Wong Julian L, Créton Robbert, Wessel Gary M
Department of Molecular Biology, Cell Biology, and Biochemistry, Box G, Brown University, Providence, RI 02912 USA.
Dev Cell. 2004 Dec;7(6):801-14. doi: 10.1016/j.devcel.2004.10.014.
The sea urchin egg is a quiescent cell...until fertilization, when the egg is activated. The classic respiratory burst at fertilization is the result of prodigious hydrogen peroxide production, but the mechanism for this synthesis is not known. Here we quantitate the kinetics of hydrogen peroxide synthesis at a single-cell level using an imaging photon detector, showing that 60 nM hydrogen peroxide accumulates within the perivitelline space of each zygote. We find that the NADPH oxidation activity is enriched at the cell surface and is sensitive to a pharmacological inhibitor of NADPH oxidase enzymes. Finally, we show that a sea urchin dual oxidase homolog, Udx1, is responsible for generating the hydrogen peroxide necessary for the physical block to polyspermy. Phylogenetic analysis of the enzymatic modules in Udx1 suggests a potentially conserved role for the dual oxidase family in hydrogen peroxide production and regulation during fertilization.
海胆卵是一种静止细胞……直到受精时,卵子被激活。受精时经典的呼吸爆发是大量过氧化氢产生的结果,但这种合成的机制尚不清楚。在这里,我们使用成像光子探测器在单细胞水平上定量过氧化氢合成的动力学,结果表明每个受精卵的卵周隙内积累了60 nM的过氧化氢。我们发现NADPH氧化活性在细胞表面富集,并且对NADPH氧化酶的药理抑制剂敏感。最后,我们表明海胆双氧化酶同源物Udx1负责产生对多精入卵进行物理阻断所必需的过氧化氢。对Udx1中酶模块的系统发育分析表明,双氧化酶家族在受精过程中过氧化氢的产生和调节中可能具有保守作用。