture medium (the same as that used in subprotocol3)
- Nymphs or adults of P. americana
Procedure
Equipment
Solutions
Animals
1. Surface sterilize the cockroaches by submersion in 70% ethanol for Sterilization
5 min.
1. Open the head capsule, and remove the whole brain.
Dissection
2. Place the brain in a physiological solution in the depression of a
Maximov slide. The two groups of medial neurosecretory cells can be
identified in the intact brain under a dissecting microscope by their
position, as well as by the pale bluish color, due to light scattering
III
5. Assemble five to six culture vessels in a petri dish in which absorbent
cotton soaked with sterile distilled water has been placed.
6. Keep the petri dish in a desiccator filled with 95% air and 5% CO 2 •
7. Culture at 29°C.
Results
According to Chen and Levi-Montalcini (1970), brain cells survive this
culture process, but glial cells do not. The pure nerve cell population was
maintained for as long as 4-5 months in the presence of foregut explants
from the same embryo, whereas, if cultured alone, the nerve cell population survived for some weeks but did not produce a fibrillar network.
Radial fiber bundles emerging from the nerve fiber networks converged
toward the explant and established synaptic connections with it (Fig. 3).
Subprotocol 4
Neurosecretory Cells
The following method is based on that reported by Seshan and LeviMontalcini (1973).
Materials
- Dissecting microscope
- Dissecting tray (see chapter 1, section 3 "Tools")
- Maximov slides
- Scissors
- Forceps
- Needles
- Pasteur pipettes
- Physiological solution
- Culture medium (the same as that used in subprotocol3)
- Nymphs or adults of P. americana
Procedure
Equipment
Solutions
Animals
1. Surface sterilize the cockroaches by submersion in 70% ethanol for Sterilization
5 min.
1. Open the head capsule, and remove the whole brain.
Dissection
2. Place the brain in a physiological solution in the depression of a
Maximov slide. The two groups of medial neurosecretory cells can be
identified in the intact brain under a dissecting microscope by their
position, as well as by the pale bluish color, due to light scattering