Protocol
Protocol for Drosophila ovary dissection
- Feed flies with yeast 1-2 days prior to dissecting them to fatten up the ovaries.
- Anaesthetize flies using carbon dioxide stream.
- Using a pair of tweezers, select a female fly.
- Submerge the female fly into 1X PBS.
- Grab the fly at its lower thorax with a pair of tweezers.
- Tug gently at the lower abdomen with another pair of tweezers until the internal organs in the abdomen are exposed.
- Look for the pair of ovaries and detach it from other organs (e.g. the intestines).
- Tease apart the ovarioles (if ovaries are to be used for immunostaining or in situ hybridization).
- Keep ovaries in ice-cold 1X PBS while dissecting the next fly.
Discussion
Here we demonstrate a method of dissecting ovaries in Drosophila. As the method of culturing the different cell types that exist in the Drosophila ovary has yet been defined, this is a rapid method to obtain the Drosophila ovarian tissue. Essentially, we show that dissecting Drosophila ovaries involves a simple two- or three-step procedure. Subsequent treatment of these dissected ovaries depend on the downstream experiments that will be performed.
Materials
| Material Name | Tipo | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| Forceps | Tool | Sigma | T43537 | |
| Glass dissecting dish | Tool | Pyrex | 7220-85 |
Referências
- Costa, A., et al. The Drosophila fragile X protein functions as a negative regulator in the orb autoregulatory pathway. Dev Cell. 8, 331-342 (2005).
- Tan, L., et al. An autoregulatory feedback loop directs the localized expression of the Drosophila CPEB protein Orb in the developing oocyte. Development. 128, 1159-1169 (2001).
Citar este artigo
Wong, L. C., Schedl, P. Dissection of Drosophila Ovaries. J. Vis. Exp. (1), e52, doi:10.3791/52 (2006).