Potential interactions of calcium-sensitive reagents with zinc ion in different cultured cells.
Potential interactions of calcium-sensitive reagents with zinc ion in different cultured cells.
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DOI：
10.1371/journal.pone.0127421
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Yoneda Y
中科院分区：
文献类型：
--
作者：
Fujikawa K;Fukumori R;Nakamura S;Kutsukake T;Takarada T;Yoneda Y
几十年来，几种化学品已被广泛用于评估游离Ca 2+参与各种生物反应的机制。在这里，我们报告了高反应性锌的众所周知的Ca 2+敏感的试剂在不同的培养细胞。在大鼠星形胶质细胞C6胶质瘤细胞加载的荧光Ca 2+染料Fluo-3，另外的ZnCl 2逐渐增加的荧光强度的方式敏感的Ca 2+螯合剂EGTA，而不管添加的CaCl 2。Ca 2+离子载体A23187的添加在不存在ZnCl 2的情况下急剧增加Fluo-3荧光，而Zn 2+离子载体吡嗪的添加在ZnCl 2的存在下快速且另外增加荧光，但在其不存在下不增加荧光。在负载有锌染料FluoZin-3沿着Fluo-3的细胞中，在CaCl 2和ZnCl 2两者的存在下，Fluo-3的荧光类似地逐渐增加，但FluoZin-3的荧光没有增加。进一步加入吡啶并咪唑显著增加了两种染料的荧光强度，而加入Zn 2+螯合剂N，N，N ′，N ′-四（2-吡啶基甲基）乙烷-1，2-二胺（TPEN）则迅速并显著降低了FluoZin-3的荧光。在单独加载FluoZin-3的细胞中，ZnCl 2的添加以独立于添加的CaCl 2但对EGTA敏感的方式诱导荧光逐渐增加。ZnCl_2对C6胶质瘤细胞还原3-（4，5-二甲基-2-噻唑基）-2，5-二苯基-2H-溴化四氮唑的活性有明显的抑制作用，且吡啶酮可加速这种抑制作用。在多能P19细胞、神经元Neuro 2A细胞和小胶质细胞BV 2细胞中短暂暴露于ZnCl 2后，以EGTA敏感的方式发生了类似的抑制，这些细胞都表达特定锌转运蛋白的mRNA。两者合计，全面的分析是绝对需要的各种生理和病理反应的演示Ca 2+介导的不同细胞富含锌2+。
Several chemicals have been widely used to evaluate the involvement of free Ca2+ in mechanisms underlying a variety of biological responses for decades. Here, we report high reactivity to zinc of well-known Ca2+-sensitive reagents in diverse cultured cells. In rat astrocytic C6 glioma cells loaded with the fluorescent Ca2+ dye Fluo-3, the addition of ZnCl2 gradually increased the fluorescence intensity in a manner sensitive to the Ca2+ chelator EGTA irrespective of added CaCl2. The addition of the Ca2+ ionophore A23187 drastically increased Fluo-3 fluorescence in the absence of ZnCl2, while the addition of the Zn2+ ionophore pyrithione rapidly and additionally increased the fluorescence in the presence of ZnCl2, but not in its absence. In cells loaded with the zinc dye FluoZin-3 along with Fluo-3, a similarly gradual increase was seen in the fluorescence of Fluo-3, but not of FluoZin-3, in the presence of both CaCl2 and ZnCl2. Further addition of pyrithione drastically increased the fluorescence intensity of both dyes, while the addition of the Zn2+ chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethane-1,2-diamine (TPEN) rapidly and drastically decreased FluoZin-3 fluorescence. In cells loaded with FluoZin-3 alone, the addition of ZnCl2 induced a gradual increase in the fluorescence in a fashion independent of added CaCl2 but sensitive to EGTA. Significant inhibition was found in the vitality to reduce 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide in a manner sensitive to TPEN, EDTA and BAPTA in C6 glioma cells exposed to ZnCl2, with pyrithione accelerating the inhibition. Similar inhibition occurred in an EGTA-sensitive fashion after brief exposure to ZnCl2 in pluripotent P19 cells, neuronal Neuro2A cells and microglial BV2 cells, which all expressed mRNA for particular zinc transporters. Taken together, comprehensive analysis is absolutely required for the demonstration of a variety of physiological and pathological responses mediated by Ca2+ in diverse cells enriched of Zn2+.
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影响因子：
3.5
作者：
Fukumori, Ryo;Nakamichi, Noritaka;Yoneda, Yukio
通讯作者：
Yoneda, Yukio
影响因子：
5.3
作者：
Jing Qian;Noebels, Jeffrey L.
通讯作者：
Noebels, Jeffrey L.
影响因子：
3.3
作者：
Vander Jagt TA;Connor JA;Weiss JH;Shuttleworth CW
通讯作者：
Shuttleworth CW
影响因子：
16.2
作者：
Bossy-Wetzel, E;Talantova, MV;Lipton, SA
通讯作者：
Lipton, SA
影响因子：
56.9
作者：
BUSH, AI;PETTINGELL, WH;TANZI, RE
通讯作者：
TANZI, RE