10 Insecta (Blattaria)
113
Fig. 4. Axonal outgrowth of cultured neurosecretory cells from a tenth ins tar nymph
of the American cockroach Periplaneta americana after 35 days in vitro. Reprinted
from Brain Research, Vo1.78, Seshan, K.R., Provine, R.R. and Levi-Montalcini, R.,
Structural and electrophysiological properties of nymphal and adult insect medial
neurosecretory cells: an in vitro analysis, 366,1974, with permission from Excerpta
Medica Inc.
- Forceps
- Needles
- Pasteur pipettes
- Maximov slides
- CO 2 incubator
- Scotch tape
- Insect Ringer's solution
- Culture medium (the same as that used in subprotocol 3)
- Nymphs or adults of P. americana
•
Procedure
1. Anesthetize the cockroach with carbon dioxide.
2. Fix the cockroach, dorsal side up, to a dissecting tray with scotch tape.
3. Expose the neck of the cockroach by applying light pressure to the
scotch tape across the thorax with the head of the cockroach tucked
inwards.
Solutions
Animals
Fixation
1. Sterilize the dorsal cervical sclerite with absorbent cotton moistened Sterilization
with 95% ethanol.
1. Make a slit in the region of the dorsal cervical sclerite and remove the Dissection
corpora cardiaca and corpora allata together using fine forceps (Fig. 5).
2. Transfer the excised organs into insect Ringer's solution placed in the
depression of a Maximov slide.
3. Remove adjoining tissues.
4. Wash the cleaned organs several times with Ringer's solution, and then
culture medium.