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Aging and disease    2019, Vol. 10 Issue (3) : 626-636     DOI: 10.14336/AD.2018.0928
Review |
The WNK-SPAK/OSR1 Kinases and the Cation-Chloride Cotransporters as Therapeutic Targets for Neurological Diseases
Huachen Huang1,2, Shanshan Song2, Suneel Banerjee2, Tong Jiang2, Jinwei Zhang3, Kristopher T. Kahle4, Dandan Sun2,5,*, Zhongling Zhang1,*
1 Department of Neurology, The First Affiliate Hospital, Harbin Medical University, Harbin, Heilongjiang, China.
2Department of Neurology, University of Pittsburgh, Pittsburgh, PA, USA.
3Institute of Biomedical and Clinical Sciences, University of Exeter Medical School, Hatherly Laboratory, Exeter, EX4 4PS, UK.
4Departments of Neurosurgery, Pediatrics, and Cellular & Molecular Physiology, Centers for Mendelian Genomics, Yale School of Medicine, New Haven, CT, USA.
5Veterans Affairs Pittsburgh Health Care System, Geriatric Research, Education and Clinical Center, Pittsburgh, PA, USA.
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In recent years, cation-chloride cotransporters (CCCs) have drawn attention in the medical neuroscience research. CCCs include the family of Na+-coupled Cl- importers (NCC, NKCC1, and NKCC2), K+-coupled Cl- exporters (KCCs), and possibly polyamine transporters (CCC9) and CCC interacting protein (CIP1). For decades, CCCs have been the targets of several commonly used diuretic drugs, including hydrochlorothiazide, furosemide, and bumetanide. Genetic mutations of NCC and NKCC2 cause congenital renal tubular disorders and lead to renal salt-losing hypotension, secondary hyperreninemia, and hypokalemic metabolic alkalosis. New studies reveal that CCCs along with their regulatory WNK (Kinase with no lysine (K)), and SPAK (Ste20-related proline-alanine-rich kinase)/OSR1(oxidative stress-responsive kinase-1) are essential for regulating cell volume and maintaining ionic homeostasis in the nervous system, especially roles of the WNK-SPAK-NKCC1 signaling pathway in ischemic brain injury and hypersecretion of cerebrospinal fluid in post-hemorrhagic hydrocephalus. In addition, disruption of Cl- exporter KCC2 has an effect on synaptic inhibition, which may be involved in developing pain, epilepsy, and possibly some neuropsychiatric disorders. Interference with KCC3 leads to peripheral nervous system neuropathy as well as axon and nerve fiber swelling and psychosis. The WNK-SPAK/OSR1-CCCs complex emerges as therapeutic targets for multiple neurological diseases. This review will highlight these new findings.

Keywords brain edema      cell volume regulation      ischemic stroke      KCCs      NKCC1      WNK-SPAK/OSR1     
Corresponding Authors: Sun Dandan,Zhang Zhongling   
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These authors contributed equally to this work.

Just Accepted Date: 02 October 2018   Issue Date: 28 May 2019
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Huachen Huang
Shanshan Song
Suneel Banerjee
Tong Jiang
Jinwei Zhang
Kristopher T. Kahle
Dandan Sun
Zhongling Zhang
Cite this article:   
Huachen Huang,Shanshan Song,Suneel Banerjee, et al. The WNK-SPAK/OSR1 Kinases and the Cation-Chloride Cotransporters as Therapeutic Targets for Neurological Diseases[J]. Aging and disease, 2019, 10(3): 626-636.
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ProteinCo-transport ionsTissue and Subcellular DistributionRelating to human diseasesPhenotypes of KO/KI miceRefs.
SLC12A1 (NKCC2)Na+, K+, Cl-Kidney-specific (TAL);
apical plasma membrane; hypothalamo-neurohypophyseal system
Bartter syndromeSevere hypotension, hypokalemia,
hypercalciuria, metabolic alkalosis
SLC12A2 (NKCC1)Na+, K+, Cl-Ubiquitous;
basolateral plasma membrane
NoneSensorineural deafness,
alterations in endolymph secretion, reduced saliva production, sensory perception abnormalities and infertility
[58, 83-86]
SLC12A3 (NCC)Na+, Cl-Kidney-specific (DCT);
bone? apical plasma membrane
Hypotension, hypocalciuria, hypomagnesemia, hypokalemia[87-89]
SLC12A4 (KCC1)K+, Cl-UbiquitousNoneNo phenotype[90]
SLC12A5 (KCC2)K+, Cl-Neuron-specific;
basolateral plasma membrane (?)
