Activation of Src and release of intracellular calcium by phosphatidic acid during Xenopus laevis fertilization.
Activation of Src and release of intracellular calcium by phosphatidic acid during Xenopus laevis fertilization.
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DOI：
10.1016/j.ydbio.2013.11.006
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发表时间：
2014-02-01
影响因子：
2.7
通讯作者：
Stith, Bradley J.
中科院分区：
文献类型：
--
作者：
Bates, Ryan C.;Fees, Colby P.;Holland, William L.;Winger, Courtney C.;Batbayar, Khulan;Ancar, Rachel;Bergren, Todd;Petcoff, Douglas;Stith, Bradley J.
我们报道了非洲爪蟾受精的一个新步骤，该步骤涉及激活Src酪氨酸激酶以刺激磷脂酶C-γ (PLC- γ)，从而增加肌醇1,4,5-三磷酸（IP3）以释放细胞内钙（[Ca]i）。分子种类分析和质量测量表明，精子激活磷脂酶D （PLD）提高磷脂酸（PA）。我们现在报道，在受精后1分钟内，PA质量增加了2.7倍，通过两种方法抑制PA的产生抑制了Src和PLCγ的激活，增加了[Ca]i和其他受精事件。与其他14种脂质相比，PA与爪蟾Src结合较强，而与PLCγ不结合。添加合成PA激活了鸡蛋Src（需要完整的脂筏）和PLCγ，并使筏中PLCγ的数量增加一倍。在没有升高[Ca]i的情况下，PA的加入将IP3质量提高到与精子诱导的水平相当（但是钙离子载体所达到的水平的两倍）。最后，PA诱导的[Ca]i释放被IP3受体抑制剂阻断。由于仅检测到PLD1b信息，而Western blotting未检测到PLD2，我们认为精子激活PLD1b以升高PA， PA随后结合并激活Src，导致PLCγ刺激，IP3升高和[Ca]i释放。由于这些和其他研究，PA也可能在膜融合事件中发挥作用，如精子-卵子融合、皮质颗粒胞吐、磷脂酰肌醇4,5-二磷酸的升高以及受精过程中sn 1,2-二酰基甘油的大量晚期增加。
We report a new step in the fertilization in Xenopus laevis which has been found to involve activation of Src tyrosine kinase to stimulate phospholipase C-γ (PLC- γ) which increases inositol 1,4,5-trisphosphate (IP3) to release intracellular calcium ([Ca]i). Molecular species analysis and mass measurements suggested that sperm activate phospholipase D (PLD) to elevate phosphatidic acid (PA). We now report that PA mass increased 2.7 fold by 1 minute after insemination and inhibition of PA production by two methods inhibited activation of Src and PLCγ, increased [Ca]i and other fertilization events. As compared to 14 other lipids, PA strongly bound Xenopus Src but not PLCγ. Addition of synthetic PA activated egg Src (an action requiring intact lipid rafts) and PLCγ as well as doubling the amount of PLCγ in rafts. In the absence of elevated [Ca]i, PA addition elevated IP3 mass to levels equivalent to that induced by sperm (but twice that achieved by calcium ionophore). Finally, PA induced [Ca]i release that was blocked by an IP3 receptor inhibitor. As only PLD1b message was detected, and Western blotting did not detect PLD2, we suggest that sperm activate PLD1b to elevate PA which then binds to and activates Src leading to PLCγ stimulation, IP3 elevation and [Ca]i release. Due to these and other studies, PA may also play a role in membrane fusion events such as sperm-egg fusion, cortical granule exocytosis, the elevation of phosphatidylinositol 4,5-bisphosphate and the large, late increase in sn 1,2-diacylglycerol in fertilization.
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影响因子：
4.4
作者：
Chalifa-Caspi, V;Eli, Y;Liscovitch, M
通讯作者：
Liscovitch, M
影响因子：
5.8
作者：
Chang, Ling-Chu;Huang, Tai-Hung;Wang, Jih-Pyang
通讯作者：
Wang, Jih-Pyang
影响因子：
5.8
作者：
Faugaret, Delphine;Chouinard, Francois C.;Bourgoin, Sylvain G.
通讯作者：
Bourgoin, Sylvain G.
影响因子：
--
作者：
DURIEUX, ME;SALAFRANCA, MN;MOORMAN, JR
通讯作者：
MOORMAN, JR
影响因子：
3.5
作者：
Exton, JH
通讯作者：
Exton, JH