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Calmodulin, a highly conserved calcium‐binding protein, plays a crucial role in response to salt stress. Previous studies investigated sequence and function of calmodulin members in some plants, but their roles in rice have not been fully elucidated. Three OsCaM1 genes namely OsCaM1‐1/2/3 encode the same OsCaM1 protein. Here, we found that OsCaM1‐1 had significantly higher expression than the other two genes under salt stress. After 4 weeks of exposure to 75 mM NaCl, OsCaM1‐1 overexpressed mutants showed higher salt tolerance, while knocked‐out mutants exhibited lower salt tolerance, compared to the wild type. Moreover, the oscam1‐1 mutants had higher Na+ concentration and Na+/K+ ratio in both shoots and roots, less instantaneous K+ and Ca2+ fluxes in roots, compared to wild type under salt stress, indicating the involvement of OsCaM1‐1 in regulation of Na+ and K+ homoeostasis via Ca2+ signal. RNA‐seq analysis identified 452 differentially expressed genes (DEGs) regulated by OsCaM1‐1 and salt stress, and they were mainly enriched in nucleus DNA‐binding activities, including ABI5, WRKY76, WRKY48 and bHLH120 transcription factors. Knockout of OsCaM1‐1 also modulated the expression of Na+ transporters, including HKT1;1, HKT1;5, SOS1, NHX1 and NHX4. In conclusion, OsCaM1‐1 positively regulates salt tolerance in rice through mediating ion homoeostasis. Summary statement: Calmodulin plays a crucial role in response to salt stress. Knockout of OsCaM1‐1 modulated the expressions of HKT1;5, HKT1;1, NHX1 and SOS1, highlighting the positive role of OsCaM1‐1 in the regulation of salt tolerance in rice through mediating ion homoeostasis. [ABSTRACT FROM AUTHOR]

Background & Aims: Diarrhea is one of the most common illnesses and is often caused by bacterial infection. Recently, we have shown that human Na+/H+ exchanger NHE3 (hNHE3), but not non-human NHE3s, interacts with the E3 ubiquitin ligase Nedd4-2. We hypothesize that this property of hNHE3 contributes to the increased severity of diarrhea in humans. Methods: We used humanized mice expressing hNHE3 in the intestine (hNHE3int) to compare the contribution of hNHE3 and mouse NHE3 to diarrhea induced by cholera toxin (CTX) and enteropathogenic Escherichia coli (EPEC). We measured Na+/H+ exchange activity and fluid absorption. The role of Nedd4-2 on hNHE3 activity and ubiquitination was determined by knockdown in Caco-2bbe cells. The effects of protein kinase A (PKA), the primary mediator of CTX-induced diarrhea, on Nedd4-2 and hNHE3 phosphorylation and their interaction were determined. Results: The effects of CTX and EPEC were greater in hNHE3int mice than in control wild-type (WT) mice, resulting in greater inhibition of NHE3 activity and increased fluid accumulation in the intestine, the hallmark of diarrhea. Activation of PKA increased ubiquitination of hNHE3 and enhanced interaction of Nedd4-2 with hNHE3 via phosphorylation of Nedd4-2 at S342. S342A mutation mitigated the Nedd4-2–hNHE3 interaction and blocked PKA-induced inhibition of hNHE3. Unlike non-human NHE3s, inhibition of hNHE3 by PKA is independent of NHE3 phosphorylation, suggesting a distinct mechanism of hNHE3 regulation. Conclusions: The effects of CTX and EPEC on hNHE3 are amplified, and the unique properties of hNHE3 may contribute to diarrheal symptoms occurring in humans.

