Acetylcholinesterase activity in the cornea of the developing chick embryo.
Activity of acetylcholinesterase (EC 3.1.1.7) and pseudocholinesterase (EC 3.1.1.8) was measured in extracts from chick corneas, in a developmental series from days 7-20 of incubation and at three ages after hatching. Enzyme activity was measured by the biphasic single-vial radiometric assay of Johnson and Russell using [3H-acetyl]choline as substrate. Pseudocholinesterase was inhibited with tetraisopropylpyrophosphoramide (iso-OMPA). True acetylcholinesterase activity was verified by control assays run in the presence of both iso-OMPA and the true acetylcholinesterase inhibitor, 1:5-bis(4-allyldimethyl-ammonium phenyl)-pentane-3-one diiodide (BW284c51). With both inhibitors present, no cholinesterase activity was detected. Corneal acetylcholinesterase had an average Km of 1.1 +/- 0.3 X 10(-3) M at day 7, 14, and 20 of development and retained 90% activity even after 3 hr at 26 degrees C. At least 90% of the total cholinesterase activity was solubilized by Triton X-100 and sonication treatment. Activity decreased with increasing concentrations of NaCl present in the assay. A 60-fold transient increase in acetylcholinesterase specific activity occurs during the period from days 7-20 of embryonic development. This increase begins on the first day measured (day 7), progresses steadily and rapidly during the subsequent week, reaches a peak at day 15, and then decreases rapidly before hatching to a level maintained into adulthood. A similar pattern of transient appearance of highly sialylated gangliosides seen previously on days 14-17 leads to an hypothesis of a structural linkage between acetylcholinesterase and the plasma membrane lipids of corneal epithelial cells.
D000110:Acetylcholinesterase / Q000378:metabolism*; D000818:Animals; D002091:Butyrylcholinesterase / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002642:Chick Embryo; D002802:Cholinesterases / Q000378:metabolism*; D003315:Cornea / Q000196:embryology* / Q000201:enzymology; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D008563:Membrane Lipids / Q000378:metabolism
D008563:Membrane Lipids; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase; D002802:Cholinesterases