Bright fluorescent chemosensor platforms for imaging endogenous pools of neuronal zinc
Bright fluorescent chemosensor platforms for imaging endogenous pools of neuronal zinc
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DOI：
10.1016/j.chembiol.2004.01.017
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发表时间：
2004-02-01
影响因子：
--
通讯作者：
Lippard, SJ
中科院分区：
文献类型：
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作者：
Chang, CJ;Nolan, EM;Lippard, SJ
制备了一系列基于荧光素平台的新型Zinpyr(ZP)荧光化学传感器，并对其用于神经元锌离子成像进行了评价。对同源ZP支架上的电负性取代模式进行了系统的综合考察，为通过控制无金属(APO)探针在生理pH下的荧光团pK(A)值和伴随的质子诱导干扰荧光来调节“OFF-ON”光诱导电子转移(PET)探针的荧光响应提供了基础。我们进一步确定了这些改进的光学工具在询问锌离子的金属神经化学方面的价值；新的ZP3荧光团成像了活体海马神经元和脑片中锌离子的内源性存储，包括首次在离体齿状回培养中检测到锌离子的荧光。我们的发现表明，仔细控制荧光基团PKA可以最大限度地减少质子诱导的载脂蛋白探针的荧光，而电负性取代为细胞研究提供了一种调整PET化学传感器的一般策略。除了为神经科学中的锌离子提供改进的光学工具外，这些结果还为创造用于生物应用的优质荧光探针提供了一个合理的起点。
A series of new fluorescent Zinpyr (ZP) chemosensors based on the fluorescein platform have been prepared and evaluated for imaging neuronal Zn2+. A systematic synthetic survey of electronegative substitution patterns on a homologous ZP scaffold provides a basis for tuning the fluorescence responses of "off-on" photoinduced electron transfer (PET) probes by controlling fluorophore pK(a) values and attendant proton-induced interfering fluorescence of the metal-free (apo) probes at physiological pH. We further establish the value of these improved optical tools for interrogating the metalloneurochemistry of Zn2+; the novel ZP3 fluorophore images endogenous stores of Zn2+ in live hippocampal neurons and slices, including the first fluorescence detection of Zn2+ in isolated dentate gyrus cultures. Our findings reveal that careful control of fluorophore pKa can minimize proton-induced fluorescence of the apo probes and that electronegative substitution offers a general strategy for tuning PET chemosensors for cellular studies. In addition to providing improved optical tools for Zn2+ in the neurosciences, these results afford a rational starting point for creating superior fluorescent probes for biological applications.