Molecular cloning and functional characterization of  KCC3, a new K-Cl cotransporter

Race, JoAnne E.; Makhlouf, Fadi N.; Logue, Paul J.; Wilson, Frederick H.; Dunham, Philip B.; Holtzman, Eliezer J.

doi:10.1152/ajpcell.1999.277.6.c1210

Cited by 196 publications

(110 citation statements)

References 35 publications

(56 reference statements)

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“…These transporters are involved in cell proliferation (Shen et al, 2001) and in the electro-neutral movement of ions across the plasma membrane, thus controlling ionic and osmotic homeostasis of various cell types. The gene SLC12A6 is located on chromosome 15q13-14 (Hiki et al, 1999;Mount et al, 1999;Race et al, 1999). Several groups have demonstrated that mutations in SLC12A6 are causative for the recessively inherited Andermann syndrome (ACCPN, OMIM %218000), a severe neurological disorder characterized by agenesis of the corpus callosum, peripheral neuropathy (Howard et al, 2002bUyanik et al, 2006) and psychoses (Filteau et al, 1991).…”

Section: Introductionmentioning

confidence: 99%

Functional Analysis of a Potassium-Chloride Co-Transporter 3 (SLC12A6) Promoter Polymorphism Leading to an Additional DNA Methylation Site

Moser

Ekawardhani

Kumsta

et al. 2008

Neuropsychopharmacol

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The human potassium-chloride co-transporter 3 (KCC3, SLC12A6) is involved in cell proliferation and in electro-neutral movement of ions across the cell membrane. The gene (SLC12A6) is located on chromosome 15q14, a region that has previously shown linkage with bipolar disorder, schizophrenia, rolandic epilepsy, idiopathic generalized epilepsy, autism and attention deficit/hyperactivity disorder. Furthermore, recessively inherited mutations of SLC12A6 cause Andermann syndrome, characterized by agenesis of the corpus callosum, which is associated with peripheral neuropathy and psychoses. Recently, we have demonstrated the association of two G/A promoter polymorphisms of SLC12A6 with bipolar disorder in a case-control study, and familial segregation of the rare variants as well as a trend toward association with schizophrenia. To investigate functional consequences of these polymorphisms, lymphocyte DNA was extracted, bisulfite modified, and subsequently sequenced. To investigate SLC12A6 promoter activity, various promoter constructs were generated and analyzed by luciferase reporter gene assays. We provide evidence that the G-allele showed a significant reduction of reporter gene expression. In human lymphocytes, the allele harboring the rare upstream G nucleotide was found to be methylated at the adjacent C position, possibly accountable for tissue-specific reduction in gene expression in vivo. Here we demonstrate functionality of an SNP associated with psychiatric disease and our results may represent a functional link between genetic variation and an epigenetic modification.

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Section: Introductionmentioning

confidence: 99%

Functional Analysis of a Potassium-Chloride Co-Transporter 3 (SLC12A6) Promoter Polymorphism Leading to an Additional DNA Methylation Site

Moser

Ekawardhani

Kumsta

et al. 2008

Neuropsychopharmacol

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“…Neuron-specific KCC2 is present in the retina and central nervous system, where it plays a crucial role in maintaining low intracellular Cl − and GABAergic synaptic inhibition Rivera et al, 1999;Vardi et al, 2000;Vu et al, 2000). KCC3 shares a 77% overall amino acid identity with human KCC1, and is expressed at highest levels in skeletal muscle, heart, kidney, brain, and lens (Chee et al, 2006;Hiki et al, 1999;Lee et al, 2003;Mount et al, 1999;Pearson et al, 2001;Race et al, 1999). Human KCC4 shares a 69% amino acid identity with KCC2, and is expressed in muscle, heart, kidney, brain, lung, and lens (Chee et al, 2006;Lee et al, 2003;Mercado et al, 2000;Mount et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Functional and molecular characterization of multiple K–Cl cotransporter isoforms in corneal epithelial cells

Capó-Aponte

Wang

Bildin

et al. 2007

Experimental Eye Research

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The dependence of regulatory volume decrease (RVD) activity on potassium-chloride cotransporter (KCC) isoform expression was characterized in corneal epithelial cells (CEC). During exposure to a 50% hypotonic challenge, the RVD response was larger in SV40-immortalized human CEC (HCEC) than in SV40-immortalized rabbit CEC (RCEC). A KCC inhibitor-[(dihydroindenyl)oxy] alkanoic acid (DIOA)-blocked RVD more in HCEC than RCEC. Under isotonic conditions, Nethylmaleimide (NEM) produced KCC activation and transient cell shrinkage. Both of these changes were greater in HCEC than in RCEC. Immunoblot analysis of HCEC, RCEC, primary human CEC (pHCEC), and primary bovine CEC (BCEC) plasma membrane enriched fractions revealed KCC1, KCC3, and KCC4 isoform expression, whereas KCC2 was undetectable. During a hypotonic challenge, KCC1 membrane content increased more rapidly in HCEC than in RCEC. Such a challenge induced a larger increase and more transient p44/42MAPK activation in HCEC than RCEC. On the other hand, HCEC and RCEC p38MAPK phosphorylation reached peak activations at 2.5 and 15 min, respectively. Only in HCEC, pharmacological manipulation of KCC activity modified the hypotonicity-induced activation of p44/42MAPK, whereas p38MAPK phosphorylation was insensitive to such procedures in both cell lines. Larger increases in HCEC KCC1 membrane protein content correlate with their ability to undergo faster and more complete RVD. Furthermore, pharmacological activation of KCC increased p44/42MAPK phosphorylation in HCEC but not in RCEC, presumably a reflection of low KCC membrane expression in RCEC. These findings suggest that KCC1 plays a role in (i) maintaining isotonic steady-state cell volume homeostasis, (ii) recovery of isotonic cell volume after a hypotonic challenge through RVD, and (iii) regulating hypotonicityinduced activation of the p44/42MAPK signaling pathway required for cell proliferation.

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“…They are highly conserved across species. They mediate the uptake of structurally diverse organic acids and some neutral compounds in exchange for intracellular ␣-ketoglutarate (21)(22)(23). Their substrates include many endogenous and exogenous compounds, such as dicarboxylates (20), prostaglandins (24), cyclic nucleotides and urate (16,20,23), nonsteroidal anti-inflammatory drugs (25), diuretics (26), antibiotics (27), and antiviral agents (28).…”

mentioning

confidence: 99%

Uptake of Chemically Reactive, DNA-Damaging Sulfuric Acid Esters into Renal Cells by Human Organic Anion Transporters

Bakhiya

Stephani²,

Bahn³

et al. 2006

Journal of the American Society of Nephrology

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The procarcinogen 1-methylpyrene is activated by hepatic enzymes via 1-hydroxymethylpyrene to 1-sulfooxymethylpyrene (1-SMP), a highly reactive and mutagenic metabolite. Previously, high levels of 1-SMP DNA adducts were observed in rat kidneys after intraperitoneal administration of 1-hydroxymethylpyrene or 1-SMP. This study examined whether organic anion transporters (OAT) that are expressed at the basolateral membrane of proximal tubule cells are involved in uptake of SMP. Human epithelial kidney (HEK293) cells that stably express human OAT1 (hOAT1) and hOAT3 were used. Stable isomers of 1-SMP, (2-SMP and 4-SMP) competitively inhibited the uptake of characteristic substrates p-aminohippurate for hOAT1 and estrone sulfate for hOAT3. Both inhibitors exhibited high affinity for hOAT1 (K i ‫؍‬ 4.4 M for 2-SMP; K i ‫؍‬ 5.1 M for 4-SMP) as well as hOAT3 (K i ‫؍‬ 1.9 M for 2-SMP; K i ‫؍‬ 2.1 M for 4-SMP). The uptake rate of 4-SMP (at a concentration of 10 M) by hOAT1-and hOAT3-expressing cells was 3.0 and 1.6 times higher, respectively, than in control cells. Uptake of the reactive isomer 1-SMP was investigated using as the end point the level of DNA adducts that were formed in the cells. After exposure to 1-SMP (10 M), the DNA adduct level was 4.6 and 3.0 times higher in hOAT1-and hOAT3-expressing cells, respectively, than in control cells. The enhanced DNA adduct formation in hOAT-expressing cells was abolished in the presence of the OAT inhibitor probenecid. This study indicates that OAT can mediate the basolateral uptake of reactive sulfuric acid esters into proximal tubule cells and thereby participate in kidney cell damage by these compounds.

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Molecular cloning and functional characterization of KCC3, a new K-Cl cotransporter

Cited by 196 publications

References 35 publications

Functional Analysis of a Potassium-Chloride Co-Transporter 3 (SLC12A6) Promoter Polymorphism Leading to an Additional DNA Methylation Site

Functional Analysis of a Potassium-Chloride Co-Transporter 3 (SLC12A6) Promoter Polymorphism Leading to an Additional DNA Methylation Site

Functional and molecular characterization of multiple K–Cl cotransporter isoforms in corneal epithelial cells

Uptake of Chemically Reactive, DNA-Damaging Sulfuric Acid Esters into Renal Cells by Human Organic Anion Transporters

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