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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GluR 1、GluR 2、GluR 3和GluR 4四个亚基组成，缺乏GluR 2的AMPA受体表现出高的Ca^2+渗透性。GluR 2（其遗传符号为GRIA 2）在神经元和神经胶质细胞中的突起形成中的作用使用GRIA 2敲除小鼠进行检查，所述GRIA 2敲除小鼠获自杰克逊实验室（缅因州，美国）。虽然2004年群马大学引进了GluR 2基因敲除小鼠，并从本项目开始就进行了繁殖，但小鼠的繁殖率很低。在杂合子小鼠中，GRIA 2（+/-），GRIA 2的两个第11外显子之一通过同源重组被替换为新霉素抗性基因。如下获得野生型小鼠GRIA 2（+/+）和敲除小鼠GRIA 2（-/-）的胚胎。将雌性和雄性GRIA 2（+/-）小鼠交配过夜，并在第二天早上分开，检查是否存在胃塞。这将计为第0.5天（E0.5）。从E16.5或E17.5的幼仔制备分离的海马培养物，在妊娠16.5 - 17.5天杀死这些胚胎。对于基因分型，根据杰克逊实验室的方案进行PCR。培养的神经元的神经突在GRIA 2（-/-）中较薄。树突数目较少。此外，GRIA 2（-/-）小鼠海马神经元树突棘较野生型小。由于研究时间有限，本研究未进行免疫组化分析和电生理实验，许多受试者有待进一步研究。

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）

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.

Predominant expression of GluR2 among the AMPA receptor subunits in neuronal progenitor cells of the rat hippocampus.

大鼠海马神经元祖细胞 AMPA 受体亚基中 GluR2 的主要表达。