Glycine receptors (GlyRa1) - "Modulators for glycine receptors investigated using the Patchliner "
Patchliner application note: (0.4 MB)
Cells were kindly provided by AstraZeneca.
The Patchliner was used to study the pharmacology of the hGlyR a1 receptor expressed in mouse fibroblast cell line (L-tk). The Patchliner allows for fast application of drugs (<50 ms) with precisely controlled application intervals and wash times. These are important experimental parameters when investigating ligand gated ion channels and their effectors since many ligand gated ion channels rapidly desensitize. Kinetics and level of desensitization are determined by ligand concentration, exposure time, or both. Because of the Patchliner's rapid solution exchange combined with brief drug exposure, capabilities and timed application intervals, the deleterious effect caused by receptor desensitization can be minimized and corrected for. Here we show the use of Nanion's Patchliner for long (22 s) and short (1 s) application of glycine to patch clamped whole-cells. For the short applications of compound, a stacked solution application protocol was used. Specifically, two zones of solution were aspirated into the pipette used for administration. When applied to the cell, this results in exposure first to the agonist zone followed by a rapid wash out after 1 s of drug application. In all the protocols used, pre-incubation of modulators is possible, as well as washout in the presence of a modulator.