Author:
Date:
2020-09-20
[Abstract] Ca2+ is an essential signaling messenger in all eukariotic cells, playing a pivotal role in many cellular functions as cell growth control (differentiation, fertilization and apoptosis), secretion, gene expression, enzyme regulation, among many others. This basic premise includes trypanosomatids as Trypanosoma cruzi and various species of Leishmania, the causative agents of Chagas disease and leishmaniasis respectively, where intracellular Ca2+ concentration ([Ca2+]i) has been demonstrated to be finely regulated. Nevertheless [Ca2+]i has been difficult to measure because of its very low cytoplasmic concentration (typically around 50-100 nM), when compared to the large concentration in the outside milieu ...
[摘要] [摘要] Ca 2+ 是所有真核细胞中必不可少的信号信使,在许多细胞功能中起着关键作用,例如细胞生长控制(分化,受精和凋亡),分泌,基因表达,酶调控等。这个基本前提包括锥虫如锥虫克氏锥虫和各种物种利什曼原虫,分别南美锥虫病和利什曼病的病原体,其中细胞内Ca 2+ 浓度([钙2 + ] 我)已被证明被精细地调节。然而,[Ca 2 + ] i 与外界环境中的高浓度(血液中约2 mM)相比,由于其细胞质浓度非常低(通常约为50-100 nM )而难以测量。细胞内荧光Ca 2+ 敏感指示剂的开发对于实现该目标至关重要。成功的基础是乙酰氧甲基化衍生物前体的合成,该合成物使通常由许多亲水性羧基组成的荧光分子(其非常亲和的Ca 2+ 结合(因此非常亲水))容易穿过质膜。一旦进入细胞内部,非特异性酯酶会将疏水部分从荧光骨架结构中分离出来,释放出羧基,然后将其转化为分子的酸形式,该分子保留在细胞质中并恢复其在Ca中发荧光的能力。2+ 依赖方式。其中,的Fura-2是迄今为止最常用的,因为它是一个比例(两个不同的波长激发和一个发射)的Ca 2+ 与钙指示剂2+ 与内[Ca亲和力兼容2 + ] 我。该协议主要包括用Fura-2加载指数相寄生虫,并通过双波长荧光分光光度计记录[Ca 2 + ] i 中的变化。这种技术允许有价值的信息AB的获取出的[Ca 2 + ] 我Ç ...
|