Neuron specific calcium-binding Protein, calbrain, is involved in memory formation.

神经元特异性钙结合蛋白 calbrain 参与记忆形成。

• 批准号: 11694284
• 负责人: TOKUDA Masaaki
• 金额: $ 2.43万
• 依托单位: KAGAWA MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694284/
• 关键词: calbrain; CaM kinase II; LTP; S100; brain; hypothermia; NMDA; alcoholism; カルシウム結合蛋白質; 記憶; Ca^<2+>; calmodulin-dependent protein kinase II; calcineurin; NMR

A novel Ca2+-binding protein, termed calbrain, was isolated from a human brain cDNA library. The analysis of deduced amino acid sequence revealed that calbrain contains two putative EF-hand motifs that show significantly high homology to those of the calmodulin(CaM)family rather than two EF-hand protein families. Studies in vitro revealed that calbrain competitively inhibited CaM binding to Ca2+/calmodulin-dependent kinase II(Ki = 129 nM)and reduced its kinase activily and autophosphorylation.Ca2+/calmodulin-dependent protein kinase I(CaM-kinase I)in rat retina was shown to be present in rat retina. Developmental studies revealed that CaM-kinase I expression increased in accordance with postnatal development. Expression of CaM-kinase I in the retinas of rats raised in the complete darkness markedly decreased.CaMK-I was shown to mainly localize in the cytoplasm of the control and LTP-induced neurons, and a significant increase of immunoreactivity was observed in the latter neurons. A part of CaMK-I was found to translocate to the nuclei of LTP-induced hippocampal CA1 neurons.To investigate the mechanism of chronic cell death following postischemic hypothermia, the change of N-methyl-D-aspartate receptor(NMDAR)was examined in the CA1 subfield of the gerbil hippocampus. At 1 week following postischemic hypothermia(32 degrees Cx4 h), all CA1 neurons survived ; however, immunoreactivity of NMDAR1 increased in neuronal perikarya whereas decreased in dendrites in the CA1 neurons. LTP was also significantly depressed at 1 week after hypothermia.Morphological changes of the suprachiasmatic nucleus(SCN)of the hypothalamus were investigated in mice exhibiting intoxication signs of stages 2 or 3 after a short application term of 6% ethanol. Alterations in glial cells and neurons were examined using anti-glial fibrillary acidic protein(GFAP)and short-term ethanol exposure led to strong expression of GFAP-immunoreactivily(GFAP-IR)in the dorsomedial part of the SCN.

从人脑cDNA文库中分离到一种新的钙结合蛋白calbrain。推导的氨基酸序列的分析表明，calbrain包含两个推定的EF-手基序，显示出显着的高同源性的钙调素（CaM）家族，而不是两个EF-手蛋白家族。体外研究表明，钙脑竞争性抑制CaM与Ca 2 +/钙调素依赖性激酶II（Ki = 129 nM）的结合，降低其激酶活性和自磷酸化;大鼠视网膜中存在Ca 2 +/钙调素依赖性蛋白激酶I（CaM-kinase I）。发育研究表明，钙调蛋白激酶I的表达增加，按照出生后的发展。在完全黑暗中升高的大鼠视网膜中，CaM-I的表达明显降低，CaM-I主要定位于对照和LTP诱导的神经元的胞质中，后者的免疫反应性明显增强。为了探讨低温致海马CA 1区神经元慢性死亡的机制，本实验观察了沙土鼠海马CA 1区N-甲基-D-天冬氨酸受体（NMDAR）的变化。在1周后缺血低温（32度Cx4小时），所有的CA 1神经元存活，然而，NMDAR 1的免疫反应增加神经元胞体，而减少在CA 1神经元的树突。低温1周后LTP也明显降低。本实验观察了6%乙醇短期染毒后出现2、3期中毒症状的小鼠下丘脑视交叉上核（SCN）的形态学变化。胶质细胞和神经元的变化进行了检查，使用抗胶质细胞酸性蛋白（GFAP）和短期乙醇暴露导致强烈的表达GFAP-免疫反应（GFAP-IR）在SCN的背内侧部分。

项目成果

Yamashita K et al.: "Purification of bovine S100A12 from recombinant Escherichia coli."Protein Expr Purif. 16-1. 47-52 (1999)

Yamashita K 等人：“从重组大肠杆菌中纯化牛 S100A12。”Protein Expr Purif。

Satoriotomo I: "Short-term ethanol exposure alters calbindin D28k and glial fibrillary acidic protein immunoreactivity in hippocampus of mice."Brain Res. 879・1-2. 55-64 (2000)

Satoriomo I：“短期乙醇暴露改变小鼠海马中的钙结合蛋白 D28k 和神经胶质纤维酸性蛋白免疫反应性。”Brain Res. 879・1-2 (2000)。

Tsumura T: "The expression of Ca^<2+>calmodulin-dependent protein kinase I in rat retina is regulated by light stimulation."Vision Res. 39・19. 3165-3173 (1999)

Tsumura T：“大鼠视网膜中Ca ^ 2+ 钙调蛋白依赖性蛋白激酶I的表达受光刺激的调节。”Vision Res 39・19 (1999)。

Tsumura T: "The expression of Ca^<2+>/calmodulin-dependent protein kinase I in rat retina is regulatory light stimulation"Vision Res. 39・19. 3165-3173 (1999)

Tsumura T：“大鼠视网膜中Ca ^ 2+ /钙调蛋白依赖性蛋白激酶I的表达是调节性光刺激”Vision Res 39・19（1999）。

Tsumura T et al.: "The expression of Ca^<2+>/calmodulin-dependent protein kinase I in rat retina is regulated by light stimulation."Vision Res. 39-19. 3165-3173 (1999)

Tsumura T等人：“大鼠视网膜中Ca 2+ /钙调蛋白依赖性蛋白激酶I的表达受到光刺激的调节。”Vision Res。