Yao Li-jun, Wang Gang, Ou-Yang Kun-fu, Wei Chao-liang, Wang Xian-hua, Wang Shi-rong, Yao Wei, Huang Hong-ping, Luo Jian-hong, Wu Cai-hong, Liu Jie, Zhou Zhuan, Cheng He-ping
Department of Neurobiology, Zhejiang University School of Medicine, Hangzhou 310031, China.
Acta Pharmacol Sin. 2006 Jul;27(7):848-52. doi: 10.1111/j.1745-7254.2006.00402.x.
Ca2+ release from the endoplasmic reticulum (ER) is an integral component of neuronal Ca2+ signaling. The present study is to investigate properties of local Ca2+ release events in superior cervical ganglion (SCG) neurons.
Primary cultured SCG neurons were prepared from neonatal rats (P3-P7). Low concentration of caffeine was used to induce Ca2+ release from the ER Ca2+ store, and intracellular Ca2+ was recorded by high-resolution line scan confocal imaging and the Ca2+ indicator Fluo-4.
Two populations of local Ca2+ release events with distinct temporal characteristics were evoked by 1.5 mmol/L caffeine near the surface membrane in the soma and the neurites of SCG neurons. Brief events similar to classic Ca2+ sparks lasted a few hundreds of milliseconds, whereas long-lasting events displayed duration up to tens of seconds. Typical somatic and neurite sparks were of 0.3- and 0.52-fold increase in local Fluo-4 fluorescence, respectively. Typical Ca2+ glows were brighter (deltaF/F0 approximately 0.6), but were highly confined in space. The half maximum of full duration of neurite sparks was much longer than those in the soma (685 vs 381 ms).
Co-existence of Ca2+ sparks and Ca2+ glows in SCG neurons indicates distinctive local regulation of Ca2+ release kinetics. The local Ca2+ signals of variable, site-specific temporal length may bear important implications in encoding a 'memory' of the trigger signal.
内质网(ER)释放Ca2+是神经元Ca2+信号传导的一个重要组成部分。本研究旨在探究颈上神经节(SCG)神经元中局部Ca2+释放事件的特性。
从新生大鼠(P3 - P7)制备原代培养的SCG神经元。使用低浓度咖啡因诱导内质网Ca2+储存库释放Ca2+,并通过高分辨率线扫描共聚焦成像和Ca2+指示剂Fluo - 4记录细胞内Ca2+。
1.5 mmol/L咖啡因在SCG神经元的胞体和神经突表面膜附近诱发了具有不同时间特征的两类局部Ca2+释放事件。类似于经典Ca2+闪烁的短暂事件持续几百毫秒,而持久事件的持续时间长达数十秒。典型的胞体和神经突闪烁分别使局部Fluo - 4荧光增加0.3倍和0.52倍。典型的Ca2+辉光更亮(ΔF/F0约为0.6),但在空间上高度局限。神经突闪烁的半峰全宽比胞体中的长得多(685对381毫秒)。
SCG神经元中Ca2+闪烁和Ca2+辉光共存表明Ca2+释放动力学存在独特的局部调节。可变的、位点特异性时间长度的局部Ca2+信号可能对触发信号的“记忆”编码具有重要意义。