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首页> 外文期刊>Cell biology international. >EFFECTS OF K+-INDUCED DEPOLARIZATION AND PURINERGIC RECEPTOR ACTIVATION ON ELEMENTAL CONTENT IN INSULIN-PRODUCING RINM5F-CELLS
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EFFECTS OF K+-INDUCED DEPOLARIZATION AND PURINERGIC RECEPTOR ACTIVATION ON ELEMENTAL CONTENT IN INSULIN-PRODUCING RINM5F-CELLS

机译:K +诱导的去极化和肺受体活化对胰岛素产生的RINM5F细胞中元素含量的影响

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摘要

X-ray microanalysis was used to detect elemental changes in the insulin-producing tumor cell-line RINm5F. To improve discrimination between mobile ions and ions bound to macromolecules a new approach was employed, consisting of multivariate statistical analysis of correlations between the concentrations of Na, Mg, P, S, CI, K, and Ca. RINm5F cells, cultured an Formvar-coated titanium grids, were stimulated with high K+ or ATP, that are both known to stimulate insulin release. The buffers used contained Ca2+ or one of the Ca2+-analogues Sr2+ and Ba2+, to represent Ca2+ uptake in response to stimulation. After stimulation the cells were shock-frozen and freeze-dried overnight. Incubation for 10-20 seconds in a Ca2+-containing buffer did not significantly affect elemental composition, whereas cellular Mg, P and K decreased in a Sr2+-containing buffer. Depolarization with high K+ concentration caused an increase in the cellular Na content, both in Ca2+- and Sr2+-containing buffers, but not in the buffer where Ca2+ had been replaced by Ba2+. X-ray microanalysis is useful for detection of elemental changes subsequent to stimulation of cultured cells. Moreover, multivariate statistical analysis strengthens the idea that stimulation of RINm5F cells causes redistribution of ions possibly due to changes in the state of binding of some elements to cellular proteins.
机译:X射线微分析用于检测产生胰岛素的肿瘤细胞株RINm5F中的元素变化。为了改善移动离子与结合在大分子上的离子之间的区别,采用了一种新方法,该方法包括对Na,Mg,P,S,Cl,K和Ca浓度之间的相关性进行多变量统计分析。用高K +或ATP刺激培养有Formvar涂层的钛网格的RINm5F细胞,它们都刺激胰岛素释放。所使用的缓冲液包含Ca2 +或Ca2 +类似物Sr2 +和Ba2 +之一,以表示响应刺激而吸收Ca2 +。刺激后,将细胞骤冷并冷冻干燥过夜。在含Ca2 +的缓冲液中孵育10-20秒不会显着影响元素组成,而在含Sr2 +的缓冲液中细胞Mg,P和K降低。高K +浓度的去极化导致含Ca2 +和Sr2 +的缓冲液中细胞Na含量增加,但Ca2 +已被Ba2 +代替的缓冲液中却没有。 X射线微分析可用于检测刺激培养细胞后的元素变化。此外,多变量统计分析强化了这样一种观念,即RINm5F细胞的刺激可能导致离子的重新分布,这可能是由于某些元素与细胞蛋白结合状态的变化所致。

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