{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T15:59:31Z","timestamp":1772812771806,"version":"3.50.1"},"reference-count":45,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2019,3,14]],"date-time":"2019-03-14T00:00:00Z","timestamp":1552521600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["NSF-IOS-1557230"],"award-info":[{"award-number":["NSF-IOS-1557230"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>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\u2212-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.<\/jats:p>","DOI":"10.3390\/insects10030071","type":"journal-article","created":{"date-parts":[[2019,3,15]],"date-time":"2019-03-15T04:12:09Z","timestamp":1552623129000},"page":"71","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Heterologous Expression of Aedes aegypti Cation Chloride Cotransporter 2 (aeCCC2) in Xenopus laevis Oocytes Induces an Enigmatic Na+\/Li+ Conductance"],"prefix":"10.3390","volume":"10","author":[{"given":"Megha","family":"Kalsi","sequence":"first","affiliation":[{"name":"Department of Entomology, The Ohio State University, Ohio Agricultural Research and Development Center, Wooster, OH 44691, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher","family":"Gillen","sequence":"additional","affiliation":[{"name":"Department of Biology, Kenyon College, Gambier, OH 43022 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1020-5774","authenticated-orcid":false,"given":"Peter M.","family":"Piermarini","sequence":"additional","affiliation":[{"name":"Department of Entomology, The Ohio State University, Ohio Agricultural Research and Development Center, Wooster, OH 44691, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,3,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1546","DOI":"10.1134\/S0006297914130070","article-title":"Cation-Chloride Cotransporters: Regulation, Physiological Significance, and Role in Pathogenesis of Arterial Hypertension","volume":"79","author":"Orlov","year":"2014","journal-title":"Biochem. Mosc."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1146\/annurev.physiol.64.081501.155847","article-title":"Human and murine phenotypes associated with defects in cation-chloride cotransport","volume":"64","author":"Delpire","year":"2002","journal-title":"Annu. Rev. Physiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1396","DOI":"10.1523\/JNEUROSCI.1421-05.2006","article-title":"The chloride transporter Na+-K+-Cl\u2212 cotransporter isoform-1 contributes to intracellular chloride increases after in vitro ischemia","volume":"26","author":"Pond","year":"2006","journal-title":"J. Neurosci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"16245","DOI":"10.1074\/jbc.271.27.16245","article-title":"Molecular characterization of a putative K-Cl cotransporter in rat brain\u2014A neuronal-specific isoform","volume":"271","author":"Payne","year":"1996","journal-title":"J. Biol. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2663","DOI":"10.1074\/jbc.M708194200","article-title":"Intramolecular and intermolecular fluorescence resonance energy transfer in fluorescent protein-tagged Na-K-Cl cotransporter (NKCC1)\u2014Sensitivity to regulatory conformational change and cell volume","volume":"283","author":"Pedersen","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"820","DOI":"10.1016\/j.neuron.2009.03.003","article-title":"Cation-Chloride Cotransporters and Neuronal Function","volume":"61","author":"Blaesse","year":"2009","journal-title":"Neuron"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1038\/nrn3819","article-title":"Cation-chloride cotransporters in neuronal development, plasticity and disease","volume":"15","author":"Kaila","year":"2014","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/B978-0-12-394316-3.00006-5","article-title":"Molecular Operation of the Cation Chloride Cotransporters: Ion Binding and Inhibitor Interaction","volume":"70","author":"Payne","year":"2012","journal-title":"Curr. Top. Membr."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"8943","DOI":"10.1523\/JNEUROSCI.4998-05.2006","article-title":"Mutations in the K+\/Cl\u2212 cotransporter gene kazachoc (kcc) increase seizure susceptibility in Drosophila","volume":"26","author":"Lundy","year":"2006","journal-title":"J. Neurosci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2599","DOI":"10.1242\/jeb.01058","article-title":"Basolateral ion transport mechanisms during fluid secretion by Drosophila Malpighian tubules: Na+ recycling, Na+: K+: 2Cl\u2212 cotransport and Cl\u2212 conductance","volume":"207","author":"Ianowski","year":"2004","journal-title":"J. Exp. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1111\/j.1748-1716.2010.02195.x","article-title":"Transcellular and paracellular pathways of transepithelial fluid secretion in Malpighian (renal) tubules of the yellow fever mosquito Aedes aegypti","volume":"202","author":"Beyenbach","year":"2011","journal-title":"Acta Physiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.pestbp.2018.03.019","article-title":"Use of chemical probes to explore the toxicological potential of the K+\/Cl\u2212 cotransporter (KCC) as a novel insecticide target to control the primary vector of dengue and Zika virus, Aedes aegypti","volume":"151","author":"Prael","year":"2018","journal-title":"Pestic Biochem. Phys."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"C883","DOI":"10.1152\/ajpcell.00201.2012","article-title":"The Drosophila NKCC Ncc69 is required for normal renal tubule function","volume":"303","author":"Rodan","year":"2012","journal-title":"Am. J. Physiol.-Cell Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1002\/glia.21103","article-title":"Drosophila Glia Use a Conserved Cotransporter Mechanism to Regulate Extracellular Volume","volume":"59","author":"Leiserson","year":"2011","journal-title":"Glia"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"C26","DOI":"10.1152\/ajpcell.00376.2009","article-title":"Developmental and functional studies of the SLC12 gene family members from Drosophila melanogaster","volume":"298","author":"Sun","year":"2010","journal-title":"Am. J. Physiol.-Cell Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.cbpa.2017.09.007","article-title":"Differential expression of putative sodium-dependent cation-chloride cotransporters in Aedes aegypti","volume":"214","author":"Piermarini","year":"2017","journal-title":"Comp. Biochem. Physiol. A Mol. Integr. Physiol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Durr, K.L., Tavraz, N.N., Spiller, S., and Friedrich, T. (2013). Measuring cation transport by Na,K- and H,K-ATPase in Xenopus oocytes by atomic absorption spectrophotometry: An alternative to radioisotope assays. J. Vis. Exp., e50201.","DOI":"10.3791\/50201-v"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"C437","DOI":"10.1152\/ajpcell.1996.270.2.C437","article-title":"Regulatory phosphorylation of the secretory Na-K-Cl cotransporter: Modulation by cytoplasmic Cl","volume":"270","author":"Lytle","year":"1996","journal-title":"Am. J. Physiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"37542","DOI":"10.1074\/jbc.M206293200","article-title":"A regulatory locus of phosphorylation in the N terminus of the Na-K-Cl cotransporter, NKCC1","volume":"277","author":"Darman","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"8458","DOI":"10.1073\/pnas.0802966105","article-title":"Regulation of NKCC2 by a chloride-sensing mechanism involving the WNK3 and SPAK kinases","volume":"105","author":"Kahle","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","first-page":"A183","article-title":"Osmotic Stimulation of Na+, K+, Cl\u2212 Cotransport in Squid Giant-Axon Is [Cl\u2212]I-Dependent","volume":"57","author":"Breitwieser","year":"1990","journal-title":"Biophys. J."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1016\/S0022-1910(00)00146-3","article-title":"Characterization of Rb uptake into Sf9 cells using cation chromatography: Evidence for a K-Cl cotransporter","volume":"47","author":"Bowles","year":"2001","journal-title":"J. Insect Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"F730","DOI":"10.1152\/ajprenal.90564.2008","article-title":"NHE8 is an intracellular cation\/H+ exchanger in renal tubules of the yellow fever mosquito Aedes aegypti","volume":"296","author":"Piermarini","year":"2009","journal-title":"Am. J. Physiol.-Renal."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of structural proteins during the assembly of the head of bacteriophage T4","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"R1318","DOI":"10.1152\/ajpregu.00223.2011","article-title":"Role of an apical K,Cl cotransporter in urine formation by renal tubules of the yellow fever mosquito (Aedes aegypti)","volume":"301","author":"Piermarini","year":"2011","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"39438","DOI":"10.1074\/jbc.M404671200","article-title":"Evidence for activation of endogenous transporters in Xenopus laevis oocytes expressing the Plasmodium falciparum chloroquine resistance transporter, PfCRT","volume":"279","author":"Nessler","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1113\/jphysiol.2008.161562","article-title":"NKCC1 (SLC12a2) induces a secondary axis in Xenopus laevis embryos independently of its co-transporter function","volume":"587","author":"Walters","year":"2009","journal-title":"J. Physiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1093\/cercor\/bhs027","article-title":"An Ion Transport-Independent Role for the Cation-Chloride Cotransporter KCC2 in Dendritic Spinogenesis In Vivo","volume":"23","author":"Fiumelli","year":"2013","journal-title":"Cereb Cortex"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2142","DOI":"10.1111\/j.1460-9568.2010.07258.x","article-title":"Premature expression of KCC2 in embryonic mice perturbs neural development by an ion transport-independent mechanism","volume":"31","author":"Horn","year":"2010","journal-title":"Eur. J. Neurosci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"G14","DOI":"10.1152\/ajpgi.00196.2003","article-title":"Ammonium effects on colonic Cl\u2212 secretion: Anomalous mole fraction behavior","volume":"286","author":"Worrell","year":"2004","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1007\/BF01869723","article-title":"Ammonium transport in medullary thick ascending limb of rabbit kidney: Involvement of the Na+,K+,Cl\u2212-cotransporter","volume":"94","author":"Kinne","year":"1986","journal-title":"J. Membr. