ZFIN ID: ZDB-PUB-990628-5
Characterization of Ca2+-dependent phospholipase A2 activity during zebrafish embryogenesis
Farber, S.A., Olson, E.S., Clark, J.D., and Halpern, M.E.
Date: 1999
Source: The Journal of biological chemistry   274(27): 19338-19346 (Journal)
Registered Authors: Farber, Steven, Halpern, Marnie E.
Keywords: none
MeSH Terms:
  • Animals
  • Arachidonic Acids/pharmacology
  • Blastoderm/enzymology
  • Boron Compounds
  • Calcium/metabolism*
  • Enzyme Inhibitors/pharmacology
  • Female
  • Fluorescent Dyes
  • Gastrula/enzymology
  • Gene Expression Regulation, Developmental
  • Gene Expression Regulation, Enzymologic
  • Intracellular Membranes
  • Male
  • Models, Chemical
  • Organophosphonates/pharmacology
  • Phospholipases A/antagonists & inhibitors
  • Phospholipases A/genetics
  • Phospholipases A/metabolism*
  • Phospholipases A2
  • Zebrafish/embryology*
PubMed: 10383445 Full text @ J. Biol. Chem.
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.