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Heterologous Expression of Aedes aegypti Cation Chloride Cotransporter 2 (aeCCC2) in Xenopus laevis Oocytes Induces an Enigmatic Na+/Li+ Conductance

1
Department of Entomology, The Ohio State University, Ohio Agricultural Research and Development Center, Wooster, OH 44691, USA
2
Department of Biology, Kenyon College, Gambier, OH 43022 USA
*
Author to whom correspondence should be addressed.
Insects 2019, 10(3), 71; https://doi.org/10.3390/insects10030071
Received: 17 February 2019 / Revised: 6 March 2019 / Accepted: 9 March 2019 / Published: 14 March 2019
(This article belongs to the Special Issue Integrative Mosquito Biology: From Molecules to Ecosystems)
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Abstract

The yellow fever mosquito Aedes aegypti possesses three genes encoding putative Na+-coupled cation chloride cotransporters (CCCs): aeNKCC1, aeCCC2, and aeCCC3. To date, none of the aeCCCs have been functionally characterized. Here we expressed aeCCC2 heterologously in Xenopus oocytes and measured the uptake of Li+ (a tracer for Na+) and Rb+ (a tracer for K+). Compared to control (H2O-injected) oocytes, the aeCCC2-expressing oocytes exhibited significantly greater uptake of Li+, but not Rb+. However, the uptake of Li+ was neither Cl-dependent nor inhibited by thiazide, loop diuretics, or amiloride, suggesting unconventional CCC activity. To determine if the Li+-uptake was mediated by a conductive pathway, we performed two-electrode voltage clamping (TEVC) on the oocytes. The aeCCC2 oocytes were characterized by an enhanced conductance for Li+ and Na+, but not K+, compared to control oocytes. It remains to be determined whether aeCCC2 directly mediates the Na+/Li+ conductance or whether heterologous expression of aeCCC2 stimulates an endogenous cation channel in the oocyte plasma membrane. View Full-Text
Keywords: Xenopus oocyte; mosquitoes; cation chloride cotransporters; two-electrode voltage clamping Xenopus oocyte; mosquitoes; cation chloride cotransporters; two-electrode voltage clamping
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Kalsi, M.; Gillen, C.; Piermarini, P.M. Heterologous Expression of Aedes aegypti Cation Chloride Cotransporter 2 (aeCCC2) in Xenopus laevis Oocytes Induces an Enigmatic Na+/Li+ Conductance. Insects 2019, 10, 71.

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