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Analysis of the Regulatory Mechanism and Function of a Novel Class of Phospholipase C, PLCε

一类新型磷脂酶 C PLCε 的调控机制和功能分析

基本信息

批准号：
15390093
负责人：
KATAOKA Tohru
金额：
$ 9.86万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390093/
关键词：
phospholipase C small G protein Ras protein phosphoinositide signaling ventricular dilation semilunar valve disease knockout mice nematodes Rap1蛋白質半月弁逆流半月弁狭窄

项目摘要

1, We constructed phospholipase C_ε(PLC_ε) knockout mice and showed that the hearts of these mice develop ventricular dilation, which is caused by a volume overload resulting from marked regurgitation and mild stenosis of the semilunar (aortic and pulmonic) valves. We analysed the mechanism of the congenital semilunar valvulogenesis defect causing these phenotypes and concluded that the abnormal thickening and morphology of the semilunar valve leaflets in PLC_ε knockout mice were caused by aberrant cellular proliferation in valve remodeling at the late stages of semilunar valvulogenesis. By analogy with the phenotypes of mice carrying an attenuated epidermal growth factor (EGF) receptor or deficient in heparin-binding EGF, we speculated a crucial role of PLC_ε, as a Ras effector downstream of the EGF receptor, in inhibition of valvular cell proliferation. In fact, we observed increased phosphorylation of Smad1/5/8, which induce valvular cell proliferation downstream of the BMP receptor, in PLC_ε knockout mice.2, By applying the two stage skin chemical carcinogenesis protocol using DMBA as an initiator and a phorbor ester TPA as a promoter to PLC_ε knockout mice, we showed that PLC_ε plays a crucial role in ras oncogene-induced development of benign tumors as well as in their malignant progression. We further analysed the mechanism of action of PLC_ε. PLC_ε knockout mice showed marked reduction in proliferation of basal layer cells and epidermal hyperplasia induced by TPA, suggesting a crucial role of PLC_ε in downstream signaling from TPA.3, We disrupted the PLC_ε gene in a model organism Caenorhabditis elegans and observed a sterile phenotype, caused by abnormal contraction of sphincter muscles of the spermatheca.4, By using BaF3 cells expressing a PDGF receptor mutant lacking the PLC_γ-binding sites, we showed the importance of PH and RA domains by establishing an assay system for Ca^<2+> increase induced by PDGF-dependent PLC_ε activation.

1、我们构建了磷脂酶C_ε（PLC_ε）基因敲除小鼠模型，发现其心脏出现心室扩张，这是由明显的反流和半月瓣（主动脉瓣和肺动脉瓣）轻度狭窄引起的容量超负荷引起的。我们分析了先天性半月瓣发育缺陷导致这些表型的机制，认为PLC_ε基因敲除小鼠半月瓣小叶的异常增厚和形态是由半月瓣发育后期瓣膜重塑中的异常细胞增殖引起的。通过与携带减弱的表皮生长因子（EGF）受体或缺乏肝素结合的EGF的小鼠的表型相类比，我们推测PLC_ε作为EGF受体下游的Ras效应子在抑制瓣膜细胞增殖中的关键作用。事实上，我们观察到在PLC_ε基因敲除小鼠中，Smad 1/5/8的磷酸化增加，从而诱导BMP受体下游的瓣膜细胞增殖。2.通过应用以DMBA为引发剂和以佛波酯TPA为促进剂的两阶段皮肤化学致癌方案，我们发现PLC_ε在ras癌基因诱导的良性肿瘤的发展以及恶性进展中起着至关重要的作用。并进一步分析了PLC_ε的作用机理。PLC_ε基因敲除小鼠的睾丸基底层细胞增殖和表皮增生明显减少，表明PLC_ε在TPA下游信号传导中起重要作用。3、我们在模式生物秀丽隐杆线虫中敲除PLC_ε基因，观察到受精囊括约肌异常收缩导致的不育表型。利用表达缺少PLC_γ结合位点的PDGF受体突变体的BaF 3细胞，我们通过建立一个检测系统，检测PDGF依赖的PLC_ε激活诱导的Ca^<2+>增加，从而证明了PH和RA结构域的重要性。

项目成果

期刊论文数量（52）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Role of the Sec14-like domain of DbI family exchange factors in the regulation of Rho family GTPases in different subcellular sites

DbI家族交换因子的Sec14样结构域在不同亚细胞位点Rho家族GTP酶调节中的作用

DOI：
发表时间：
2004
期刊：
Cellular Signalling 16巻・8号
影响因子：
0
作者：
Shuji Ueda;et al.
通讯作者：
et al.

Neuronal lineage‐specific induction of phospholipase Cε expression in the developing mouse brain

DOI：
10.1046/j.1460-9568.2003.02591.x
发表时间：
2003-04
期刊：
European Journal of Neuroscience
影响因子：
3.4
作者：
Dongmei Wu;M. Tadano;H. Edamatsu;Misa Masago-Toda;Y. Yamawaki‐Kataoka;T. Terashima;A. Mizoguchi
通讯作者：
Dongmei Wu;M. Tadano;H. Edamatsu;Misa Masago-Toda;Y. Yamawaki‐Kataoka;T. Terashima;A. Mizoguchi

Phospholipase Cε regulates ovulation in Caenorhabditis elegans