Department of Psychiatry, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
J Neurosci. 2010 Feb 24;30(8):2871-9. doi: 10.1523/JNEUROSCI.5607-09.2010.
Voltage-sensitive dyes are important tools for assessing network and single-cell excitability, but an untested premise in most cases is that the dyes do not interfere with the parameters (membrane potential, excitability) that they are designed to measure. We found that popular members of several different families of voltage-sensitive dyes modulate GABA(A) receptor with maximum efficacy and potency similar to clinically used GABA(A) receptor modulators. Di-4-ANEPPS and DiBAC4(3) potentiated GABA function with micromolar and high nanomolar potency, respectively, and yielded strong maximum effects similar to barbiturates and neurosteroids. Newer blue oxonols had biphasic effects on GABA(A) receptor function at nanomolar and micromolar concentrations, with maximum potentiation comparable to that of saturating benzodiazepine effects. ANNINE-6 and ANNINE-6plus had no detectable effect on GABA(A) receptor function. Even dyes with no activity on GABA(A) receptors at baseline induced photodynamic enhancement of GABA(A) receptors. The basal effects of dyes were sufficient to prolong IPSCs and to dampen network activity in multielectrode array recordings. Therefore, the dual effects of voltage-sensitive dyes on GABAergic inhibition require caution in dye use for studies of excitability and network activity.
电压敏感染料是评估网络和单细胞兴奋性的重要工具,但在大多数情况下,一个未经检验的前提是,这些染料不会干扰它们旨在测量的参数(膜电位、兴奋性)。我们发现,几种不同家族的电压敏感染料的常见成员以与临床使用的 GABA(A) 受体调节剂相似的最大功效和效力来调节 GABA(A) 受体。Di-4-ANEPPS 和 DiBAC4(3) 分别以微摩尔和高纳摩尔效力增强 GABA 的功能,产生类似于巴比妥类药物和神经甾体的强最大效应。新型蓝色氧杂蒽酮在纳摩尔和微摩尔浓度下对 GABA(A) 受体功能具有双相作用,最大增强作用可与饱和苯二氮䓬效应相媲美。ANNINE-6 和 ANNINE-6plus 对 GABA(A) 受体功能没有可检测到的影响。即使基线时对 GABA(A) 受体没有活性的染料也会诱导 GABA(A) 受体的光动力增强。染料的基础作用足以延长 IPSC 并抑制多电极阵列记录中的网络活动。因此,电压敏感染料对 GABA 能抑制的双重作用需要在研究兴奋性和网络活动时谨慎使用染料。