Reducing agents facilitate membrane patch seal integrity and longevity.

Autor: Das D; Dept. of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, Canada., Wong A; Dept. of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, Canada., Friedman TN; Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Canada., Kerr BJ; Dept. of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, Canada.; Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Canada.; Department of Anesthesiology and Pain Medicine, University of Alberta, Edmonton, Canada., Kurata HT; Dept. of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, Canada.; Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Canada., Lamothe SM; Dept. of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, Canada.
Jazyk: angličtina
Zdroj: Channels (Austin, Tex.) [Channels (Austin)] 2024 Dec; Vol. 18 (1), pp. 2297621. Date of Electronic Publication: 2023 Dec 28.
DOI: 10.1080/19336950.2023.2297621
Abstrakt: The patch clamp method is a widely applied electrophysiological technique used to understand ion channel activity and cellular excitation. The formation of a high resistance giga-ohm seal is required to obtain high-quality recordings but can be challenging due to variables including operator experience and cell preparation. Therefore, the identification of methods to promote the formation and longevity of giga-ohm seals may be beneficial. In this report, we describe our observation that the application of reducing agents (DTT and TCEP) to the external bath solution during whole-cell patch clamp recordings of heterologous cells (HEK and LM) and cultured primary cells (DRG neurons) enhanced the success of giga-ohm seal formation. Reducing agents also maintained the integrity of the seal for longer periods of time at strong hyperpolarizing voltages, whereas an oxidizing agent (H 2 O 2 ) appeared to have the opposite effect. In summary, we report a useful tool to improve the quality of patch clamp recordings that may be helpful in certain experimental contexts.
Databáze: MEDLINE
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