Application of photoactivated adenylyl cyclase to study the mechanisims of neurite formation by controlling temporal and spacial intracellular cAMP level in vitro

应用光激活腺苷酸环化酶体外控制细胞内时空cAMP水平研究神经突形成机制

• 批准号: 21590211
• 负责人: INOUE Akihiro
• 金额: $ 3万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21590211/
• 关键词: 神経突起形成; cAMP; 細胞内シグナル伝達; 光活性化; 光活性化アデニル酸シクラーゼ; エレクトロポレーション; メラノサイト; 神経突起伸展; アデニル酸シクラーゼ; PAC

Potencies of the photo-activated adenylyl cyclases(PAC) from euglena and bacteria for controlling intracellular cAMP concentration in vitro were analyzed biochemically with an cAMP-ELISA as well as cell-biologically with an Epac FRET sensor probe. It was also observed that melanosomes labeled with its associated protein Rab27a fused to fluorescent proteins were immediately moved toward peripheral regions of the B16 melanoma cells in response to an increase of the intracellular cAMP. We have also tried to utilize the PACs to study the morphological development and functions of neural cells in PC12 cells and hippocampal primary culture neurons.

从裸藻和细菌的光活化腺苷酸环化酶（PAC）在体外控制细胞内cAMP浓度的潜力进行了分析，生物化学与cAMP-ELISA以及细胞生物学与Epac FRET传感器探针。还观察到，用与荧光蛋白融合的相关蛋白Rab 27 a标记的黑素体响应于细胞内cAMP的增加而立即移向B16黑色素瘤细胞的外周区域。我们还尝试利用PAC对PC 12细胞和原代培养的海马神经元的形态发育和功能进行了研究。

项目成果

The role of kinase activity of Ca^<2+>/calmodulin-dependent protein kinase II in hippocampal synaptic plasticity and learning

Ca^2/钙调蛋白依赖性蛋白激酶II激酶活性在海马突触可塑性和学习中的作用

• 发表时间: 2009
• 作者: Yamagata Y.;et al.
• 通讯作者: et al.

Rewiring vesicle transports to probe the mechanism of polarized transports in neurons.神経細胞内の機能分子Trafficと高次脳機能そして疾患

重新连接囊泡运输以探究神经元中的极化运输机制、高级大脑功能和神经元内的疾病。

• 发表时间: 2010
• 作者: Takao Nakata;Nobutaka Hirokawa
• 通讯作者: Nobutaka Hirokawa

The kinesin-1 motor protein KIF5 recognizes microtubule lattice structure emanated from GTP-tubulin in axons as directional cue.(キネシンは、GTPチュブリンに由来する微小管格子構造を認識し、軸索の方向を認識する) Neuroscience 2011

驱动蛋白 1 运动蛋白 KIF5 识别轴突中 GTP 微管蛋白发出的微管晶格结构作为方向线索。

• 发表时间: 2011
• 作者: Takao Nakata;Shinsuke Niwa;Yasushi Okada;Franck Perez;Nobutaka Hirokawa
• 通讯作者: Nobutaka Hirokawa

Kinase-dead knock-in mouse reveals an essential role of kinase activity of Ca2+/calmodulin-dependent protein kinase IIalpha in dendritic spine enlargement, long-term potentiation, and learning.

激酶死亡敲入小鼠揭示了 Ca2/钙调蛋白依赖性蛋白激酶 IIα 的激酶活性在树突棘增大、长时程增强和学习中的重要作用。

• 发表时间: 2009
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Yamagata Y;Kobayashi S;Umeda T;Inoue A;Sakagami H;Fukaya M;Watanabe M;Hatanaka N;Totsuka M;Yagi T;Obata K;Imoto K;Yanagawa Y;Manabe T;Okabe S
• 通讯作者: Okabe S

ポリペプチド、単離された核酸、組み換えベクター、遺伝子導入キット、形質転換体、および細胞内カルシウムシグナルの調節方法

用于调节细胞内钙信号的多肽、分离的核酸、重组载体、基因转移试剂盒、转化体和方法

• 发表时间: 2012