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.

 抽象的

Hydra Na ⁺通道 (HyNaC) 是 DEG/ENaC 基因家族的肽门控离子通道，由Hydra神经系统的神经肽直接激活。此前已成功在非洲爪蟾卵母细胞中对它们进行了表征。为了确定它们在哺乳动物细胞中的表达，我们在人 HEK 293 和猴 COS-7 细胞中瞬时表达异聚 HyNaC2/3/5。我们发现使用天然 cDNA 表达 HyNaC2/3/5 效率低下，并且密码子优化在膜片钳实验中强烈增加了蛋白质表达和电流幅度。我们使用密码子优化的通道亚基的改进表达来进行 Ca ²⁺成像并证明其糖基化模式。总之，我们在哺乳动物细胞系中建立了刺胞动物离子通道的有效表达。