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Voltage-sensitive potassium channels expressed after 20-Hydroxyecdysone treatment of a mosquito cell line
Ist Teil von
Insect biochemistry and molecular biology, 2017-08, Vol.87, p.75-80
Ort / Verlag
England: Elsevier Ltd
Erscheinungsjahr
2017
Link zum Volltext
Quelle
Elsevier ScienceDirect Journals
Beschreibungen/Notizen
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 μM (1 mg/ml) 20-hydroxyecdysone showed similar IC50 values (6–10 μM) across 3 day exposures. Whole cell patch clamp confirmed the expression of delayed-rectifier (Kv2) potassium channels in hormone-supplemented Sua1B cells, whereas untreated Sua1B cells showed no evidence of Kv2 expression. The hormone-induced expression of Kv2 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 Kv2 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.
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•Sua1B cell growth was inhibited by 20-hydroxyecdysone.•This effect was enhanced by co-exposure to 4-aminopyridine.•K+ currents were expressed in hormone treated cells.•The effect was rapid, observed within 3 h, but not at 24 h.•The K+ currents had the expected properties.