Efficient expression of a cnidarian peptide-gated ion channel in mammalian cells

Channels, vol. 15, pp. 273–283

Abstract

Hydra Na+ channels (HyNaCs) are peptide-gated ion channels of the DEG/ENaC gene family that are directly activated by neuropeptides of the Hydra nervous system. They have previously been successfully characterized in Xenopus oocytes. To establish their expression in mammalian cells, we transiently expressed heteromeric HyNaC2/3/5 in human HEK 293 and monkey COS-7 cells. We found that the expression of HyNaC2/3/5 using native cDNAs was inefficient and that codon optimization strongly increased protein expression and current amplitude in patch-clamp experiments. We used the improved expression of codon-optimized channel subunits to perform Ca2+ imaging and to demonstrate their glycosylation pattern. In summary, we established efficient expression of a cnidarian ion channel in mammalian cell lines.

Authors 4

1. Michèle Bachmann corresponding Aachen Department of Physiology

   RWTH Aachen University

   Affiliation as printed

   Department of Physiology, RWTH Aachen University, Aachen, Germany

2. Audrey Ortega-Ramírez Aachen Department of Physiology

   RWTH Aachen University

   Affiliation as printed

   Department of Physiology, RWTH Aachen University, Aachen, Germany

3. Lilia Leisle Aachen Department of Physiology

   RWTH Aachen University

   Affiliation as printed

   Department of Physiology, RWTH Aachen University, Aachen, Germany

4. Stefan Gründer Aachen Department of Physiology

   RWTH Aachen University

   Affiliation as printed

   Department of Physiology, RWTH Aachen University, Aachen, Germany