| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1989: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1988: ¥800,000 (Direct Cost: ¥800,000)
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| Research Abstract |
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.
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