Jenson, Lacey J; Sun, Baonan; Bloomquist, Jeffrey R
Voltage-sensitive potassium channels expressed after 20-Hydroxyecdysone treatment of a mosquito cell line
Teil von
  • Insect biochemistry and molecular biology, 2017-08-01, Vol.87, p.75-80
Ort / Verlag
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The goal of this research was to express receptors and ion channels in hormone-treated insect cell lines. Treatment of Anopheles gambiae Sua1B cells with 20-hydroxyecdysone showed an inhibition of cell growth over a time course of three days, with no change in cellular morphology. The effect of 20-hydroxyecdysone was enhanced in the presence of the potassium channel blocker 4-aminopyridine, but not tetraethylammonium. Concentration-response curves of 4-aminopyridine in the presence of 42 mu M (1 mg/ml) 20-hydroxyecdysone showed similar IC50 values (6-10 mu M) across 3 day exposures. Whole cell patch clamp confirmed the expression of delayed-rectifier (K-v2) potassium channels in hormone-supplemented Sua1B cells, whereas untreated Sua1B cells showed no evidence of Kv2 expression. The hormone-induced expression of K-v2 channels occurred in as little as 4 h after treatment, but were not observed after 24 h of exposure to 20-hydroxyecdysone, suggesting they played a role in cell death. The expressed channels had current-voltage relationships diagnostic for the K-v2 subtype, and were inhibited with an IC50 = 13 mM of tetraethylammonium. Overall, these parameters were similar to Anopheles gambiae Kv2 potassium channels expressed in HEK-293 cells. The induced presence of ion channels (and possibly receptors) in these cells has potential utility for high throughput screening and basic neuroscience research. (C) 2017 Elsevier Ltd. All rights reserved.

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