Thomas Roger C, Postma Marten
Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3EG, UK.
Cell Calcium. 2007 Apr;41(4):365-78. doi: 10.1016/j.ceca.2006.07.010. Epub 2006 Sep 8.
We have used quartz Ca2+-sensitive microelectrodes (CASMs) in large voltage-clamped snail neurones to investigate the inward spread of Ca2+ after a brief depolarisation. Both steady state and [Ca2+]i transients changed with depth of penetration. When the CASM tip was within 20 microm of the far side of the cell the [Ca2+]i transient time to peak was 4.4+/-0.5s, rising to 14.7+/-0.7s at a distance of 80 microm. We estimate that the Ca2+ transients travelled centripetally at an average speed of 6 microm2 s(-1) and decreased in size by half over a distance of about 45 microm. Cyclopiazonic acid had little effect on the size and time to peak of Ca2+ transients but slowed their recovery significantly. This suggests that the endoplasmic reticulum curtails rather than reinforces the transients. Injecting the calcium buffer BAPTA made the Ca2+ transients more uniform in size and increased their times to peak and rates of recovery near the membrane. We have developed a computational model for the transients, which includes diffusion, uptake and Ca2+ extrusion. Good fits were obtained with a rather large apparent diffusion coefficient of about 90+/-20 microm2 s(-1). This may assist fast recovery by extrusion.
我们在大电压钳制的蜗牛神经元中使用了石英钙敏感微电极(CASMs),以研究短暂去极化后钙离子的内向扩散。稳态和[Ca2+]i瞬变均随穿透深度而变化。当CASM尖端位于细胞远侧20微米范围内时,[Ca2+]i瞬变达到峰值的时间为4.4±0.5秒,在80微米的距离处上升至14.7±0.7秒。我们估计钙离子瞬变以平均6微米2秒-1的速度向心传播,并且在约45微米的距离内大小减半。环匹阿尼酸对钙离子瞬变的大小和达到峰值的时间影响很小,但显著减慢了它们的恢复速度。这表明内质网抑制而非增强了瞬变。注入钙缓冲剂BAPTA使钙离子瞬变的大小更加均匀,并增加了它们在膜附近达到峰值的时间和恢复速率。我们已经开发了一个瞬变的计算模型,其中包括扩散、摄取和钙离子外排。使用约90±20微米2秒-1的相当大的表观扩散系数获得了良好的拟合。这可能有助于通过外排实现快速恢复。