Electrophysiological study of the effects of side products of RuBi-GABA uncaging on GABAA receptors in cerebellar granule cells

Biomolecular Concepts

卷:13

Damonte Gianluca¹  Salis Annalisa¹  Robello Mauro²  Gatta Elena²  Diaspro Alberto²  Cupello Aroldo²  Angeli Elena²  Bazzurro Virginia² 

[1] Department of Experimental Medicine (DIMES), University of Genoa, via L.B. Alberti 2, 16132, Genoa, Italy;

[2] Department of Physics, DIFILAB, University of Genoa, via Dodecaneso 33, 16146, Genoa, Italy;

关键词: caged compound;    one-photon photoactivation;    whole-cell patch-clamp;    gabaa receptors;   

DOI  :  10.1515/bmc-2022-0022

来源: DOAJ

【 摘 要 】

The study of the GABAA 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 GABAA receptors of rat cerebellar granule cells in vitro. 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.

【 授权许可】

Unknown