Elucidation of the synapse formation mechanism with glutamatergic signaling

通过谷氨酸信号阐明突触形成机制

基本信息

批准号：
16500192
负责人：
TSUZUKI Keisuke
金额：
$ 2.37万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

AMPA-type glutamate receptors were composed of four subunits, GluR1, GluR2, GluR3 and GluR4 and the AMPA receptors lacking GluR2 shows high Ca^<2+>-permeability. The roles of GluR2, of which the genetical symbol is GRIA2, in process formations in neurons and glial cells were examined using GRIA2-knock out mice, obtained form the Jackson Laboratory (Maine, USA). Although the knockout mice of GluR2 were introduced into Gunma University in 2004 and the propagation was made from the beginning of this project, the mice exhibited poor breeding rates. In heterozygote mice, GRIA2(+/-), One of two 11th exons of GRIA2 was substituted to a neomycin resistance gene by homologous recombination. Embryos of wild type mice, GRIA2(+/+), and knock-out mice, GRIA2(-/-) were obtained as follows. Females and males GRIA2(+/-) mice were mated overnight and separated the next morning and checked for the presence of gastration plug. This will be counted as gastration day 0.5(E0.5). Dissociated hippocampal culture was prepared from pups E16.5 or E17.5, killing the mother these embryos on the pregnancy 16.5 _17.5 days. For genotyping, PCR were performed according to the protocol of Jackson Laboratory Neurites of cultured neurons were thinner in GRIA2(-/-). The numbers of dendrites were fewer. In addition the spines of dendrites in glutamatergic neurons in GRIA2(-/-) mice were smaller than that in wild type. Since the limited periods of this grant, immunohistochemical analysis and electrophysiological experiments were not performed in the present study, remaining many subjects in the further study.

AMPA型谷氨酸受体由四个亚基组成，GLUR1，GLUR2，GLUR3和GLUR4，缺乏GlUR2的AMPA受体显示出很高的Ca^<2+> - 渗透性。 GLUR2的作用，其中遗传符号是Gria2，在神经元和神经胶质细胞中的过程形成中，使用Gria2敲除小鼠进行了研究，并形成了Jackson Laboratory（美国缅因州）的形式。尽管GLUR2的淘汰小鼠于2004年被引入Gunma University，并从该项目开始进行繁殖，但小鼠的育种率较差。在杂合子小鼠（+/-）的杂合小鼠中，通过同源重组将两个第11个外显子之一的Gria2外显子之一代替。获得野生型小鼠的胚胎，Gria2（+/+）和敲除小鼠，Gria2（ - / - ）的胚胎如下。雌性和男性Gria2（+/-）小鼠隔夜交配，并于第二天早晨分开，并检查是否存在胃塞。这将算作胃胃天0.5（E0.5）。通过幼犬E16.5或E17.5制备解离的海马培养，在妊娠16.5 _17.5天中杀死了母亲这些胚胎。对于基因分型，根据培养神经元的杰克逊实验室神经突的方案进行PCR（ - / - ）。树突的数量较少。此外，Gria2（ - / - ）小鼠中谷氨酸能神经元中树突的刺比野生型小鼠小。由于本研究中没有进行免疫组织化学分析和电生理实验，因此在本研究中未进行免疫组织化学分析和电生理实验，在进一步的研究中仍保留许多受试者。

项目成果

期刊论文数量（23）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Calcium dependence of aequorin bioluminescence dissected by random mutagenesis.

通过随机诱变剖析水母发光蛋白生物发光的钙依赖性。

DOI：
发表时间：
2006
期刊：
Proc Natl Acad Sci USA 103
影响因子：
0
作者：
Tricoire;Tsuzuki et al.
通讯作者：
Tsuzuki et al.

Membrane and firing properties of glutarnatergic and GABAergic neurons in the rat medial vestibular nucleus

大鼠内侧前庭核谷氨酸能和 GABA 能神经元的膜和放电特性

DOI：
发表时间：
2004
期刊：
J Neurophysiol 92
影响因子：
0
作者：
Takazawa;N. et al.
通讯作者：
N. et al.

Thermostable mutants of the photoprotein aequorin obtained by in vitro evolution

DOI：
10.1074/jbc.m505303200
发表时间：
2005-10-07
期刊：
JOURNAL OF BIOLOGICAL CHEMISTRY
影响因子：
4.8
作者：
Tsuzuki, K;Tricoire, L;Lambolez, B
通讯作者：
Lambolez, B

Identification and characterization of a novel Delphilin variant with an alternative N-terminus

具有替代 N 末端的新型 Delphilin 变体的鉴定和表征

DOI：
发表时间：
2005
期刊：
Mol. Brain Res. 141
影响因子：
0
作者：
Yamashita;T.;Miyagi;Y.;Ono;M.;Ito;H.;Watanabe;K.;Sonoda;T.;Tsuzuki;K.;Ozawa;S.;Aoki;I.;Okuda;K.;Mishina;M.;Kawamoto;S.
通讯作者：
S.

Serum-dependence of AMPA receptor-mediated proliferation in glioma cells