Determination of intracellular calcium in vivo via fluorine-19 nuclear magnetic resonance spectroscopy.
Determination of intracellular calcium in vivo via fluorine-19 nuclear magnetic resonance spectroscopy.
复制标题
通过氟 19 核磁共振波谱测定体内细胞内钙。
DOI：
10.1152/ajpcell.1995.269.2.c318
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Ackerman,JJ
中科院分区：
文献类型：
--
作者：
Song,SK;Hotchkiss,RS;Neil,J;MorrisJr,PE;Hsu,CY;Ackerman,JJ
NMR活性Ca 2+指示剂5-氟-1，2-双（2-氨基苯氧基）乙烷-N，N，N '，N'-四乙酸（5 F-BAPTA）的氟-19-核磁共振（19 F-NMR）光谱检测是一种用于测量胞质游离Ca 2+浓度（[Ca 2 +]i）的方法，并且先前已用于测量分离细胞和灌注器官中的[Ca 2 +]i。本研究的目的是证明使用19 F-NMR和5 F-BAPTA在体内和原位测定[Ca 2 +]i的可行性。实验在雄性Sprague-Dawley大鼠上进行，其中表面线圈天线用于NMR询问。Ca 2+指示剂5 F-BAPTA以100 mg/ml细胞渗透性乙酰氧基甲酯（5 F-BAPTA-AM）二甲亚砜溶液的形式静脉内（肾脏、脾脏）或脑室内（脑）输注。大鼠耐受静脉输注，平均动脉血压无明显变化。在检查的所有组织（肾脏、脾脏和脑）中，[Ca 2 +]i约为200 nM。据我们所知，这些结果代表了第一次在体内和原位测定[Ca 2 +]i采用19 F-NMR。
Fluorine-19-nuclear magnetic resonance (19F-NMR) spectroscopic detection of the NMR-active Ca2+ indicator 5-fluoro-1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (5F-BAPTA) is one method for measuring cytosolic free Ca2+ concentration ([Ca2+]i) and has been used previously to measure [Ca2+]i in isolated cells and perfused organs. The aim of the present investigation was to demonstrate the feasibility of determining [Ca2+]i in vivo and in situ using 19F-NMR and 5F-BAPTA. Experiments were performed on male Sprague-Dawley rats with a surface-coil antenna employed for NMR interrogation. The Ca2+ indicator, 5F-BAPTA, was infused either intravenously (kidney, spleen) or intraventricularly (brain) as a 100 mg/ml solution of the cell-permeant acetoxymethyl ester (5F-BAPTA-AM) in dimethyl sulfoxide. Rats tolerated intravenous infusion without evident change in mean arterial blood pressure. In all tissues examined, kidney, spleen, and brain, [Ca2+]i was approximately 200 nM. To our knowledge, these results represent the first in vivo and in situ determinations of [Ca2+]i employing 19F-NMR.