Bkaily G, Gros-Louis N, Naik R, Jaalouk D, Pothier P
Department of Physiology and Biophysics, Université de Sherbrooke, Quebec, Canada.
Mol Cell Biochem. 1996 Jan 26;154(2):113-21. doi: 10.1007/BF00226779.
In the present study, Fluo-3 Ca2+ measurement and confocal microscopy techniques were used in order to localize cytosolic []c and nuclear []n free Ca2+ distribution in resting and spontaneously contracting single heart cells from 10-day-old chick embryos. In resting single cells, the concentration of Ca2+ in the cytoplasm was lower than that in the nucleus. Increasing cytosolic free Ca2+ from 100-1600 nM gradually increased [Ca2+]n with a maximum capacity near 1200 nM. Results from Fura-2 microfluorometry and Fluo-3 confocal microscopy suggest a potential cross talk between the increase of cytosolic free Ca2+ and the uptake and release of Ca2+ by the nucleus during spontaneous contraction of single myocytes. Calcium waves in spontaneously contracting cells were found to spread from one cell to the next with the nucleus acting as a fluorescent beacon in which Ca2+ levels remained elevated for several milliseconds even after cytosolic Ca2+ had returned to near basal values. These results strongly suggest that the nucleus plays a negative and positive feedback role in controlling cytosolic free Ca2+ concentration during excitation-contraction coupling in heart cells.
在本研究中,采用Fluo-3钙离子测量和共聚焦显微镜技术,以定位10日龄鸡胚静止和自发收缩的单个心脏细胞中胞质[Ca²⁺]c和核[Ca²⁺]n的游离钙离子分布。在静止的单个细胞中,细胞质中钙离子的浓度低于细胞核中的浓度。将胞质游离钙离子浓度从100 - 1600 nM逐渐增加,会使[Ca²⁺]n逐渐增加,最大容量接近1200 nM。Fura-2显微荧光测定法和Fluo-3共聚焦显微镜的结果表明,在单个心肌细胞自发收缩过程中,胞质游离钙离子的增加与细胞核对钙离子的摄取和释放之间可能存在相互作用。发现自发收缩细胞中的钙波从一个细胞传播到下一个细胞,细胞核充当荧光信标,即使胞质钙离子已恢复到接近基础值,其中的钙离子水平仍会在几毫秒内保持升高。这些结果强烈表明,在心脏细胞兴奋 - 收缩偶联过程中,细胞核在控制胞质游离钙离子浓度方面发挥着负反馈和正反馈作用。