Confocal microscopic imaging of [Ca2+]i in cultured rat hippocampal neurons following exposure to N‐methyl‐D‐aspartate.
Confocal microscopic imaging of [Ca2+]i in cultured rat hippocampal neurons following exposure to N‐methyl‐D‐aspartate.
复制标题
暴露于 N-甲基-D-天冬氨酸后培养的大鼠海马神经元中 [Ca2+]i 的共焦显微成像。
DOI：
--
复制
发表时间：
1992
期刊：
影响因子：
--
通讯作者：
D. Manor
中科院分区：
文献类型：
--
作者：
M. Segal;D. Manor
1.共聚焦激光扫描显微镜（CLSM）与钙指示剂染料Fluo-3联合使用，记录培养的海马神经元对N-甲基-D-天冬氨酸（NMDA）灌流的响应，细胞内游离钙浓度（[Ca 2 +]i）的变化。2. NMDA引起神经元各部位[Ca ~（2+）]i迅速升高。[Ca 2 +]i的升高依赖于NMDA受体的激活，通过去除Mg 2+和向灌流培养基中添加甘氨酸而增强，并且依赖于正常的[Ca 2 +]o。3. [Ca ~（2+）]i的升高首先见于近膜处。升高的[Ca 2 +]i波以117微米/秒的速率向心移动。4.丹曲林预孵育导致NMDA诱导的[Ca 2 +]i升高的有效性显著降低，表明至少部分升高是由细胞内钙释放引起的。5.钡替代钙引起的响应减少NMDA，但显着的反应仍然存在于这些细胞中，支持的假设，NMDA导致钙从细胞内存储的释放。6.从灌流培养基中去除钠延长了响应于NMDA的[Ca 2 +]i升高，表明Na-Ca反向转运蛋白有助于降低[Ca 2 +]i。7.这些研究证明了NMDA受体激活后[Ca 2 +]i调节机制的多样性。
1. The confocal laser scanning microscope (CLSM) was used in conjunction with the calcium indicator dye Fluo‐3 to record changes in free intracellular calcium concentration ([Ca2+]i) in cultured hippocampal neurons in response to superfusion of N‐methyl‐D‐aspartate (NMDA). 2. NMDA caused a rapid rise in [Ca2+]i in all parts of the neuron. The rise in [Ca2+]i was dependent on activation of an NMDA receptor, was enhanced by the removal of Mg2+ and addition of glycine to the superfusion medium, and was dependent on normal [Ca2+]o. 3. The rise of [Ca2+]i was seen first near the membrane. A wave of elevated [Ca2+]i moved centripetally at a rate of 117 microns/s. 4. Dantrolene pre‐incubation caused a significant reduction in the efficacy of the NMDA‐induced rise in [Ca2+]i, indicating that at least part of the rise is caused by intracellular release of calcium. 5. The replacement of calcium by barium caused a reduction in the response to NMDA, but a significant response was still present in these cells, supporting the assumption that NMDA causes release of calcium from intracellular stores. 6. The removal of sodium from the superfusion medium prolonged the [Ca2+]i rise in response to NMDA indicating that the Na‐Ca antiporter is instrumental in reducing [Ca2+]i. 7. These studies demonstrate the multiplicity of regulating mechanisms of [Ca2+]i following activation of NMDA receptors.