Automated fast perfusion of Xenopus oocytes for drug screening.
Fast ('concentration jump') applications of neurotransmitters are crucial for screening studies on ligand-gated ion channels. In this paper, we describe a method for automated fast perfusion of neurotransmitters (or drugs) during two-microelectrode voltage-clamp experiments on Xenopus oocytes. The oocytes are placed in a small bath chamber that is covered by a glass plate with two channels for the microelectrodes that are surrounded by a quartz funnel serving as a reservoir for test solutions. The oocytes are perfused in a vertical direction via the two channels in the plate. Automation of compound delivery is accomplished by means of a programmable pipetting workstation. A mean rise time for 10-90% current increase through muscle-type nACh channels of 55.0+/-1.3 ms (30 muM acetylcholine) was estimated. Automation, fast perfusion rates, and economical use of compounds ( approximately 100 mul/data point) make the system suitable for screening studies on ligand- and voltage-gated ion channels.
Baburin
I
I
Department of Pharmacology and Toxicology, University of Vienna, Althanstrasse 14, 1090 Vienna, Austria.
Beyl
S
S
Hering
S
S
D000818:Animals; D018929:Cell Culture Techniques; D015194:Diffusion Chambers, Culture; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation* / Q000379:methods; D008024:Ligands; D009865:Oocytes; D010477:Perfusion / Q000295:instrumentation* / Q000379:methods; D012371:Robotics; D013997:Time Factors; D014981:Xenopus
D008024:Ligands