EpilepsyComplete - death due to absent respiratory drive.
Incomplete KO (5%of function remains). Status epilepticus, death ± 15 days after birth.
[91, 92]
SLC12A6 (KCC3)K+, Cl-WidespreadAndermann syndromeKnockout mice recapitulate the
locomotion and neuropathy phenotypes and demonstrate axonal swelling
[92, 93]
SLC12A7 (KCC4)K+, Cl-Widespread;
basolateral plasma membrane (?)
NoneSensorineural deafness
and renal tubular acidosis
[94, 95]
SPAK--UbiquitousNo report in human, but resemble Gitelman syndrome in miceKO mice exhibited hypotension and recapitulated Gitelman syndrome with hypokalemia, hypomagnesemia, and hypocalciuria; higher nociceptive threshold and increased anxiety[12, 66]
OSR1--UbiquitousNo report in human, but resemble Bartter syndrome in miceGlobal KO - die in utero. Heterozygous KO - low BP. Kidney tubule-specific KO - normal BP with hypercalciuria and hypokalemia[96, 97]
Less susceptible to hypersensitivity to cold and mechanical stimuli after peripheral nerve injury[73, 75, 96, 98, 99]
WNK2--Prominently expressed in brain neurons; fetal brain and heartNo reportNo report[100, 101]
WNK3--UbiquitousAutistic disorder?Mice exhibited less cytotoxic edema after MCAO; compensated elevation of WNK1/SPAK axis in the kidney[34, 65, 72, 102]
WNK4--UbiquitousPHAIIKO mice exhibited hypokalemia but normalcalciuria[99, 103]
Table 1  Phenotypes of transgenic knockout (KO) mice involving WNK-SPAK/OSR1-CCC complex
Figure 1.  Roles of CCC in cell volume regulation

Intracellular osmolarity changes trigger cellular responses for volume regulation. Under hypertonic extracellular conditions, water extrudes from the cells and causes cell shrinkage, triggering a counter-response of regulatory volume increase (RVI). In this condition, the WNK-SPAK/OSR1 pathway is activated and phosphorylates NKCC1 and KCCs, resulting in NKCC1activation and KCCs inhibition. This subsequently leads to influx of Na+, K+ and Cl- via NKCC1 along with water, thus restoring cell volume. On the contrary, cell swelling due to hypotonic stress elicits regulatory volume decrease (RVD), in which the WNK-SPAK/OSR1 pathway remains inactive and NKCC1 and KCCs are dephosphorylated. This results in NKCC1 inhibition but stimulation of KCCs, which lead to KCC-mediated efflux of K+ and Cl- along with water, and cell volume decrease.

Figure 2.  Illustration of the WNK-SPAK/OSR1-CCCs cascade in nervous and non-nervous system diseases

Mutations of E3 ubiquitin ligase components cullin 3 (CUL3) and kelch-like 3 (KLHL3) were identified to cause Pseudohypoaldosteronism type II (PHAII) with increased WNK1 and WNK4 abundance in kidney. Gene mutations in WNK1 and WNK4 also cause PHAII with compromised cell volume homeostasis. In addition, osmotic stress can trigger WNK-SPAK/OSR1 complex activation, which leads to downstream phosphorylation of CCCs, especially stimulatory phosphorylation of NKCC1 and inhibitory phosphorylation of KCCs. Overstimulation of NKCC1 increases cytotoxic edema, enlarges infarction, and worsens neurobehavioral function in ischemic stroke. Hyperactive NKCC1 increases CSF secretion by the choroid plexus epithelium and causes post-hemorrhagic hydrocephalus after intraventricular hemorrhage. On the other hand, phosphorylation of KCC2 by WNK-SPAK/OSR1 decreases its Cl- efflux and reduces GABA-mediated inhibition of spinal nerve transmission and causes neuropathic pain. To date, no direct evidence links oxidative stress or inflammation to WNK activation in the nervous system. However, oxidative stress can directly activate SPAK/OSR1, which in turn regulates WNK activity, thus indirectly activates WNK; inflammation-induced stimulation of the WNK-SPAK/OSR1 pathway could also increase WNK activity. Dysfunction of KCC3, such as via KCC3 mutation, leads to compromised cell volume homeostasis and causes hereditary motor and sensory neuropathy with agenesis of the corpus callosum (HMSN/ACC), hearing loss and a reduced threshold for seizure. Hereditary sensory and autonomic neuropathy type II (HSANII) caused by HSN2 gene mutations leads to a loss-of-function for WNK1 activity. Taken together, the WNK-SPAK/OSR1-CCC signaling pathway emerges as a new therapeutic target for nervous and non-nervous system disorders.

