Simultaneous measurements of cytosolic calcium and secretion in single bovine adrenal chromaffin cells by fluorescent imaging of fura-2 in cocultured cells.
Simultaneous measurements of cytosolic calcium and secretion in single bovine adrenal chromaffin cells by fluorescent imaging of fura-2 in cocultured cells.
复制标题
通过fura-2在共培养细胞中Fura-2的荧光成像，同时测量单牛肾上腺染色体细胞中的胞质钙和分泌。
DOI：
10.1083/jcb.109.3.1219
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发表时间：
1989-09
影响因子：
7.8
通讯作者：
Burgoyne, R D
中科院分区：
文献类型：
--
作者：
Cheek, T R;Jackson, T R;O'Sullivan, A J;Moreton, R B;Berridge, M J;Burgoyne, R D
胞浆游离钙浓度（[Ca 2 +]i）和胞吐的嗜铬颗粒同时从单一的，完整的牛肾上腺嗜铬细胞使用一种新的技术，涉及共培养细胞的荧光成像。用Fura-2监测嗜铬细胞[Ca 2 +]i。为了同时跟踪儿茶酚胺分泌，将细胞与负载Fura-2的NIH-3 T3 t细胞共培养，选择该细胞系是因为其对嗜铬细胞促分泌素无反应，但其对ATP的大Ca 2+反应，ATP与来自嗜铬细胞的儿茶酚胺共同释放。在去极化刺激尼古丁（一种有效的促分泌剂），嗜铬细胞[Ca 2 +]i迅速增加。在响应的峰值处，[Ca 2 +]i均匀地分布在整个细胞中。[Ca 2 +]i的这种升高随后是源自细胞整个表面的分泌反应。在响应肌醇1，4，5-三磷酸（InsP 3）动员激动剂血管紧张素II（弱促分泌素），观察到三种不同的反应。约30%的嗜铬细胞显示[Ca 2 +]i没有上升，不分泌。约45%的细胞响应于[Ca 2 +]i的大的（大于200 nM）瞬时升高，并且没有可检测的分泌响应。[Ca 2 +]i的升高是不均匀的，因此[Ca 2 +]i峰通常仅在细胞的一极记录。最后，大约25%的细胞产生了与上述类似的Ca 2+瞬时反应，但也产生了分泌反应。在这些情况下，分泌是极化的，被限制在细胞的极点，其中[Ca 2 +]i的上升是最大的。（250字处删节）
The cytosolic free calcium concentration ([Ca2+]i) and exocytosis of chromaffin granules were measured simultaneously from single, intact bovine adrenal chromaffin cells using a novel technique involving fluorescent imaging of cocultured cells. Chromaffin cell [Ca2+]i was monitored with fura-2. To simultaneously follow catecholamine secretion, the cells were cocultured with fura-2-loaded NIH-3T3t cells, a cell line chosen because of their irresponsiveness to chromaffin cell secretagogues but their large Ca2+ response to ATP, which is coreleased with catecholamine from the chromaffin cells. In response to the depolarizing stimulus nicotine (a potent secretagogue), chromaffin cell [Ca2+]i increased rapidly. At the peak of the response, [Ca2+]i was evenly distributed throughout the cell. This elevation in [Ca2+]i was followed by a secretory response which originated from the entire surface of the cell. In response to the inositol 1,4,5-trisphosphate (InsP3)-mobilizing agonist angiotensin II (a weak secretagogue), three different responses were observed. Approximately 30% of chromaffin cells showed no rise in [Ca2+]i and did not secrete. About 45% of the cells responded with a large (greater than 200 nM), transient elevation in [Ca2+]i and no detectable secretory response. The rise in [Ca2+]i was nonuniform, such that peak [Ca2+]i was often recorded only in one pole of the cell. And finally, approximately 25% of cells responded with a similar Ca2+-transient to that described above, but also gave a secretory response. In these cases secretion was polarized, being confined to the pole of the cell in which the rise in [Ca2+]i was greatest.(ABSTRACT TRUNCATED AT 250 WORDS)