94
Methods for Setting Up Primary Cultures Specific to Animal Groups
Subprotocol 2
Parthenogenetic Embryos in Ovaries
Materials
Equipment - These are the same as given for subprotocoll
Ind solutions
Animals - Adults of parthenogenetic hemipteran insects, such as aphids
• • Procedure
In parthenogenetic aphids, embryos develop in the ovaries of their
mother.
Use viviparae adults containing many developing embryos. The developed red eye spots of the embryos can be seen through the cuticle of
their mother.
Sterilization l. Dip the aphids in 70% ethanol for 5 min.
2. Wash the aphids in sterile distilled water.
3. Transfer the aphids into a culture medium placed in the depression of
a Maximov slide.
Dissection l. Pinch the abdominal cuticle with two forceps. Tear the cuticle by pulling it with forceps (Fig. 3). Then, the ovaries containing developing
embryos will be exposed.
2. Remove the ovaries, taking care not to cut the intestine.
Culture l. Collect the ovaries, and cut the embryos into several pieces with
set-up
needles.
2. Transfer the embryo fragments into a culture vessel with the medium.
Comments
After the first step of culture set-up, the treatment described in the Comments section of subprotocol 1 may be applied .
• • Results
Aphid cells are extremely difficult to culture. Although cell migration
occurs to some extent (Fig. 4), only short survival of migrated cells has
been reported (Tokumitsu and Maramorosch, 1966; Peters and Black,
1971).
Methods for Setting Up Primary Cultures Specific to Animal Groups
Subprotocol 2
Parthenogenetic Embryos in Ovaries
Materials
Equipment - These are the same as given for subprotocoll
Ind solutions
Animals - Adults of parthenogenetic hemipteran insects, such as aphids
• • Procedure
In parthenogenetic aphids, embryos develop in the ovaries of their
mother.
Use viviparae adults containing many developing embryos. The developed red eye spots of the embryos can be seen through the cuticle of
their mother.
Sterilization l. Dip the aphids in 70% ethanol for 5 min.
2. Wash the aphids in sterile distilled water.
3. Transfer the aphids into a culture medium placed in the depression of
a Maximov slide.
Dissection l. Pinch the abdominal cuticle with two forceps. Tear the cuticle by pulling it with forceps (Fig. 3). Then, the ovaries containing developing
embryos will be exposed.
2. Remove the ovaries, taking care not to cut the intestine.
Culture l. Collect the ovaries, and cut the embryos into several pieces with
set-up
needles.
2. Transfer the embryo fragments into a culture vessel with the medium.
Comments
After the first step of culture set-up, the treatment described in the Comments section of subprotocol 1 may be applied .
• • Results
Aphid cells are extremely difficult to culture. Although cell migration
occurs to some extent (Fig. 4), only short survival of migrated cells has
been reported (Tokumitsu and Maramorosch, 1966; Peters and Black,
1971).