[1] Arroyo JP, Kahle KT, Gamba G (2013). The SLC12 family of electroneutral cation-coupled chloride cotransporters. Mol Aspects Med, 34:288-298.
[2] Moriguchi T, Urushiyama S, Hisamoto N, Iemura S, Uchida S, Natsume T, et al. (2005). WNK1 regulates phosphorylation of cation-chloride-coupled cotransporters via the STE20-related kinases, SPAK and OSR1. J Biol Chem, 280:42685-42693.
[3] Vitari AC, Deak M, Morrice NA, Alessi DR (2005). The WNK1 and WNK4 protein kinases that are mutated in Gordon’s hypertension syndrome phosphorylate and activate SPAK and OSR1 protein kinases. Biochem J, 391:17-24.
[4] Richardson C, Alessi DR (2008). The regulation of salt transport and blood pressure by the WNK-SPAK/OSR1 signalling pathway. J Cell Sci, 121:3293-3304.
[5] Shekarabi M, Zhang J, Khanna AR, Ellison DH, Delpire E, Kahle KT (2017). WNK Kinase Signaling in Ion Homeostasis and Human Disease. Cell Metab, 25:285-299.
[6] Piala AT, Moon TM, Akella R, He H, Cobb MH, Goldsmith EJ (2014). Chloride sensing by WNK1 involves inhibition of autophosphorylation. Sci Signal, 7:ra41.
[7] Maruyama J, Kobayashi Y, Umeda T, Vandewalle A, Takeda K, Ichijo H, et al. (2016). Osmotic stress induces the phosphorylation of WNK4 Ser575 via the p38MAPK-MK pathway. Sci Rep, 6:18710.
[8] de Los Heros P, Alessi DR, Gourlay R, Campbell DG, Deak M, Macartney TJ, et al. (2014). The WNK-regulated SPAK/OSR1 kinases directly phosphorylate and inhibit the K+-Cl- co-transporters. Biochem J, 458:559-573.
[9] Choe KP, Strange K (2007). Evolutionarily conserved WNK and Ste20 kinases are essential for acute volume recovery and survival after hypertonic shrinkage in Caenorhabditis elegans. Am J Physiol Cell Physiol, 293:C915-927.
[10] Simon DB, Nelson-Williams C, Bia MJ, Ellison D, Karet FE, Molina AM, et al. (1996). Gitelman’s variant of Bartter’s syndrome, inherited hypokalaemic alkalosis, is caused by mutations in the thiazide-sensitive Na-Cl cotransporter. Nat Genet, 12:24-30.
[11] Naesens M, Steels P, Verberckmoes R, Vanrenterghem Y, Kuypers D (2004). Bartter’s and Gitelman’s syndromes: from gene to clinic. Nephron Physiol, 96:p65-78.
[12] Yang SS, Lo YF, Wu CC, Lin SW, Yeh CJ, Chu P, et al. (2010). SPAK-knockout mice manifest Gitelman syndrome and impaired vasoconstriction. J Am Soc Nephrol, 21:1868-1877.
[13] Seyberth HW, Schlingmann KP (2011). Bartter- and Gitelman-like syndromes: salt-losing tubulopathies with loop or DCT defects. Pediatr Nephrol, 26:1789-1802.
[14] Mercado A, Broumand V, Zandi-Nejad K, Enck AH, Mount DB (2006). A C-terminal domain in KCC2 confers constitutive K+-Cl- cotransport. J Biol Chem, 281:1016-1026.
[15] Hasbargen T, Ahmed MM, Miranpuri G, Li L, Kahle KT, Resnick D, et al. (2010). Role of NKCC1 and KCC2 in the development of chronic neuropathic pain following spinal cord injury. Ann N Y Acad Sci, 1198:168-172.
[16] Bhuiyan MIH, Song S, Yuan H, Begum G, Kofler J, Kahle KT, et al. (2017). WNK-Cab39-NKCC1 signaling increases the susceptibility to ischemic brain damage in hypertensive rats. J Cereb Blood Flow Metab, 37:2780-2794.
[17] Dzhala VI, Talos DM, Sdrulla DA, Brumback AC, Mathews GC, Benke TA, et al. (2005). NKCC1 transporter facilitates seizures in the developing brain. Nat Med, 11:1205-1213.
[18] Kaila K, Price TJ, Payne JA, Puskarjov M, Voipio J (2014). Cation-chloride cotransporters in neuronal development, plasticity and disease. Nat Rev Neurosci, 15:637-654.
[19] Kahle KT, Staley KJ, Nahed BV, Gamba G, Hebert SC, Lifton RP, et al. (2008). Roles of the cation-chloride cotransporters in neurological disease. Nat Clin Pract Neurol, 4:490-503.
[20] Pasantes-Morales H, Cardin V, Tuz K (2000). Signaling events during swelling and regulatory volume decrease. Neurochem Res, 25:1301-1314.
[21] McManus ML, Churchwell KB, Strange K (1995). Regulation of cell volume in health and disease. N Engl J Med, 333:1260-1266.
[22] Hoffmann EK, Lambert IH, Pedersen SF (2009). Physiology of cell volume regulation in vertebrates. Physiol Rev, 89:193-277.
[23] Pedersen SF, O’Donnell ME, Anderson SE, Cala PM (2006). Physiology and pathophysiology of Na+/H+ exchange and Na+ -K+ -2Cl- cotransport in the heart, brain, and blood. Am J Physiol Regul Integr Comp Physiol, 291:R1-25.
[24] Simard JM, Kent TA, Chen M, Tarasov KV, Gerzanich V (2007). Brain oedema in focal ischaemia: molecular pathophysiology and theoretical implications. Lancet Neurol, 6:258-268.
[25] McCormick JA, Ellison DH (2011). The WNKs: atypical protein kinases with pleiotropic actions. Physiol Rev, 91:177-219.
[26] Zhang J, Gao G, Begum G, Wang J, Khanna AR, Shmukler BE, et al. (2016). Functional kinomics establishes a critical node of volume-sensitive cation-Cl(-) cotransporter regulation in the mammalian brain. Sci Rep, 6:35986.
[27] Rinehart J, Maksimova YD, Tanis JE, Stone KL, Hodson CA, Zhang J, et al. (2009). Sites of regulated phosphorylation that control K-Cl cotransporter activity. Cell, 138:525-536.
[28] Adragna NC, Ravilla NB, Lauf PK, Begum G, Khanna AR, Sun D, et al. (2015). Regulated phosphorylation of the K-Cl cotransporter KCC3 is a molecular switch of intracellular potassium content and cell volume homeostasis. Front Cell Neurosci, 9:255.
[29] Jennings ML, Schulz RK (1991). Okadaic acid inhibition of KCl cotransport. Evidence that protein dephosphorylation is necessary for activation of transport by either cell swelling or N-ethylmaleimide. J Gen Physiol, 97:799-817.
[30] Thastrup JO, Rafiqi FH, Vitari AC, Pozo-Guisado E, Deak M, Mehellou Y, et al. (2012). SPAK/OSR1 regulate NKCC1 and WNK activity: analysis of WNK isoform interactions and activation by T-loop trans-autophosphorylation. Biochem J, 441:325-337.
[31] Markkanen M, Ludwig A, Khirug S, Pryazhnikov E, Soni S, Khiroug L, et al. (2017). Implications of the N-terminal heterogeneity for the neuronal K-Cl cotransporter KCC2 function. Brain Res, 1675:87-101.
[32] Vitari AC, Thastrup J, Rafiqi FH, Deak M, Morrice NA, Karlsson HK, et al. (2006). Functional interactions of the SPAK/OSR1 kinases with their upstream activator WNK1 and downstream substrate NKCC1. Biochem J, 397:223-231.
[33] Alessi DR, Zhang J, Khanna A, Hochdorfer T, Shang Y, Kahle KT (2014). The WNK-SPAK/OSR1 pathway: master regulator of cation-chloride cotransporters. Sci Signal, 7:re3.
[34] Zhao H, Nepomuceno R, Gao X, Foley LM, Wang S, Begum G, et al. (2017). Deletion of the WNK3-SPAK kinase complex in mice improves radiographic and clinical outcomes in malignant cerebral edema after ischemic stroke. J Cereb Blood Flow Metab, 37:550-563.
[35] Begum G, Yuan H, Kahle KT, Li L, Wang S, Shi Y, et al. (2015). Inhibition of WNK3 Kinase Signaling Reduces Brain Damage and Accelerates Neurological Recovery After Stroke. Stroke, 46:1956-1965.
[36] Pasantes-Morales H, Tuz K (2006). Volume changes in neurons: hyperexcitability and neuronal death. Contrib Nephrol, 152:221-240.
[37] Terker AS, Zhang C, Erspamer KJ, Gamba G, Yang CL, Ellison DH (2016). Unique chloride-sensing properties of WNK4 permit the distal nephron to modulate potassium homeostasis. Kidney Int, 89:127-134.
[38] Gagnon KB, England R, Delpire E (2006). Characterization of SPAK and OSR1, regulatory kinases of the Na-K-2Cl cotransporter. Mol Cell Biol, 26:689-698.
[39] Herbison AE, Moenter SM (2011). Depolarising and hyperpolarising actions of GABA(A) receptor activation on gonadotrophin-releasing hormone neurones: towards an emerging consensus. J Neuroendocrinol, 23:557-569.
[40] Kahle KT, Delpire E (2016). Kinase-KCC2 coupling: Cl- rheostasis, disease susceptibility, therapeutic target. J Neurophysiol, 115:8-18.
[41] Ivakine EA, Acton BA, Mahadevan V, Ormond J, Tang M, Pressey JC, et al. (2013). Neto2 is a KCC2 interacting protein required for neuronal Cl- regulation in hippocampal neurons. Proc Natl Acad Sci U S A, 110:3561-3566.
[42] Heubl M, Zhang J, Pressey JC, Al Awabdh S, Renner M, Gomez-Castro F, et al. (2017). GABAA receptor dependent synaptic inhibition rapidly tunes KCC2 activity via the Cl(-)-sensitive WNK1 kinase. Nat Commun, 8:1776.
[43] Ge S, Goh EL, Sailor KA, Kitabatake Y, Ming GL, Song H (2006). GABA regulates synaptic integration of newly generated neurons in the adult brain. Nature, 439:589-593.
[44] Spitzer NC (2010). How GABA generates depolarization. J Physiol, 588:757-758.
[45] Cellot G, Cherubini E (2014). GABAergic signaling as therapeutic target for autism spectrum disorders. Front Pediatr, 2:70.
[46] Arion D, Lewis DA (2011). Altered expression of regulators of the cortical chloride transporters NKCC1 and KCC2 in schizophrenia. Arch Gen Psychiatry, 68:21-31.
[47] Tornberg J, Voikar V, Savilahti H, Rauvala H, Airaksinen MS (2005). Behavioural phenotypes of hypomorphic KCC2-deficient mice. Eur J Neurosci, 21:1327-1337.
[48] Khanna A, Walcott BP, Kahle KT (2013). Limitations of Current GABA Agonists in Neonatal Seizures: Toward GABA Modulation Via the Targeting of Neuronal Cl(-) Transport. Front Neurol, 4:78.
[49] Boulenguez P, Liabeuf S, Bos R, Bras H, Jean-Xavier C, Brocard C, et al. (2010). Down-regulation of the potassium-chloride cotransporter KCC2 contributes to spasticity after spinal cord injury. Nat Med, 16:302-307.
[50] Rangroo Thrane V, Thrane AS, Wang F, Cotrina ML, Smith NA, Chen M, et al. (2013). Ammonia triggers neuronal disinhibition and seizures by impairing astrocyte potassium buffering. Nat Med, 19:1643-1648.
[51] Coull JA, Beggs S, Boudreau D, Boivin D, Tsuda M, Inoue K, et al. (2005). BDNF from microglia causes the shift in neuronal anion gradient underlying neuropathic pain. Nature, 438:1017-1021.
[52] Kahle KT, Khanna A, Clapham DE, Woolf CJ (2014). Therapeutic restoration of spinal inhibition via druggable enhancement of potassium-chloride cotransporter KCC2-mediated chloride extrusion in peripheral neuropathic pain. JAMA Neurol, 71:640-645.
[53] Sanchez-Brualla I, Boulenguez P, Brocard C, Liabeuf S, Viallat-Lieutaud A, Navarro X, et al. (2018). Activation of 5-HT2A Receptors Restores KCC2 Function and Reduces Neuropathic Pain after Spinal Cord Injury. Neuroscience, 387:48-57.
[54] Alvarez-Leefmans FJ, Gamino SM, Giraldez F, Nogueron I (1988). Intracellular chloride regulation in amphibian dorsal root ganglion neurones studied with ion-selective microelectrodes. J Physiol, 406:225-246.
[55] Kahle KT, Flores B, Bharucha-Goebel D, Zhang J, Donkervoort S, Hegde M, et al. (2016). Peripheral motor neuropathy is associated with defective kinase regulation of the KCC3 cotransporter. Sci Signal, 9:ra77.
[56] Andrew RD, Labron MW, Boehnke SE, Carnduff L, Kirov SA (2007). Physiological evidence that pyramidal neurons lack functional water channels. Cereb Cortex, 17:787-802.
[57] Payne JA, Stevenson TJ, Donaldson LF (1996). Molecular characterization of a putative K-Cl cotransporter in rat brain. A neuronal-specific isoform. J Biol Chem, 271:16245-16252.
[58] Dixon MJ, Gazzard J, Chaudhry SS, Sampson N, Schulte BA, Steel KP (1999). Mutation of the Na-K-Cl co-transporter gene Slc12a2 results in deafness in mice. Hum Mol Genet, 8:1579-1584.
[59] Adragna NC, Di Fulvio M, Lauf PK (2004). Regulation of K-Cl cotransport: from function to genes. J Membr Biol, 201:109-137.
[60] Howard HC, Mount DB, Rochefort D, Byun N, Dupre N, Lu J, et al. (2002). The K-Cl cotransporter KCC3 is mutant in a severe peripheral neuropathy associated with agenesis of the corpus callosum. Nat Genet, 32:384-392.
[61] Boettger T, Rust MB, Maier H, Seidenbecher T, Schweizer M, Keating DJ, et al. (2003). Loss of K-Cl co-transporter KCC3 causes deafness, neurodegeneration and reduced seizure threshold. Embo j, 22:5422-5434.
[62] Jiao Y, Jin X, Yan J, Zhang C, Jiao F, Li X, et al. (2008). A deletion mutation in Slc12a6 is associated with neuromuscular disease in gaxp mice. Genomics, 91:407-414.
[63] Byun N, Delpire E (2007). Axonal and periaxonal swelling precede peripheral neurodegeneration in KCC3 knockout mice. Neurobiol Dis, 28:39-51.
[64] Sun YT, Lin TS, Tzeng SF, Delpire E, Shen MR (2010). Deficiency of electroneutral K+-Cl- cotransporter 3 causes a disruption in impulse propagation along peripheral nerves. Glia, 58:1544-1552.
[65] Oi K, Sohara E, Rai T, Misawa M, Chiga M, Alessi DR, et al. (2012). A minor role of WNK3 in regulating phosphorylation of renal NKCC2 and NCC co-transporters in vivo. Biol Open, 1:120-127.
[66] Geng Y, Byun N, Delpire E (2010). Behavioral analysis of Ste20 kinase SPAK knockout mice. Behav Brain Res, 208:377-382.
[67] Brillault J, Lam TI, Rutkowsky JM, Foroutan S, O’Donnell ME (2008). Hypoxia effects on cell volume and ion uptake of cerebral microvascular endothelial cells. Am J Physiol Cell Physiol, 294:C88-96.
[68] Karimy JK, Zhang J, Kurland DB, Theriault BC, Duran D, Stokum JA, et al. (2017). Inflammation-dependent cerebrospinal fluid hypersecretion by the choroid plexus epithelium in posthemorrhagic hydrocephalus. Nat Med, 23:997-1003.
[69] Delpire E, Kahle KT (2017). The KCC3 cotransporter as a therapeutic target for peripheral neuropathy. Expert Opin Ther Targets, 21:113-116.
[70] Shekarabi M, Moldrich RX, Rasheed S, Salin-Cantegrel A, Laganiere J, Rochefort D, et al. (2012). Loss of neuronal potassium/chloride cotransporter 3 (KCC3) is responsible for the degenerative phenotype in a conditional mouse model of hereditary motor and sensory neuropathy associated with agenesis of the corpus callosum. J Neurosci, 32:3865-3876.
[71] Flores B, Schornak CC, Delpire E (2018). A role for KCC3 in maintaining cell volume of peripheral nerve fibers. Neurochem Int.
[72] Qiao Y, Liu X, Harvard C, Hildebrand MJ, Rajcan-Separovic E, Holden JJ, et al. (2008). Autism-associated familial microdeletion of Xp11.22. Clin Genet, 74:134-144.
[73] Lafreniere RG, MacDonald ML, Dube MP, MacFarlane J, O’Driscoll M, Brais B, et al. (2004). Identification of a novel gene (HSN2) causing hereditary sensory and autonomic neuropathy type II through the Study of Canadian Genetic Isolates. Am J Hum Genet, 74:1064-1073.
[74] Shekarabi M, Girard N, Riviere JB, Dion P, Houle M, Toulouse A, et al. (2008). Mutations in the nervous system--specific HSN2 exon of WNK1 cause hereditary sensory neuropathy type II. J Clin Invest, 118:2496-2505.
[75] Kahle KT, Schmouth JF, Lavastre V, Latremoliere A, Zhang J, Andrews N, et al. (2016). Inhibition of the kinase WNK1/HSN2 ameliorates neuropathic pain by restoring GABA inhibition. Sci Signal, 9:ra32.
[76] Hubner CA, Stein V, Hermans-Borgmeyer I, Meyer T, Ballanyi K, Jentsch TJ (2001). Disruption of KCC2 reveals an essential role of K-Cl cotransport already in early synaptic inhibition. Neuron, 30:515-524.
[77] Ohta A, Schumacher FR, Mehellou Y, Johnson C, Knebel A, Macartney TJ, et al. (2013). The CUL3-KLHL3 E3 ligase complex mutated in Gordon’s hypertension syndrome interacts with and ubiquitylates WNK isoforms: disease-causing mutations in KLHL3 and WNK4 disrupt interaction. Biochem J, 451:111-122.
[78] Shibata S, Zhang J, Puthumana J, Stone KL, Lifton RP (2013). Kelch-like 3 and Cullin 3 regulate electrolyte homeostasis via ubiquitination and degradation of WNK4. Proc Natl Acad Sci U S A, 110:7838-7843.
[79] Wakabayashi M, Mori T, Isobe K, Sohara E, Susa K, Araki Y, et al. (2013). Impaired KLHL3-mediated ubiquitination of WNK4 causes human hypertension. Cell Rep, 3:858-868.
[80] Adachi M, Asakura Y, Sato Y, Tajima T, Nakajima T, Yamamoto T, et al. (2007). Novel SLC12A1 (NKCC2) mutations in two families with Bartter syndrome type 1. Endocr J, 54:1003-1007.
[81] Konopacka A, Qiu J, Yao ST, Greenwood MP, Greenwood M, Lancaster T, et al. (2015). Osmoregulation requires brain expression of the renal Na-K-2Cl cotransporter NKCC2. J Neurosci, 35:5144-5155.
[82] Takahashi N, Chernavvsky DR, Gomez RA, Igarashi P, Gitelman HJ, Smithies O (2000). Uncompensated polyuria in a mouse model of Bartter’s syndrome. Proc Natl Acad Sci U S A, 97:5434-5439.
[83] Nezu A, Parvin MN, Turner RJ (2009). A conserved hydrophobic tetrad near the C terminus of the secretory Na+-K+-2Cl- cotransporter (NKCC1) is required for its correct intracellular processing. J Biol Chem, 284:6869-6876.
[84] Orlov SN, Koltsova SV, Kapilevich LV, Gusakova SV, Dulin NO (2015). NKCC1 and NKCC2: The pathogenetic role of cation-chloride cotransporters in hypertension. Genes Dis, 2:186-196.
[85] Nejsum LN, Praetorius J, Nielsen S (2005). NKCC1 and NHE1 are abundantly expressed in the basolateral plasma membrane of secretory coil cells in rat, mouse, and human sweat glands. Am J Physiol Cell Physiol, 289:C333-340.
[86] Delpire E, Lu J, England R, Dull C, Thorne T (1999). Deafness and imbalance associated with inactivation of the secretory Na-K-2Cl co-transporter. Nat Genet, 22:192-195.
[87] Nicolet-Barousse L, Blanchard A, Roux C, Pietri L, Bloch-Faure M, Kolta S, et al. (2005). Inactivation of the Na-Cl co-transporter (NCC) gene is associated with high BMD through both renal and bone mechanisms: analysis of patients with Gitelman syndrome and Ncc null mice. J Bone Miner Res, 20:799-808.
[88] Yang SS, Lo YF, Yu IS, Lin SW, Chang TH, Hsu YJ, et al. (2010). Generation and analysis of the thiazide-sensitive Na+ -Cl- cotransporter (Ncc/Slc12a3) Ser707X knockin mouse as a model of Gitelman syndrome. Hum Mutat, 31:1304-1315.
[89] Schultheis PJ, Lorenz JN, Meneton P, Nieman ML, Riddle TM, Flagella M, et al. (1998). Phenotype resembling Gitelman’s syndrome in mice lacking the apical Na+-Cl- cotransporter of the distal convoluted tubule. J Biol Chem, 273:29150-29155.
[90] Rust MB, Alper SL, Rudhard Y, Shmukler BE, Vicente R, Brugnara C, et al. (2007). Disruption of erythroid K-Cl cotransporters alters erythrocyte volume and partially rescues erythrocyte dehydration in SAD mice. J Clin Invest, 117:1708-1717.
[91] Blaesse P, Airaksinen MS, Rivera C, Kaila K (2009). Cation-chloride cotransporters and neuronal function. Neuron, 61:820-838.
[92] Delpire E, Mount DB (2002). Human and murine phenotypes associated with defects in cation-chloride cotransport. Annu Rev Physiol, 64:803-843.
[93] Rust MB, Faulhaber J, Budack MK, Pfeffer C, Maritzen T, Didie M, et al. (2006). Neurogenic mechanisms contribute to hypertension in mice with disruption of the K-Cl cotransporter KCC3. Circ Res, 98:549-556.
[94] Gamba G (2005). Molecular physiology and pathophysiology of electroneutral cation-chloride cotransporters. Physiol Rev, 85:423-493.
[95] Boettger T, Hubner CA, Maier H, Rust MB, Beck FX, Jentsch TJ (2002). Deafness and renal tubular acidosis in mice lacking the K-Cl co-transporter Kcc4. Nature, 416:874-878.
[96] Xie J, Yoon J, Yang SS, Lin SH, Huang CL (2013). WNK1 protein kinase regulates embryonic cardiovascular development through the OSR1 signaling cascade. J Biol Chem, 288:8566-8574.
[97] Lin SH, Yu IS, Jiang ST, Lin SW, Chu P, Chen A, et al. (2011). Impaired phosphorylation of Na(+)-K(+)-2Cl(-) cotransporter by oxidative stress-responsive kinase-1 deficiency manifests hypotension and Bartter-like syndrome. Proc Natl Acad Sci U S A, 108:17538-17543.
[98] Tu SW, Bugde A, Luby-Phelps K, Cobb MH (2011). WNK1 is required for mitosis and abscission. Proc Natl Acad Sci U S A, 108:1385-1390.
[99] Wilson FH, Disse-Nicodeme S, Choate KA, Ishikawa K, Nelson-Williams C, Desitter I, et al. (2001). Human hypertension caused by mutations in WNK kinases. Science, 293:1107-1112.
[100] Verissimo F, Jordan P (2001). WNK kinases, a novel protein kinase subfamily in multi-cellular organisms. Oncogene, 20:5562-5569.
[101] Hong C, Moorefield KS, Jun P, Aldape KD, Kharbanda S, Phillips HS, et al. (2007). Epigenome scans and cancer genome sequencing converge on WNK2, a kinase-independent suppressor of cell growth. Proc Natl Acad Sci U S A, 104:10974-10979.
[102] Mederle K, Mutig K, Paliege A, Carota I, Bachmann S, Castrop H, et al. (2013). Loss of WNK3 is compensated for by the WNK1/SPAK axis in the kidney of the mouse. Am J Physiol Renal Physiol, 304:F1198-1209.
[103] Terker AS, Castaneda-Bueno M, Ferdaus MZ, Cornelius RJ, Erspamer KJ, Su XT, et al. (2018). With no lysine kinase 4 modulates sodium potassium 2 chloride cotransporter activity in vivo. Am J Physiol Renal Physiol, 315:F781-f790.
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