Lipid Signaling During Fertilization

受精过程中的脂质信号传导

基本信息

批准号：
7939130
负责人：
BRADLEY J STITH
金额：
$ 44.99万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2013-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7939130
关键词：
Acids Acrosome Reaction Affinity Binding Biological Assay Biological Models Biology Calcium Colorado Cytoplasmic Granules Data Defect Detection Development Dominant-Negative Mutation Equipment Event Exocytosis Fertilization Funding Future Goals Human Infertility Inositol Kentucky Lead Letters Lipase Lipids Lysophospholipids Manuscripts Mass Spectrum Analysis Measures Membrane Membrane Fusion Membrane Lipids Methodology Methods Modeling Molecular Phosphatidic Acid Phospholipase C Phospholipase D Phospholipid Metabolism Production Protein Kinase C Protocols documentation Publications Publishing Reporting Research Role Series Signal Pathway Signal Transduction Surveys System Techniques Universities Work Xenopus Xenopus laevis Xenopus oocyte base cellular microvillus egg experience insight light scattering lysophosphatidic acid mass spectrometer medical schools novel programs public health relevance sperm cell

项目摘要

DESCRIPTION (provided by applicant): There is numerous membrane events associated with fertilization: phospholipids metabolism, membrane fusion during the sperm acrosome reaction, during sperm-egg fusion, during cortical granule exocytosis, and changes in microvilli. Surprisingly, only phospholipase C activation (PLC)(which leads to production of inositol 1,4,5- trisphosphate and sn 1,2-DAG, calcium release and protein kinase C activation, has been widely studied. Over the 15 years, my lab has used specialized lipid analysis equipment and protocols developed with Avanti Polar Lipids to undertake a unique series of studies on other lipid signaling pathways. Our recent publication (selected over other articles by ASBMB Today) suggests roles for phospholipase D and autotaxin in fertilization in an NIH-recognized model system (Xenopus laevis). Our long term goal is obtain a survey of lipid signaling paths activated during fertilization and detail their role in the induction of fertilization events. We will inhibit autotaxin activity, quantify the levels of lysophosphatidic acid (measured by HPLC/ELSD or mass spectrometry) and examine the effect upon fertilization events in Xenopus. In the last two specific aims, we will examine phospholipase D (which catalyzes phosphatidic acid formation) during fertilization. We also provide extensive Preliminary Results demonstrating that increased phosphatidic acid (PA) production is involved in fertilization events such as the elevation of DAG, the activation of Src, PLC, and elevation of IP3 and intracellular calcium. We will use proven methods to quantify (Kd) the affinity between LPA, PA and Src. We will alter lipase D activity (by use of constitutively active or dominant negative PLD) or morpholinos against Xenopus phospholipase D message. These studies will make use of our expertise and specialized equipment in lipid signaling and Xenopus biology obtained over the past 20 years. PUBLIC HEALTH RELEVANCE: Although there are many lipid events during fertilization, they have not been well studied. Funding of this proposal will support the first study on the role of autotaxin and phospholipase D in fertilization. This work should provide insight into the mechanism of fertilization and may lead to identification of defects associated with human infertility.

描述（由申请人提供）：有许多与受精有关的膜事件：磷脂代谢，精子质体反应期间的膜融合，精子 - 蛋性融合期间，皮质颗粒胞外生殖过程和微伏的变化。出乎意料的是，仅研究了1,4,5-磷酸肌醇1,4,5-磷酸和Sn 1,2-DAG，钙释放和蛋白激酶C激活的磷酸酶C活化（PLC）（PLC）。在15年中，我已经在15年中进行了广泛的研究。在这15年中，我的LAB最近在Avanti Polarworke上开发了一个独特的LIPID Seriess in lip Lip-Lip-Lip-Lip-nequare Serecters系列。（当今ASBMB选择其他文章）表明，在NIH认识的模型系统中，磷脂酶D和自身X。 HPLC/ELSD或质谱法）在最后两个特定目标中检查对Xenopus的施肥事件的影响，我们将检查磷脂酶D（在受精过程中催化磷脂酸）。我们还提供了广泛的初步结果，表明磷脂酸（PA）的产生增加参与受精事件，例如DAG升高，SRC，PLC的激活以及IP3的升高和细胞内钙的升高。我们将使用经过验证的方法来量化LPA，PA和SRC之间的亲和力。我们将改变脂肪酶D活性（通过使用组成型活性或显性阴性PLD）或形态上的磷脂酶D消息。这些研究将利用我们在过去20年中获得的脂质信号传导和Xenopus生物学方面的专业知识和专业设备。公共卫生相关性：尽管受精过程中有许多脂质事件，但研究并未得到很好的研究。该提案的资金将支持第一项关于自身蛋白酶和磷脂酶D在受精中的作用的研究。这项工作应提供对受精机制的见解，并可能导致识别与人类不育相关的缺陷。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Increasing pro-survival factors within whole brain tissue of Sprague Dawley rats via intracerebral administration of modified valproic acid.

DOI：
10.1016/j.jphs.2015.07.003
发表时间：
2015-08
期刊：
Journal of pharmacological sciences
影响因子：
3.5
作者：
R. Bates;B. Stith;K. Stevens
通讯作者：
R. Bates;B. Stith;K. Stevens

Activation of Src and release of intracellular calcium by phosphatidic acid during Xenopus laevis fertilization.

DOI：
10.1016/j.ydbio.2013.11.006
发表时间：
2014-02-01
期刊：
DEVELOPMENTAL BIOLOGY
影响因子：
2.7
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.
通讯作者：
Stith, Bradley J.

Insemination or phosphatidic acid induces an outwardly spiraling disk of elevated Ca2+ to produce the Ca2+ wave during Xenopus laevis fertilization.

授精或磷脂酸会诱导Ca2+向外螺旋盘升高，从而在非洲爪蟾受精过程中产生Ca2+波。