Department of Physics, DIFILAB, University of Genoa, via Dodecaneso 33, 16146, Genoa, Italy.
Nanoscopy, CHT Erzelli, Istituto Italiano di Tecnologia, via E. Melen, 83, 16152, Genoa, Italy.
Biomol Concepts. 2022 Jun 8;13(1):289-297. doi: 10.1515/bmc-2022-0022.
The study of the GABA receptor itself and its pharmacology is of paramount importance for shedding light on the role of this receptor in the central nervous system. Caged compounds have emerged as powerful tools to support research in this field, as they allow to control, in space and time, the release of neurotransmitters enabling, for example, to map receptors' distribution and dynamics. Here we focus on γ-aminobutyric acid (GABA)-caged compounds, particularly on a commercial complex called RuBi-GABA, which has high efficiency of uncaging upon irradiation at visible wavelengths. We characterized, by electrophysiological measurements, the effects of RuBi-GABA on GABA receptors of rat cerebellar granule cells In particular, we evaluated the effects of side products obtained after RuBi-GABA photolysis. For this purpose, we developed a procedure to separate the "RuBi-cage" from GABA after uncaging RuBi-GABA with a laser source; then, we compared electrophysiological measurements acquired with and without administering the RuBi-cage in the perfusing bath. In conclusion, to investigate the role of the "cage" molecules both near and far from the cell soma, we compared experiments performed changing the distance of the uncaging point from the cell.
研究 GABA 受体本身及其药理学对于揭示该受体在中枢神经系统中的作用至关重要。笼状化合物已成为支持该领域研究的有力工具，因为它们可以控制神经递质的释放，在时间和空间上进行控制，从而能够例如，绘制受体的分布和动态图。在这里，我们重点介绍γ-氨基丁酸（GABA）笼状化合物，特别是一种名为 RuBi-GABA 的商业复合物，它在可见光波长照射下具有很高的解笼效率。我们通过电生理测量来表征 RuBi-GABA 对大鼠小脑颗粒细胞 GABA 受体的影响。特别是，我们评估了 RuBi-GABA 光解后获得的副产物的影响。为此，我们开发了一种在使用激光源解笼 RuBi-GABA 后从 GABA 中分离“RuBi-笼”的程序；然后，我们比较了在灌流浴中施用和不施用 RuBi-笼时获得的电生理测量结果。总之，为了研究“笼”分子在细胞体附近和远处的作用，我们比较了通过改变解笼点与细胞的距离来进行的实验。
