How is intracellular calcium ion in parathyroidal tumor cells increased?

甲状旁腺肿瘤细胞胞内钙离子是如何增加的？

基本信息

批准号：
63570139
负责人：
GOTO Kunihiko
金额：
$ 1.02万
依托单位：
TOHOKU 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：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63570139/
关键词：
stimulation-secretion coupling intracellular calcium ion fura 2 calcium transition transient time fluorescence microscopy band pass filter emission 副甲状腺細胞 Fura2 Indo1 落射型倒立顕微鏡微山環境カルシウム指示薬副甲状腺腫瘍細胞 Fu-a2-AM GH_3細胞

项目摘要

In stimulation-secretion coupling, a stimulation induces the increase of intracellular calcium ions, subsequently following that the secretory granules are secreted by the mechanism of reverse pinocytosis. None hormone controls parathyroid chief cells and then by the increase or decrease of extracellular calcium ions the secretion of parathyroid hormone(PTH) is determined. In the present study, the measurement. of intracellular calcium ion transition and the relationship between calcium transition and secretion of PTH are investigated.The human and calf parathyroid tumor tissues were minced, and treated with 0.05% collagenase, subsequently resulting in preparing free cells. The dispersed parathyroid cells were cultured in the Pulbecco's minimum essential medium containing 10% fetal calf serum on a cover glass in Petri dish. They were treated with the loading of 5 muM Fura-2AM for 2 hours. After washing, the cover glass with the cultured cell was seton constant-temperature chamber, and perfused by lianks balanced salt solution. By project-type fluorescence phase-contrast microscopy, one cell was projected on emission of 340nm and 380 nm. The emission irradiated by one cell runs through 510nm band pass filter, and was received in ultra- sensitive video-camera. The images of the cell emission on the video-camera was expressed on oscillograph, and was drew on paper by penrecorder. The intracellular calcium ions was gradually increased in number for 5 msec after stimulation by hypocalcium concentration of perfusion fluid and them rapidly reached the highest point within the first 10 msec. The calcium transient was gradually decreased to basal point at 40 msec.

在刺激分泌耦合中，刺激会诱导细胞内钙离子的增加，随后，分泌颗粒是通过反向皮细胞增多的机制分泌的。无激素控制甲状旁腺主要细胞，然后通过增加或减少细胞外钙离子的增加或减少甲状旁腺激素（PTH）的分泌。在本研究中，测量。研究了细胞内钙离子过渡以及钙转变与PTH分泌之间的关系。将人和小腿甲状旁腺肿瘤组织切碎，并用0.05％的胶原酶处理，随后导致准备自由细胞。将分散的甲状旁腺细胞培养在Pulbecco的最低必需培养基中，该培养基在培养皿中的盖玻片上含有10％的胎牛血清。用5 Mum Fura-2am的负载治疗2小时。洗涤后，带有培养细胞的盖玻片是seton恒温室，并由lianks平衡盐溶液灌注。通过项目型荧光相对比显微镜，预测一个细胞在发射340nm和380 nm时。一个细胞辐照的发射通过510nm带通滤波器运行，并在超敏感的视频相机中接收。视频相机上的细胞发射的图像在示波器上表达，并由Penrecorder在纸上画。在灌注液的浓度下，细胞内钙离子的数量逐渐增加5毫秒，并在前10毫秒内迅速达到最高点。钙瞬态在40毫秒处逐渐降低至基础点。