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Insect Biochem Mol Biol. 2017 Aug;87:75-80. doi: 10.1016/j.ibmb.2017.06.012. Epub 2017 Jun 28.

Voltage-sensitive potassium channels expressed after 20-Hydroxyecdysone treatment of a mosquito cell line.

Author information

1
Emerging Pathogens Institute, Department of Entomology and Nematology, University of Florida, Gainesville, FL, 32601, USA; Bedoukian Research Inc., 21 Finance Drive, Danbury, CT, 06810, USA.
2
Emerging Pathogens Institute, Department of Entomology and Nematology, University of Florida, Gainesville, FL, 32601, USA.
3
Emerging Pathogens Institute, Department of Entomology and Nematology, University of Florida, Gainesville, FL, 32601, USA. Electronic address: jbquist@epi.ufl.edu.

Abstract

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.

KEYWORDS:

20-Hydroxyecdysone; 4-Aminopyridine; Sua1B cells; Tetraethylammonium

PMID:
28668511
DOI:
10.1016/j.ibmb.2017.06.012
[Indexed for MEDLINE]

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