Studies on the alteration of calcium ions in osteoblastic cell line using a fluorescence dye

荧光染料对成骨细胞系钙离子变化的研究

基本信息

批准号：
01571018
负责人：
TAKAHASHI Kojiro
金额：
$ 1.34万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01571018/
关键词：
osteoblastic cells MC3T3-E1 cells epidermal carcinoma A431 cells influx of calcium ions intracellular calcium ions fluorescence indicator fluorescence microscopical system ガン細胞カルシウムイオンホルモン成長因子蛍光色素蛍光顕微測光

项目摘要

In mouse osteoblastic clone MC3T3-E1 cells, influx and intracellular alteration of Ca^<2+>ions were investigated with the fluorescence-spectrophotometer and fluorrescence-microscopic measurement system using a fluorescence reagent for intracellular Ca^<2+>ions. One passing influx of Ca^<2+>ions responsive to epidermal growth factor (EGF) was observed in human epidermoid carcinoma cell line : A431, but not in the osteoblastic cell line : MC3T3-E1 with both the fluorescence measurements. However, the increase of intracellular concentration of free Ca^<2+>ions responsive to fetal calf serum (FCS) was detected in both cell lines. With the manner of one passage, futhermore, influx of Ca^<2+>ions responsive to EGF appeared in the osteblastic MC3T3-E1 cells after the FCS-treatment, suggesting that a slight influx of Ca^<2+>ions into the MC3T3-E1 cells is induced with the synergistic function of EGF and any factor (s) in FCS.These results indicate that a very silght signal transduing due to the EGF-EGF receptor system is functioning by the synergistic effect during the initial growth phase of MC3T3-E1 cells. On the other hand, we also found that neurotrophin-3 (NT-3 : one of nerve growth factors) and its receptor (TRKC) is expressed in the osteoblastic MC3T3-E1 cells and the influx of Ca^<2+>ions responsive to NT-3 occurs in the osteoblastic cells. This implies that the signal transdicing sytem due to the interaction between NT-3 and TRKC is acting during the initial growth of MC3T3-E1.Moreover, photomapping data on the alteration of intracellular Ca^<2+>concentration detected with a fluorescence-microscopic measurement system indicated that the intracellular Ca^<2+>accumulation in MC3T3-E1 cells is higher than that in A431 cells. This suggests that an osteoblast-specific mechanism of intracellular Ca^<2+> accumulation plays in the osteoblastic cell system with the different manner from other cells such as epidermal cells.

在小鼠成骨细胞克隆MC3T3-E1细胞中，使用荧光型细胞胶质试剂进行荧光 - 微观测量系统研究了Ca^<2+>离子的流入和细胞内改变。在人类表皮类癌细胞系中观察到了一种对表皮生长因子（EGF）有反应的Ca^<2+>离子的涌入：A431，但在成骨细胞细胞系：MC3T3-E1中却没有两种荧光测量值。然而，在两种细胞系中都检测到了对胎儿小腿血清（FCS）反应的自由Ca^<2+离子的细胞内浓度的增加。 FCS处理后，以一段段落的方式（futhermore）涌入了对EGF的Ca^<2+>离子的涌入在成骨细胞MC3T3-E1细胞中出现MC3T3-E1细胞是通过EGF的协同功能和FC中的任何因素诱导的。这些结果表明，由于EGF-EGF受体系统引起的非常silght信号的传递，在同义效应中在同义效应中起作用。 MC3T3-E1细胞。另一方面，我们还发现Neurotrophin-3（NT-3：神经生长因子之一）及其受体（TRKC）在成骨细胞MC3T3-E1细胞中表达到NT-3发生在成骨细胞中。这意味着由于NT-3和TRKC之间相互作用引起的信号转导系统在MC3T3-E1的初始生长过程中起作用。此外，有关细胞内Ca^<2+>浓度改变的光映射数据，该数据用荧光 - 显微镜显微镜检测到的浓度检测测量系统表明，MC3T3-E1细胞中细胞内Ca^<2+>的积累高于A431细胞中的细胞内Ca^<2+>。这表明细胞内Ca^<2+>积累的成骨细胞特异性机制在成骨细胞系统中发挥作用，其方式与其他细胞（如表皮细胞）不同。