PUBLICATION
Characterization and In Vitro Control of MPF Activity in Zebrafish Eggs
- Authors
- Siripattarapravat, K., Busta, A., Steibel, J.P., and Cibelli, J.
- ID
- ZDB-PUB-090319-16
- Date
- 2009
- Source
- Zebrafish 6(1): 97-105 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Animals
- Cellular Senescence
- Female
- Fertilization
- Maturation-Promoting Factor/metabolism*
- Oocytes/metabolism*
- Zebrafish/metabolism*
- PubMed
- 19292671 Full text @ Zebrafish
Citation
Siripattarapravat, K., Busta, A., Steibel, J.P., and Cibelli, J. (2009) Characterization and In Vitro Control of MPF Activity in Zebrafish Eggs. Zebrafish. 6(1):97-105.
Abstract
We describe the characterization of maturation-promoting factor (MPF) in zebrafish eggs and used different defined conditions to maintain its activity in vitro. MPF activity levels are high in freshly ovulated mature eggs and decline rapidly within 5 min after either fertilization or parthenogenetic activation. The MPF activity of eggs matured in vitro declines faster when the eggs are incubated in Hank's culture medium supplemented with 0.5% BSA (H-BSA) than when incubated in Chinook salmon ovarian fluid (CSOF). MPF activity in nonactivated, aged eggs remains high in H-BSA supplemented with 75 muM MG132 or 10 mM caffeine, but neither MG132 nor caffeine can sustain high MPF activity in activated eggs. MG132-treated eggs showed delayed completion of metaphase and extrusion of the second polar body. Nuclear staining of the activated eggs confirmed the correlation between their cell cycle stage and MPF activity at each time point. An embryotoxic effect was found when matured eggs were held in 100 muM of MG132 or 20 mM caffeine for 1 h. Calcium-depleted medium and 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid also showed detrimental effects on the embryos. Conversely, nonactivated, aged matured eggs maintained high MPF activity and developmental potential when CSOF was used as a holding medium.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping