Characterization of Ca2+-dependent phospholipase A2 activity during zebrafish embryogenesis
- Farber, S.A., Olson, E.S., Clark, J.D., and Halpern, M.E.
- The Journal of biological chemistry 274(27): 19338-19346 (Journal)
- Registered Authors
- Farber, Steven, Halpern, Marnie E.
- MeSH Terms
- Arachidonic Acids/pharmacology
- Boron Compounds
- Enzyme Inhibitors/pharmacology
- Fluorescent Dyes
- Gene Expression Regulation, Developmental
- Gene Expression Regulation, Enzymologic
- Intracellular Membranes
- Models, Chemical
- Phospholipases A/antagonists & inhibitors
- Phospholipases A/genetics
- Phospholipases A/metabolism*
- Phospholipases A2
- 10383445 Full text @ J. Biol. Chem.
Farber, S.A., Olson, E.S., Clark, J.D., and Halpern, M.E. (1999) Characterization of Ca2+-dependent phospholipase A2 activity during zebrafish embryogenesis. The Journal of biological chemistry. 274(27):19338-19346.
We have developed a simple fluorescent assay for detection of phospholipase A2 (PLA2) activity in zebrafish embryos that utilizes a fluorescent phosphatidylcholine substrate. By using this assay in conjunction with selective PLA2 inhibitors and Western blot analysis, we identified the principal activity in zebrafish embryogenesis as characteristic of the Ca2+-dependent cytosolic PLA2 (cPLA2) subtype. Embryonic cPLA2 activity remained constant from the 1-cell stage until the onset of somitogenesis, at which time it increased sharply. This increase was preceded by the expression of a previously identified zebrafish cPLA2 homologue (Nalefski, E., Sultzman, L., Martin, D., Kriz, R., Towler, P., Knopf, J., and Clark, J. (1994) J. Biol. Chem. 269, 18239-18249). By using a quenched BODIPY-labeled phosphatidylcholine that fluoresces only upon cleavage by PLA2, lipase activity was visualized in the cells of living embryos where it localized to perinuclear membranes.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes