Affinage

Showing SLC12A2 — NKCC1 is a alias.

SLC12A2

Solute carrier family 12 member 2 · UniProt P55011

Length
1212 aa
Mass
131.4 kDa
Annotated
2026-06-10
100 papers in source corpus 40 papers cited in narrative 40 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 8/8 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

SLC12A2 (NKCC1) is an electroneutral Na+/K+/2Cl- cotransporter that couples ion movement to cell volume regulation and transepithelial salt transport across secretory, neural, and renal tissues (PMID:9556622). Kinetic reconstitution established its ion affinities and bumetanide sensitivity and showed that intracellular Cl- is the primary trigger of its activity under volume challenge (PMID:9556622). A series of cryo-EM structures of human and zebrafish NKCC1 defined a dimeric architecture with ion-binding sites formed at discontinuous TM1/TM6 helices, resolved inward-open, occluded, and outward-open conformations, and captured loop diuretics (bumetanide, furosemide) wedged into an extracellular translocation site plus a second cytosolic C-terminal site, delineating long-range conformational coupling between cytosolic and transmembrane domains (PMID:31367042, PMID:32081947, PMID:36239040, PMID:36306358). Transport is switched on by phosphorylation of N-terminal threonines (T203/T207/T212) by SPAK/OSR1 kinases acting downstream of WNK signaling; this phosphoregulatory domain interacts with the C-terminal domain to tune transport, as shown by both genetic and structural work (PMID:12740379, PMID:22032326, PMID:35585053, PMID:32413057). Membrane targeting requires N-glycosylation and is regulated post-translationally by Ca2+-dependent internalization, aldosterone-mediated protein stabilization, and Nedd4L-dependent (indirect) suppression of abundance (PMID:35585053, PMID:26351455, PMID:24173102, PMID:28087701). Functionally, NKCC1 localizes basolaterally in secretory epithelia and apically in choroid plexus, where phosphorylation-dependent activity drives CSF K+ clearance and protects against ventriculomegaly (PMID:15843440, PMID:9038823, PMID:33469018, PMID:36893755); it shapes neuronal Cl- homeostasis and network excitability (PMID:18394864), governs endolymph production required for hearing (PMID:10401008, PMID:19633174), suppresses renin secretion from juxtaglomerular cells (PMID:16106034), and acts cell-autonomously in microglia to restrain NLRP3 inflammasome activation (PMID:35085235). NKCC1 also couples cell-volume control to growth signaling, forming a complex with the LAT1-4F2hc leucine transporter to suppress mTORC1 (PMID:31067471). De novo loss-of-function mutations in SLC12A2 cause a neurodevelopmental disorder with bilateral sensorineural hearing loss (PMID:32658972).

Mechanistic history

Synthesis pass · year-by-year structured walk · 12 steps
  1. 1997 High

    Established where NKCC1 acts in the CNS and the secretory periphery, anchoring its physiological roles to specific membrane domains.

    Evidence In situ hybridization, immunocytochemistry, and 86Rb+ flux in polarized choroid plexus cultures

    PMID:9038823

    Open questions at the time
    • Did not resolve the molecular trafficking signals dictating apical versus basolateral targeting
    • Functional coupling to CSF homeostasis inferred, not directly perturbed
  2. 1998 High

    Defined NKCC1's transport stoichiometry and its regulation by intracellular Cl- and cell volume, distinguishing it kinetically from NKCC2.

    Evidence Stable HEK-293 expression with 86Rb+ uptake, ion substitution, and bumetanide kinetics

    PMID:9556622

    Open questions at the time
    • Molecular sensor of intracellular Cl- not identified
    • Did not connect Cl- sensing to a kinase pathway
  3. 1999 High

    Showed NKCC1 is genetically required for endolymph production and hearing, the first organismal loss-of-function phenotype.

    Evidence Positional cloning and cochlear analysis in shaker-with-syndactylism mouse mutants; confirmed in zebrafish otic vesicle

    PMID:10401008 PMID:19633174

    Open questions at the time
    • Cell-type responsible for endolymph defect within the cochlea not fully resolved
    • Did not address human disease relevance directly
  4. 2003 High

    Identified the activating kinase, showing SPAK phosphorylates N-terminal threonines and that a SPAK/phosphatase balance sets NKCC1 activity.

    Evidence Dominant-negative PASK/SPAK, 32Pi phosphorylation, Co-IP, and calyculin A rescue in HEK cells

    PMID:12740379

    Open questions at the time
    • Upstream activator of SPAK not yet placed
    • Did not map the precise threonines or structural consequence
  5. 2012 High

    Placed NKCC1 downstream of the WNK-SPAK/OSR1 axis genetically, proving SPAK/OSR1 activation is required for its phosphorylation.

    Evidence SPAK/OSR1 activation-deficient double-knockin ES cells with phospho-specific immunoblotting

    PMID:22032326

    Open questions at the time
    • Direct WNK-to-substrate kinetics not measured
    • Did not address tissue-specific contributions of SPAK versus OSR1
  6. 2019 High

    Provided the first structural framework for the CCC family, defining the ion-translocation pathway and domain communication of NKCC1.

    Evidence Cryo-EM of zebrafish NKCC1 with functional and MD validation

    PMID:31367042

    Open questions at the time
    • Single conformational snapshot; transport cycle not resolved
    • Phosphoregulatory domain not visualized
  7. 2022 High

    Resolved the human transport cycle and drug-binding mechanism across inward-open, occluded, ion-loaded, and outward-open/inhibitor-bound states, including the phosphoregulatory N-/C-domain interaction.

    Evidence Multiple single-particle cryo-EM structures of human NKCC1 with bumetanide/furosemide, ion-loaded states, and functional/MD validation

    PMID:32081947 PMID:33597714 PMID:35585053 PMID:36239040 PMID:36306358

    Open questions at the time
    • Phosphorylated versus dephosphorylated states not both captured at atomic detail
    • Dynamics of ion release along proposed TM5 pathway inferred from simulations
  8. 2020 High

    Connected the phosphoregulatory site to disease and to activity-state control via gain- and loss-of-function variants.

    Evidence Heterologous flux assays of p.Y199C (gain-of-function) and Xenopus oocyte assays of de novo loss-of-function variants with trio exome sequencing; patient fibroblast assays of a truncating variant

    PMID:26955005 PMID:27900370 PMID:32658972

    Open questions at the time
    • Genotype-phenotype relationship between gain- and loss-of-function variants not unified
    • In vivo neuronal consequences of variants not directly tested
  9. 2008 High

    Defined a developmental neuronal role: NKCC1 stabilizes network excitability rather than principally setting resting Cl-.

    Evidence NKCC1 KO mice, bumetanide pharmacology, Ca2+/Cl- imaging, and electrophysiology in CA3 neurons; complementary negative results in retina and hippocampal K+ clearance

    PMID:17493914 PMID:18394864 PMID:24482245

    Open questions at the time
    • Cell-type-specific Cl- contributions (neuron versus glia) remained partly unresolved
    • Did not address adult versus developmental switch mechanism
  10. 2019 High

    Linked NKCC1 cell-volume control to growth signaling through a physical complex with LAT1 that restrains mTORC1.

    Evidence Reciprocal Co-IP, NKCC1 KO/KD with mTORC1 assays in cells, colonic organoids, and mouse colon

    PMID:31067471

    Open questions at the time
    • Structural basis of the NKCC1-LAT1 interaction unknown
    • Whether transport activity per se or scaffolding drives mTORC1 suppression not fully separated
  11. 2023 High

    Established choroid plexus NKCC1 as a phosphorylation-dependent CSF K+ clearance pump that protects against hydrocephalus, and resolved its cell-type and splice-variant distribution in brain.

    Evidence ChP-targeted AAV-NKCC1 and phosphodeficient mutant in hydrocephalus and intraventricular blood models with CSF K+ and ventriculomegaly readouts; KO-validated IHC and single-cell analysis

    PMID:33469018 PMID:36573432 PMID:36893755

    Open questions at the time
    • Upstream Ca2+-to-phosphorylation coupling in ChP not fully mapped
    • Functional consequence of NKCC1a/NKCC1b splice variants not directly tested
  12. 2022 High

    Revealed a cell-autonomous immune role: microglial NKCC1 restrains NLRP3 inflammasome activation and limits ischemic injury.

    Evidence Microglia-specific conditional KO with morphology imaging, patch-clamp, NLRP3/IL-1β assays, and experimental stroke

    PMID:35085235

    Open questions at the time
    • Molecular link between NKCC1-driven volume change and NLRP3 priming not defined
    • Did not test relevance to human neuroinflammatory disease

Open questions

Synthesis pass · forward-looking unresolved questions
  • How phosphorylation-driven N-/C-domain reconfiguration is structurally coupled to the ion-transport cycle, and how the same transporter is partitioned among its diverse tissue-specific roles, remains to be unified.
  • No atomic structure comparing phospho- and dephospho-states
  • Mechanism selecting apical versus basolateral targeting unresolved
  • Functional division of labor between NKCC1a and NKCC1b splice variants untested

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0005215 transporter activity 4 GO:0140104 molecular carrier activity 2
Localization
GO:0005886 plasma membrane 4
Pathway
R-HSA-162582 Signal Transduction 4 R-HSA-382551 Transport of small molecules 2
Partners
LAT1/SLC7A5OSR1/OXSR1SLC3A2/4F2HCSPAK/STK39
Complex memberships
NKCC1-LAT1-4F2hc complex

Evidence

Reading pass · 40 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2003 PASK (SPAK) phosphorylates two N-terminal threonines of NKCC1 and is required for its activation. Dominant-negative PASK drastically reduces NKCC1 activity (60–80%), and this inhibition is rescued by the phosphatase inhibitor calyculin A, demonstrating that PASK/phosphatase balance controls NKCC1 phosphorylation state. Co-immunoprecipitation confirmed PASK–NKCC1 binding in HEK cells; the association is constitutive and not regulated by PASK or NKCC1 activity. Dominant-negative overexpression, 32Pi phosphorylation assay, co-immunoprecipitation, calyculin A rescue in HEK cells The Journal of biological chemistry High 12740379
2012 SPAK and OSR1 are essential intermediaries for WNK-dependent NKCC1 phosphorylation and activation. Double-knockin ES cells in which SPAK and OSR1 cannot be activated by WNK1 show complete loss of NKCC1 phosphorylation and activation, providing genetic proof that SPAK/OSR1 activity is required for NKCC1 function in the WNK pathway. Double-knockin ES cells (SPAK/OSR1 activation-deficient), immunoblotting with phospho-specific antibodies The Biochemical journal High 22032326
2019 Cryo-EM structure of zebrafish NKCC1 (Danio rerio) defined the overall architecture of the CCC family, identified the ion-translocation pathway, ion-binding sites, and key transmembrane residues required for transport activity, and revealed how cytosolic and transmembrane domains communicate for coupled ion transport. Cryo-electron microscopy, functional characterization, computational (MD) simulations Nature High 31367042
2020 Cryo-EM structure of human NKCC1 in a partially loaded, inward-open state revealed a dimeric assembly; TM1 and TM6 helices break α-helical geometry at ion-binding sites; multiple extracellular entryways and intracellular exits suggest K+, Na+, and Cl- may traverse distinct routes during translocation. Single-particle cryo-electron microscopy Nature communications High 32081947
2021 Cryo-EM structures of human NKCC1 and mouse KCC2 identified essential residues for ion transport and phosphorylation-dependent regulation, proposing a mechanism by which phosphorylation of the N-terminal regulatory domain modulates transport activity. Cryo-electron microscopy, computational analysis, functional characterization Communications biology High 33597714
2022 Cryo-EM structures of human NKCC1 in outward-facing conformation with bumetanide bound revealed the drug wedged into the extracellular ion translocation pathway. Structures also defined an N-terminal phosphoregulatory domain that interacts with the C-terminal domain, suggesting (de)phosphorylation regulates NKCC1 by tuning the strength of this intramolecular domain association. Single-particle cryo-EM, functional bumetanide-binding and transport assays Nature communications High 35585053
2022 2.6 Å cryo-EM structure of human NKCC1 in a substrate-loaded (Na+, K+, 2Cl-) occluded inward-facing state identified Cl- binding at the Cl1 site providing a structural bridge between scaffold and bundle domains, Cl- at Cl2 site undertaking a role analogous to a conserved glutamate in SLC6 transporters, and a putative Na+ release pathway along TM helix 5 coupled to the Cl2 site. Cryo-electron microscopy (2.6 Å), functional studies in mammalian cells, molecular dynamics simulations The EMBO journal High 36239040
2022 Cryo-EM structures of human NKCC1 in the absence and presence of loop diuretics (bumetanide or furosemide) revealed two drug-binding sites: one at the transmembrane domain and one at the cytosolic C-terminal domain. An inhibition mechanism involving coupled movement between cytosolic and transmembrane domains (long-range conformational coupling) was delineated. Single-particle cryo-EM (four structures), with and without bumetanide/furosemide Science advances High 36306358
1997 BSC2 (NKCC1) protein is localized to the apical surface of choroid plexus epithelium and to cell bodies/dendrites of neurons. Apical localization in choroid plexus was confirmed by 86Rb+ uptake in polarized primary cultures and confocal immunofluorescence, supporting a role in CSF K+ homeostasis. In situ hybridization, immunocytochemistry, 86Rb+ flux assay in polarized choroid plexus cell cultures, confocal microscopy The American journal of physiology High 9038823
1997 The Slc12a2 gene is encoded by 27 exons. An alternatively spliced variant lacking exon 21 (encoding a 16-amino-acid peptide in the C-terminal tail) is expressed primarily in brain; loss of this exon eliminates the single protein kinase A consensus site of the cotransporter, linking alternative splicing to differential regulation. RNase protection assay, primer extension, reporter gene transfection, nucleotide sequencing The American journal of physiology Medium 9357771
1998 Kinetic characterization of NKCC1 in HEK-293 cells established ion affinities (Na, K/Rb, Cl) and bumetanide affinity. NKCC1 activity is activated by low intracellular Cl- and responds to cell volume changes. Internal Cl- concentration is the primary driver of NKCC1 regulation under volume challenge, whereas NKCC2 responds preferentially to volume. Stable heterologous expression in HEK-293 cells, 86Rb+ uptake, ion substitution, bumetanide inhibition kinetics The Journal of biological chemistry High 9556622
1999 Loss-of-function mutations in Slc12a2 (Nkcc1) cause deafness in the shaker-with-syndactylism (sy and sy(ns)) mouse mutants, associated with abnormal endolymph production, establishing NKCC1 as a required component of K+ recycling in the cochlea. Positional candidate cloning, mutant allele identification, cochlear morphology analysis in knockout mice Human molecular genetics High 10401008
2009 In zebrafish, nkcc1 (slc12a2) loss-of-function mutations cause collapse of the otic vesicle (endolymph loss) and over-inflation of the swim bladder, with concomitant downregulation of genes involved in endolymph production, establishing NKCC1 as required for endolymph volume regulation in the inner ear. Genetic mapping, point mutation identification, morpholino splice-blocking, zebrafish larval phenotype analysis Development High 19633174
2005 NKCC1-deficient mice show elevated basal plasma renin concentration (~3-fold), and juxtaglomerular (JG) granular cells from NKCC1-null mice fail to increase membrane capacitance or renin release in response to furosemide, demonstrating that NKCC1 directly suppresses basal renin secretion from JG cells. NKCC1 knockout mice, plasma renin measurements, patch-clamp capacitance assay on single JG cells, primary JG cell cultures American journal of physiology. Renal physiology High 16106034
2008 Genetic deletion of NKCC1 in P9–P13 CA3 pyramidal neurons increases cell excitability and 4-aminopyridine-induced seizure-like activity. NKCC1 absence only marginally reduces resting intracellular Cl-, but large Cl- rises occur during network hyperexcitability (blocked by DNQX), indicating NKCC1's primary role at this stage is network stabilization rather than setting resting Cl-. NKCC1 knockout mice, bumetanide pharmacology, calcium imaging (fura-2), Cl- imaging (MQAE), electrophysiology Epilepsy research High 18394864
2012 Estradiol increases protein levels of SPAK and OSR1 in the neonatal rat hypothalamus via a transcription-dependent mechanism, and SPAK/OSR1 upregulation mediates estradiol-enhanced phosphorylation and activity of NKCC1. SPAK knockdown (and to a lesser degree OSR1 knockdown) abolishes estradiol-enhanced NKCC1 phosphorylation and GABA-induced Ca2+ influx. Antisense oligonucleotide knockdown of SPAK/OSR1, immunoblotting with phospho-NKCC1 antibodies, Ca2+ imaging in hypothalamic cultures The Journal of neuroscience Medium 22238094
2005 Six1 and Six4 transcription factors directly bind multiple sites in the Slc12a2 promoter (gel-retardation assay) and regulate its expression; in Six1-/-/Six4-/- mice, Slc12a2 expression is reduced in developing dorsal root ganglia, establishing Six1/Six4 as direct transcriptional regulators of NKCC1. Gel-retardation (EMSA) assay, in situ hybridization in Six1/Six4 double-knockout mice The FEBS journal Medium 15955062
2007 NKCC1 is required for NGF-induced neurite outgrowth in PC12D cells: NGF increases NKCC1 protein expression, RNAi knockdown drastically diminishes neurite outgrowth, and EGFP-NKCC1 localizes to the plasma membrane at growth cones during outgrowth. RNAi knockdown, EGFP-tagged live imaging, Western blotting in PC12D cells Biochemical and biophysical research communications Medium 17548052
2007 In developing retinal neurons (ganglion and amacrine cells), NKCC1 does not accumulate intracellular Cl-. GABA-evoked Ca2+ responses persist in NKCC1-null retinas and after bumetanide, and intracellular Cl- is unchanged (~30 mM) in NKCC1-null retinas. Co-staining indicates NKCC1 at P3 is restricted to Müller glia, suggesting NKCC1 buffers extracellular Cl- in Müller cells rather than setting neuronal Cl-. NKCC1-null mice, Ca2+ imaging (fura-2), Cl- imaging (MEQ), immunocytochemistry Journal of neurophysiology High 17493914
2014 In hippocampal slices, NKCC1 inhibition does not affect extracellular K+ clearance after neuronal activity, whereas Na+/K+-ATPase is the primary driver of post-stimulus K+ removal. NKCC1 does mediate astrocyte swelling in response to elevated [K+]o in primary cultures. Ion-selective microelectrodes in rat hippocampal slices, bumetanide pharmacology, volume imaging in primary astrocyte cultures Glia High 24482245
2007 NKCC1 undergoes Ca2+-dependent internalization, lysosomal degradation, and re-expression at basolateral membranes in human colonic epithelium (a 4-hour cycle). This cycle is blocked by the EGFR kinase inhibitor tyrphostin-AG1478 and cycloheximide. In contrast, cAMP (forskolin) sustains NKCC1 membrane expression without internalization, representing a distinct regulatory mode. Immunolabelling, BCECF/Fura-2/calcein imaging, 86Rb+ uptake, pharmacological inhibitors in human colonic crypts The Journal of physiology Medium 17478539
2013 Aldosterone upregulates NKCC1 protein expression independently of mRNA changes by increasing protein stability (reducing ubiquitination), acting through mineralocorticoid receptors (blocked by eplerenone), as shown by cycloheximide and MG132 experiments in HT-29 cells. Pharmacological inhibitors (eplerenone, cycloheximide, MG132), Western blotting, HT-29 cell line American journal of physiology. Cell physiology Medium 24173102
2019 NKCC1 (SLC12A2) is present in a complex with the leucine transporter LAT1-4F2hc. NKCC1 depletion or deletion enhances LAT1 activity, increases Akt and Erk activation, and activates mTORC1 in cells, colonic organoids, and mouse colon, linking NKCC1-mediated cell volume regulation to suppression of mTORC1-dependent cell mass and proliferation. Co-immunoprecipitation (NKCC1–LAT1 complex), NKCC1 KO/KD, mTORC1 activity assays, organoid and mouse colon models Cell reports High 31067471
2017 The ubiquitin ligase Nedd4L suppresses NKCC1 protein abundance in mouse distal colon. Intestinal epithelium-specific Nedd4L knockout mice show increased NKCC1 protein levels and elevated bumetanide-sensitive short-circuit current; however, no direct Co-IP between Nedd4L and NKCC1 was detected, indicating indirect suppression. Conditional intestinal Nedd4L knockout mice, immunoblotting, short-circuit current (Ussing chamber), co-immunoprecipitation (negative) The Journal of biological chemistry Medium 28087701
2015 N-glycosylation is required for NKCC1 plasma membrane targeting and transport function. Inhibition of the first step of N-glycan biosynthesis (tunicamycin) nearly abolishes plasma membrane NKCC1 and cotransport activity. Inhibition of N-glycan maturation (swainsonine/kifunensine) increases core/hybrid-type NKCC1 but eliminates complex N-glycosylated plasma membrane NKCC1 and transport function. Glycosylation inhibitors (tunicamycin, swainsonine, kifunensine), surface biotinylation, Rb+ transport assay in COS7 cells International journal of cell biology Medium 26351455
2011 N-terminal threonine phosphorylation of both NKCC1 and NKCC2A by the WNK-SPAK/OSR1 kinase axis correlates with, but does not fully predict, transporter activity. Phosphorylation of N-termini establishes transport capacity, but final activity also depends on additional factors, so phospho-antibody readout alone is insufficient to infer activity. Stable HEK-293 expression, 86Rb+ flux, phospho-specific immunoblotting under multiple conditions (low Cl-, ouabain, Na+-free, kinase/phosphatase inhibitors) PloS one Medium 21464992
2020 A gain-of-function missense variant in NKCC1 (p.Y199C) located in the N-terminal regulatory domain increases Cl--dependent and bumetanide-sensitive NKCC1 activity even under hypotonicity (when wild-type NKCC1 is normally inactive), establishing that this residue contributes to activity-state regulation. Heterologous expression, Cl- flux assays under various osmotic conditions, comparison to wild-type NKCC1 in Xenopus oocytes/cells Journal of psychiatric research Medium 26955005
2020 De novo mutations in SLC12A2 reduce co-transporter function as demonstrated in Xenopus laevis oocyte expression assays, and cause a neurodevelopmental disorder and/or bilateral sensorineural hearing loss, establishing loss-of-function as the pathogenic mechanism. Xenopus laevis oocyte expression and flux assay of patient variants, trio exome sequencing Brain High 32658972
2016 A truncating SLC12A2 mutation (p.Val1026Phefs*2) produces a non-functional NKCC1 that traffics to the plasma membrane alongside wild-type protein. Patient-derived fibroblasts show reduced total and NKCC1-mediated K+ influx, and the deficit in NKCC1 regulation is revealed only under hypertonic conditions. No dominant-negative effect on wild-type transporter activity was detected. Heterologous expression, K+ transport assay in patient fibroblasts, bumetanide sensitivity, immunoblotting Cold Spring Harbor molecular case studies Medium 27900370
2021 Choroid plexus (ChP) NKCC1 mediates CSF K+ clearance during postnatal development. ChP-specific AAV-NKCC1 overexpression increases CSF K+ clearance and reduces ventriculomegaly in obstructive hydrocephalus mice. A phosphodeficient NKCC1 (AAV-NKCC1-NT51) fails to mitigate ventriculomegaly, establishing that NKCC1 phosphorylation/activation is required for this CSF clearance function. AAV-mediated ChP-specific NKCC1 overexpression and phosphodeficient mutant, CSF K+ measurement, intracranial pressure monitoring, hydrocephalus mouse model Nature communications High 33469018
2023 In the choroid plexus (ChP), intraventricular blood raises CSF K+ and triggers cytosolic Ca2+ activity in ChP epithelial cells, activating NKCC1. ChP-targeted AAV-NKCC1 prevents blood-induced ventriculomegaly; phosphodeficient AAV-NKCC1-NT51 fails to mitigate ventriculomegaly, confirming that NKCC1 phosphorylation is required for trans-choroidal CSF K+ clearance. Intraventricular blood injection model, Ca2+ imaging in ChP, AAV gene therapy with phosphodeficient mutant, ventriculomegaly measurement in mice Neuron High 36893755
2022 Microglial NKCC1 regulates baseline and reactive microglia morphology, process recruitment to injury sites, cell volume adaptation, and membrane conductance in a cell-autonomous manner. Microglial NKCC1 deficiency results in NLRP3 inflammasome priming and increased IL-1β production, and microglial NKCC1 KO mice show increased brain injury and inflammation after experimental stroke. Microglia-specific NKCC1 conditional knockout mouse, morphology imaging, patch-clamp, NLRP3/IL-1β assays, experimental stroke model PLoS biology High 35085235
2017 NKCC1 promotes an EMT-like process in glioblastoma by facilitating GTP-loading of Rac1 and RhoA. Pharmacological inhibition or knockdown of NKCC1 decreases mesenchymal markers (N-cadherin, vimentin, snail) and attenuates activated Rac1/RhoA, while Rac1/RhoA inhibitors impair glioma invasion, placing NKCC1 upstream of these GTPases. shRNA knockdown, pharmacological inhibition, GTP-Rac1/RhoA pulldown assay, invasion assays, intracranial mouse model Journal of cellular physiology Medium 30159893
2020 In NGLY1-deficient mouse cells, NKCC1 shows altered average molecular weight and reduced function, suggesting NGLY1-mediated deglycosylation is required for normal NKCC1 processing and activity, identified through a Drosophila modifier screen (Ncc69/NKCC1) and validated in mammalian cells. Drosophila genetic modifier screen, NKCC1 molecular weight and functional assay in NGLY1-/- mouse cells eLife Medium 33315011
2017 NKCC1 deficiency in goblet cells impairs mucus granule exocytosis, leading to secretion of intact granules into the colonic lumen. NKCC1-DFX (truncation) or complete NKCC1 loss causes aggravated inflammatory response to Citrobacter rodentium infection and decreased expression of claudin-2, implicating NKCC1-dependent ion/water transport in gut barrier function. NKCC1-DFX knock-in mouse model, electron microscopy, immunostaining, FISH, Citrobacter infection model, multiplex cytokine assay Cellular and molecular gastroenterology and hepatology High 31655271
2020 TRPV1 activation by capsaicin causes rapid NKCC1 phosphorylation and increased NKCC1-dependent Rb+ uptake (via Akt) in lens epithelial cells; TRPV1-/- cells show no NKCC1 phosphorylation or Rb+ uptake response, establishing a TRPV1→Akt→NKCC1 activation pathway for osmotic homeostasis in the lens. TRPV1 knockout mice, Rb+ uptake assay, phospho-NKCC1 immunoblotting, hydrostatic pressure measurement, TRPV1 agonist/antagonist pharmacology American journal of physiology. Cell physiology Medium 32293931
2020 Staurosporine and NEM dephosphorylate NKCC1 at Thr203, Thr207, and Thr212, and dephosphorylate the SPAK T-loop (Thr233) and S-loop (Ser373), demonstrating that the reciprocal regulation of NKCC1 (and KCC2) by these agents is mediated through the WNK-SPAK/OSR1 signaling module acting on specific phospho-sites. Mass spectrometry, phospho-specific immunoblotting in HEK293 cells and hippocampal neurons PloS one Medium 32413057
2019 HIF-1α positively regulates NKCC1 transcription via hypoxia-responsive element (HRE) motifs in the promoter, while NFAT5 negatively regulates NKCC1 transcription via tonicity enhancer elements (TonE). Mutation of HRE motifs or pharmacological HIF-1α inhibition reduces hypoxia-induced NKCC1 upregulation; NFAT5 knockdown or TonE mutation increases NKCC1 expression under normal conditions. HIF-1α inhibition, NFAT5 knockdown, promoter-reporter constructs with HRE/TonE mutations, ChIP, Western blotting in hippocampal neurons Frontiers in cell and developmental biology Medium 31921851
2005 NKCC1 is abundantly expressed on the basolateral plasma membrane of secretory coil cells (not apical membrane or epidermis) in rat, mouse, and human eccrine sweat glands, as confirmed by immunoelectron microscopy; this basolateral localization accounts for bumetanide-sensitive NaCl secretion. RT-PCR, immunoblotting, immunohistochemistry, immunoelectron microscopy in rat/mouse/human sweat gland tissue American journal of physiology. Cell physiology High 15843440
2023 NKCC1 protein is expressed at high levels in oligodendrocytes, at lower levels in microglia, astrocytes, developing pericytes, and progenitor cells of the dentate gyrus. In immature neurons, NKCC1 protein localizes to somata; in adult neurons only NKCC1 mRNA is detectable. A differential splice-variant expression was identified: NKCC1a predominates in non-neuronal cells, NKCC1b in neurons. KO-validated immunohistochemistry with custom antibodies, advanced mRNA approaches, single-cell analysis in mouse brain Cerebral cortex High 36573432

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2001 CCC1 is a transporter that mediates vacuolar iron storage in yeast. The Journal of biological chemistry 272 11390404
2011 Expression of GABA signaling molecules KCC2, NKCC1, and GAD1 in cortical development and schizophrenia. The Journal of neuroscience : the official journal of the Society for Neuroscience 257 21795557
2014 Contributions of the Na⁺/K⁺-ATPase, NKCC1, and Kir4.1 to hippocampal K⁺ clearance and volume responses. Glia 222 24482245
2003 PASK (proline-alanine-rich STE20-related kinase), a regulatory kinase of the Na-K-Cl cotransporter (NKCC1). The Journal of biological chemistry 220 12740379
2012 Cation-chloride cotransporters NKCC1 and KCC2 as potential targets for novel antiepileptic and antiepileptogenic treatments. Neuropharmacology 213 22705273
1997 Expression of the Na(+)-K(+)-2Cl- cotransporter BSC2 in the nervous system. The American journal of physiology 202 9038823
1999 Mutation of the Na-K-Cl co-transporter gene Slc12a2 results in deafness in mice. Human molecular genetics 138 10401008
2012 SPAK/OSR1 regulate NKCC1 and WNK activity: analysis of WNK isoform interactions and activation by T-loop trans-autophosphorylation. The Biochemical journal 114 22032326
1998 Comparison of Na-K-Cl cotransporters. NKCC1, NKCC2, and the HEK cell Na-L-Cl cotransporter. The Journal of biological chemistry 110 9556622
2019 Structure and mechanism of the cation-chloride cotransporter NKCC1. Nature 100 31367042
2021 Choroid plexus NKCC1 mediates cerebrospinal fluid clearance during mouse early postnatal development. Nature communications 96 33469018
2010 Altered expression of regulators of the cortical chloride transporters NKCC1 and KCC2 in schizophrenia. Archives of general psychiatry 92 20819979
2020 Role of NKCC1 and KCC2 in Epilepsy: From Expression to Function. Frontiers in neurology 89 32010056
2008 NKCC1 and KCC2 prevent hyperexcitability in the mouse hippocampus. Epilepsy research 80 18394864
2008 NKCC1-mediated traumatic brain injury-induced brain edema and neuron death via Raf/MEK/MAPK cascade. Critical care medicine 80 18431281
1996 The role of the Saccharomyces cerevisiae CCC1 gene in the homeostasis of manganese ions. Molecular microbiology 78 8866476
2021 The structural basis of function and regulation of neuronal cotransporters NKCC1 and KCC2. Communications biology 73 33597714
1997 Partial cloning and characterization of Slc12a2: the gene encoding the secretory Na+-K+-2Cl- cotransporter. The American journal of physiology 70 9357771
2020 NKCC1, an Elusive Molecular Target in Brain Development: Making Sense of the Existing Data. Cells 64 33291778
2000 CCC1 suppresses mitochondrial damage in the yeast model of Friedreich's ataxia by limiting mitochondrial iron accumulation. The Journal of biological chemistry 63 10713071
2007 Dynamic and differential regulation of NKCC1 by calcium and cAMP in the native human colonic epithelium. The Journal of physiology 62 17478539
2016 NKCC1 up-regulation contributes to early post-traumatic seizures and increased post-traumatic seizure susceptibility. Brain structure & function 59 27586142
2020 Structure of the human cation-chloride cotransporter NKCC1 determined by single-particle electron cryo-microscopy. Nature communications 58 32081947
1997 Expression of the bumetanide-sensitive Na-K-Cl cotransporter BSC2 is differentially regulated by fluid mechanical and inflammatory cytokine stimuli in vascular endothelium. The Journal of clinical investigation 56 9185518
2023 Choroid plexus-targeted NKCC1 overexpression to treat post-hemorrhagic hydrocephalus. Neuron 55 36893755
2005 Contribution of the basolateral isoform of the Na-K-2Cl- cotransporter (NKCC1/BSC2) to renin secretion. American journal of physiology. Renal physiology 54 16106034
2016 Astaxanthin alleviates cerebral edema by modulating NKCC1 and AQP4 expression after traumatic brain injury in mice. BMC neuroscience 53 27581370
2021 Pharmacological tools to target NKCC1 in brain disorders. Trends in pharmacological sciences 52 34620512
2005 Hypotension in NKCC1 null mice: role of the kidneys. American journal of physiology. Renal physiology 52 16159893
2020 NKCC1: Newly Found as a Human Disease-Causing Ion Transporter. Function (Oxford, England) 50 33345190
2017 NKCC1-Mediated GABAergic Signaling Promotes Postnatal Cell Death in Neocortical Cajal-Retzius Cells. Cerebral cortex (New York, N.Y. : 1991) 48 26819276
2015 NKCC1 and NKCC2: The pathogenetic role of cation-chloride cotransporters in hypertension. Genes & diseases 48 26114157
2009 Nkcc1 (Slc12a2) is required for the regulation of endolymph volume in the otic vesicle and swim bladder volume in the zebrafish larva. Development (Cambridge, England) 47 19633174
2020 SLC12A2 variants cause a neurodevelopmental disorder or cochleovestibular defect. Brain : a journal of neurology 45 32658972
2017 Astaxanthin protects astrocytes against trauma-induced apoptosis through inhibition of NKCC1 expression via the NF-κB signaling pathway. BMC neuroscience 44 28490320
2014 Cellular iron homeostasis mediated by the Mrs4-Ccc1-Smf3 pathway is essential for mitochondrial function, morphogenesis and virulence in Candida albicans. Biochimica et biophysica acta 44 24368185
2016 A patient with multisystem dysfunction carries a truncation mutation in human SLC12A2, the gene encoding the Na-K-2Cl cotransporter, NKCC1. Cold Spring Harbor molecular case studies 43 27900370
2014 NKCC1 mediates traumatic brain injury-induced hippocampal neurogenesis through CREB phosphorylation and HIF-1α expression. Pflugers Archiv : European journal of physiology 41 25201604
2007 Conservation of hearing by simultaneous mutation of Na,K-ATPase and NKCC1. Journal of the Association for Research in Otolaryngology : JARO 41 17674100
2022 Structural basis for inhibition of the Cation-chloride cotransporter NKCC1 by the diuretic drug bumetanide. Nature communications 40 35585053
2007 NKCC1 does not accumulate chloride in developing retinal neurons. Journal of neurophysiology 40 17493914
2023 Expression patterns of NKCC1 in neurons and non-neuronal cells during cortico-hippocampal development. Cerebral cortex (New York, N.Y. : 1991) 39 36573432
2023 Cation-Chloride Cotransporters KCC2 and NKCC1 as Therapeutic Targets in Neurological and Neuropsychiatric Disorders. Molecules (Basel, Switzerland) 39 36771011
2018 NKCC1 promotes EMT-like process in GBM via RhoA and Rac1 signaling pathways. Journal of cellular physiology 39 30159893
2019 The Ion Transporter NKCC1 Links Cell Volume to Cell Mass Regulation by Suppressing mTORC1. Cell reports 38 31067471
2002 Molecular regulation of Na+-K+-2Cl- cotransporter (NKCC1) and epithelial chloride secretion. World journal of surgery 38 11948363
2018 Pretreatment with AQP4 and NKCC1 Inhibitors Concurrently Attenuated Spinal Cord Edema and Tissue Damage after Spinal Cord Injury in Rats. Frontiers in physiology 37 29403391
2019 Kilquist syndrome: A novel syndromic hearing loss disorder caused by homozygous deletion of SLC12A2. Human mutation 36 30740830
2008 Expression of aquaporin1, 3, and 4, NKCC1, and NKCC2 in the human endolymphatic sac. Auris, nasus, larynx 36 18606512
2016 Gain-of-function missense variant in SLC12A2, encoding the bumetanide-sensitive NKCC1 cotransporter, identified in human schizophrenia. Journal of psychiatric research 35 26955005
2013 Direct control of Na(+)-K(+)-2Cl(-)-cotransport protein (NKCC1) expression with aldosterone. American journal of physiology. Cell physiology 35 24173102
2011 Phosphorylation and transport in the Na-K-2Cl cotransporters, NKCC1 and NKCC2A, compared in HEK-293 cells. PloS one 35 21464992
2007 Essential role of NKCC1 in NGF-induced neurite outgrowth. Biochemical and biophysical research communications 34 17548052
2005 Slc12a2 is a direct target of two closely related homeobox proteins, Six1 and Six4. The FEBS journal 34 15955062
2019 Sevoflurane-Induced Dysregulation of Cation-Chloride Cotransporters NKCC1 and KCC2 in Neonatal Mouse Brain. Molecular neurobiology 32 31493242
2020 A Drosophila screen identifies NKCC1 as a modifier of NGLY1 deficiency. eLife 31 33315011
2005 NKCC1 and NHE1 are abundantly expressed in the basolateral plasma membrane of secretory coil cells in rat, mouse, and human sweat glands. American journal of physiology. Cell physiology 31 15843440
2014 BDNF modifies hippocampal KCC2 and NKCC1 expression in a temporal lobe epilepsy model. Acta neurobiologiae experimentalis 30 25231847
2003 Expression and regulation of the Na+-K+-2Cl- cotransporter NKCC1 in the normal and CFTR-deficient murine colon. The Journal of physiology 30 12692180
1996 Culturing induced expression of basolateral Na+-K+-2Cl- cotransporter BSC2 in proximal tubule, aortic endothelium, and vascular smooth muscle. Pflugers Archiv : European journal of physiology 29 8584442
2022 The NKCC1 ion transporter modulates microglial phenotype and inflammatory response to brain injury in a cell-autonomous manner. PLoS biology 28 35085235
2020 Activated WNK3 induced by intracerebral hemorrhage deteriorates brain injury maybe via WNK3/SPAK/NKCC1 pathway. Experimental neurology 28 32589890
2012 Kinases SPAK and OSR1 are upregulated by estradiol and activate NKCC1 in the developing hypothalamus. The Journal of neuroscience : the official journal of the Society for Neuroscience 28 22238094
2021 Restoring neuronal chloride homeostasis with anti-NKCC1 gene therapy rescues cognitive deficits in a mouse model of Down syndrome. Molecular therapy : the journal of the American Society of Gene Therapy 26 34058387
2017 The Ubiquitin Ligase Nedd4L Regulates the Na/K/2Cl Co-transporter NKCC1/SLC12A2 in the Colon. The Journal of biological chemistry 26 28087701
2017 Discovery of NKCC1 as a potential therapeutic target to inhibit hepatocellular carcinoma cell growth and metastasis. Oncotarget 26 29029515
2016 WNK-Cab39-NKCC1 signaling increases the susceptibility to ischemic brain damage in hypertensive rats. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 26 27798271
2013 DISC1 and SLC12A2 interaction affects human hippocampal function and connectivity. The Journal of clinical investigation 26 23921125
1994 Sequence, mapping and disruption of CCC1, a gene that cross-complements the Ca(2+)-sensitive phenotype of csg1 mutants. Yeast (Chichester, England) 26 7941738
2022 Role of SPAK-NKCC1 signaling cascade in the choroid plexus blood-CSF barrier damage after stroke. Journal of neuroinflammation 25 35413993
2020 TRPV1 activation stimulates NKCC1 and increases hydrostatic pressure in the mouse lens. American journal of physiology. Cell physiology 25 32293931
2019 Novel Human NKCC1 Mutations Cause Defects in Goblet Cell Mucus Secretion and Chronic Inflammation. Cellular and molecular gastroenterology and hepatology 25 31655271
2022 Cryo-EM structure of the human NKCC1 transporter reveals mechanisms of ion coupling and specificity. The EMBO journal 24 36239040
2019 Suppression of WNK1-SPAK/OSR1 Attenuates Bone Cancer Pain by Regulating NKCC1 and KCC2. The journal of pain 24 31085334
2015 Impact of Hybrid and Complex N-Glycans on Cell Surface Targeting of the Endogenous Chloride Cotransporter Slc12a2. International journal of cell biology 24 26351455
2021 Inhibition of the NKCC1/NF-κB Signaling Pathway Decreases Inflammation and Improves Brain Edema and Nerve Cell Apoptosis in an SBI Rat Model. Frontiers in molecular neuroscience 23 33867933
2018 Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways. Frontiers in immunology 23 30271405
2022 NF-κB Signaling-Mediated Activation of WNK-SPAK-NKCC1 Cascade in Worsened Stroke Outcomes of Ang II-Hypertensive Mice. Stroke 22 35272484
2021 NKCC1 promotes proliferation, invasion and migration in human gastric cancer cells via activation of the MAPK-JNK/EMT signaling pathway. Journal of Cancer 22 33391422
2021 High-Resolution Views and Transport Mechanisms of the NKCC1 and KCC Transporters. Journal of molecular biology 22 34022207
2020 Structure and function of the vacuolar Ccc1/VIT1 family of iron transporters and its regulation in fungi. Computational and structural biotechnology journal 22 33304466
2017 The glucose sensor Snf1 and the transcription factors Msn2 and Msn4 regulate transcription of the vacuolar iron importer gene CCC1 and iron resistance in yeast. The Journal of biological chemistry 22 28760824
2020 Acute Hyperglycemia Aggravates Lung Injury via Activation of the SGK1-NKCC1 Pathway. International journal of molecular sciences 21 32645929
2022 Inhibition mechanism of NKCC1 involves the carboxyl terminus and long-range conformational coupling. Science advances 20 36306358
2022 The role of KCC2 and NKCC1 in spinal cord injury: From physiology to pathology. Frontiers in physiology 20 36589461
2017 Iron toxicity in yeast: transcriptional regulation of the vacuolar iron importer Ccc1. Current genetics 20 29043483
2014 Ammonia, like K(+), stimulates the Na(+), K(+), 2 Cl(-) cotransporter NKCC1 and the Na(+),K(+)-ATPase and interacts with endogenous ouabain in astrocytes. Neurochemical research 20 24929663
2002 Expression and regulation of the Na(+)/K(+)/2Cl(-) cotransporter NKCC1 in rat liver and human HuH-7 hepatoma cells. Archives of biochemistry and biophysics 20 12054469
2020 Blockade of Cell Volume Regulatory Protein NKCC1 Increases TMZ-Induced Glioma Apoptosis and Reduces Astrogliosis. Molecular cancer therapeutics 19 32393472
2001 Expression and role of sodium, potassium, chloride cotransport (NKCC1) in mouse inner medullary collecting duct (mIMCD-K2) epithelial cells. Pflugers Archiv : European journal of physiology 19 11692276
2020 Staurosporine and NEM mainly impair WNK-SPAK/OSR1 mediated phosphorylation of KCC2 and NKCC1. PloS one 18 32413057
2020 Androgenic Modulation of the Chloride Transporter NKCC1 Contributes to Age-dependent Isoflurane Neurotoxicity in Male Rats. Anesthesiology 18 32930727
2017 The Role of Na:K:2Cl Cotransporter 1 (NKCC1/SLC12A2) in Dental Epithelium during Enamel Formation in Mice. Frontiers in physiology 18 29209227
2022 Screening of Carbofuran-Degrading Bacteria Chryseobacterium sp. BSC2-3 and Unveiling the Change in Metabolome during Carbofuran Degradation. Metabolites 17 35323662
2021 Effects of α7 Nicotinic Acetylcholine Receptor Positive Allosteric Modulator on BDNF, NKCC1 and KCC2 Expression in the Hippocampus following Lipopolysaccharide-Induced Allodynia and Hyperalgesia in a Mouse Model of Inflammatory Pain. CNS & neurological disorders drug targets 17 33380307
2020 Expression of GSK3β, PICK1, NEFL, C4, NKCC1 and Synaptophysin in peripheral blood mononuclear cells of the first-episode schizophrenia patients. Asian journal of psychiatry 16 33373836
2014 Age-related change in the expression of NKCC1 in the cochlear lateral wall of C57BL/6J mice. Acta oto-laryngologica 16 25220727
2020 Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66. Frontiers in physiology 15 32848856
2019 NFAT5 and HIF-1α Coordinate to Regulate NKCC1 Expression in Hippocampal Neurons After Hypoxia-Ischemia. Frontiers in cell and developmental biology 15 31921851
2015 Effects of SPAK knockout on sensorimotor gating, novelty exploration, and brain area-dependent expressions of NKCC1 and KCC2 in a mouse model of schizophrenia. Progress in neuro-psychopharmacology & biological psychiatry 15 25797415

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