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"344","DOI":"10.1038\/nrm2668","article-title":"Ion channels versus ion pumps: The principal difference, in principle","volume":"10","author":"Gadsby","year":"2009","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1007\/s11064-015-1699-6","article-title":"The Split Personality of Glutamate Transporters: A Chloride Channel and a Transporter","volume":"41","author":"Cater","year":"2016","journal-title":"Neurochem. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1038\/35014615","article-title":"An electroneutral sodium\/bicarbonate cotransporter NBCn1 and associated sodium channel","volume":"405","author":"Choi","year":"2000","journal-title":"Nature"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1080\/19336950.2018.1502585","article-title":"Cystic fibrosis transmembrane conductance regulator (CFTR): Making an ion channel out of an active transporter structure","volume":"12","author":"Linsdell","year":"2018","journal-title":"Channels"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/S0022-1910(02)00057-4","article-title":"Membrane conductances of principal cells in Malpighian tubules of Aedes aegypti","volume":"48","author":"Beyenbach","year":"2002","journal-title":"J. Insect Physiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"R339","DOI":"10.1152\/ajpregu.1985.248.3.R339","article-title":"Dibutyryl-cAMP increases basolateral sodium conductance of mosquito Malpighian tubules","volume":"248","author":"Sawyer","year":"1985","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"R379","DOI":"10.1152\/ajpregu.1985.249.4.R379","article-title":"Preliminary isolation of mosquito natriuretic factor","volume":"249","author":"Petzel","year":"1985","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3281","DOI":"10.1242\/jeb.01760","article-title":"Mosquito natriuretic peptide identified as a calcitonin-like diuretic hormone in Anopheles gambiae (Giles)","volume":"208","author":"Coast","year":"2005","journal-title":"J. Exp. Biol."},{"key":"ref_40","first-page":"R701","article-title":"Hormone-controlled cAMP-mediated fluid secretion in yellow-fever mosquito","volume":"253","author":"Petzel","year":"1987","journal-title":"Am. J. Physiol."},{"key":"ref_41","first-page":"171","article-title":"Diuresis in Mosquitoes Role of a Natriuretic Factor","volume":"2","author":"Beyenbach","year":"1987","journal-title":"News Physiol. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Kwon, H., Lu, H.L., Longnecker, M.T., and Pietrantonio, P.V. (2012). Role in diuresis of a calcitonin receptor (GPRCAL1) expressed in a distal-proximal gradient in renal organs of the mosquito Aedes aegypti (L.). PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0050374"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1007\/BF00690010","article-title":"Differential effects of secretagogues on Na and K secretion in the Malpighian tubules of Aedes aegypti","volume":"149","author":"Williams","year":"1983","journal-title":"J. Comp. Physiol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1007\/BF00689629","article-title":"Dynamic changes in flow rate and composition of urine during the post blood meal diuresis in Aedes aegypti","volume":"153","author":"Williams","year":"1983","journal-title":"J. Comp. Physiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1242\/jeb.024109","article-title":"Neuroendocrine control of ionic homeostasis in blood-sucking insects","volume":"212","author":"Coast","year":"2009","journal-title":"J. Exp. Biol."}],"container-title":["Insects"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4450\/10\/3\/71\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:38:51Z","timestamp":1760186331000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4450\/10\/3\/71"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,3,14]]},"references-count":45,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2019,3]]}},"alternative-id":["insects10030071"],"URL":"https:\/\/doi.org\/10.3390\/insects10030071","relation":{},"ISSN":["2075-4450"],"issn-type":[{"value":"2075-4450","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,3,14]]}}}