Accumulation of high sodium (Na+) leads to disruption of metabolic processes and decline in plant growth and productivity. Therefore, this study was undertaken to clarify how Na+/H+ exchangers and Na+/K+ transporter genes contribute to Na+ homeostasis and the substantial involvement of lignin biosynthesis genes in salt tolerance in alfalfa (Medicago sativa L.), which is poorly understood. In this study, high Na+ exhibited a substantial reduction of morphophysiological indices and induced oxidative stress indicators in Xingjiang Daye (XJD; sensitive genotype), while Zhongmu (ZM; tolerant genotype) remained unaffected. The higher accumulation of Na+ and the lower accumulation of K+ and K+/(Na+ + K+) ratio were found in roots and shoots of XJD compared with ZM under salt stress. The ZM genotype showed a high expression of SOS1 (salt overly sensitive 1), NHX1 (sodium/hydrogen exchanger 1), and HKT1 (high-affinity potassium transporter 1), which were involved in K+ accumulation and excess Na+ extrusion from the cells compared with XJD. The lignin accumulation was higher in the salt-adapted ZM genotype than the sensitive XJD genotype. Consequently, several lignin biosynthesis-related genes including 4CL2, CCoAOMT, COMT, CCR, C4H, PAL1, and PRX1 exhibited higher mRNA expression in salt-tolerant ZM compared with XJD. Moreover, antioxidant enzyme (catalase, superoxide dismutase, ascorbate peroxidase, and glutathione reductase) activity was higher in ZM relative to XJD. This result suggests that high antioxidant provided the defense against oxidative damages in ZM, whereas low enzyme activity with high Na+ triggered the oxidative damage in XJD. These findings together illustrate the ion exchanger, antiporter, and lignin biosysthetic genes involving mechanistic insights into differential salt tolerance in alfalfa. [ABSTRACT FROM AUTHOR]

Accumulation of high sodium (Na+) leads to disruption of metabolic processes and decline in plant growth and productivity. Therefore, this study was undertaken to clarify how Na+/H+ exchangers and Na+/K+ transporter genes contribute to Na+ homeostasis and the substantial involvement of lignin biosynthesis genes in salt tolerance in alfalfa (Medicago sativa L.), which is poorly understood. In this study, high Na+ exhibited a substantial reduction of morphophysiological indices and induced oxidative stress indicators in Xingjiang Daye (XJD; sensitive genotype), while Zhongmu (ZM; tolerant genotype) remained unaffected. The higher accumulation of Na+ and the lower accumulation of K+ and K+/(Na+ + K+) ratio were found in roots and shoots of XJD compared with ZM under salt stress. The ZM genotype showed a high expression of SOS1 (salt overly sensitive 1), NHX1 (sodium/hydrogen exchanger 1), and HKT1 (high-affinity potassium transporter 1), which were involved in K+ accumulation and excess Na+ extrusion from the cells compared with XJD. The lignin accumulation was higher in the salt-adapted ZM genotype than the sensitive XJD genotype. Consequently, several lignin biosynthesis–related genes including 4CL2, CCoAOMT, COMT, CCR, C4H, PAL1, and PRX1 exhibited higher mRNA expression in salt-tolerant ZM compared with XJD. Moreover, antioxidant enzyme (catalase, superoxide dismutase, ascorbate peroxidase, and glutathione reductase) activity was higher in ZM relative to XJD. This result suggests that high antioxidant provided the defense against oxidative damages in ZM, whereas low enzyme activity with high Na+ triggered the oxidative damage in XJD. These findings together illustrate the ion exchanger, antiporter, and lignin biosysthetic genes involving mechanistic insights into differential salt tolerance in alfalfa.

Asian cultivated rice shows allelic variation in sodium transporter, OsHKT1;5, correlating with shoot sodium exclusion (salinity tolerance). These changes map to intra/extracellularly-oriented loops that occur between four transmembrane-P loop-transmembrane (MPM) motifs in OsHKT1;5. HKT1;5 sequences from more recently evolved Oryza species (O. sativa / O. officinalis complex species) contain two expansions that involve two intracellularly oriented loops/helical regions between MPM domains, potentially governing transport characteristics, while more ancestral HKT1;5 sequences have shorter intracellular loops. We compared homology models for homoeologous OcHKT1;5-K and OcHKT1;5 - L from halophytic O. coarctata to identify complementary amino acid residues in OcHKT1;5-L that potentially enhance affinity for Na+. Using haplotyping, we showed that Asian cultivated rice accessions only have a fraction of HKT1;5 diversity available in progenitor wild rice species (O. nivara and O. rufipogon). Progenitor HKT1;5 haplotypes can thus be used as novel potential donors for enhancing cultivated rice salinity tolerance. Within Asian rice accessions, 10 non-synonymous HKT1;5 haplotypic groups occur. More HKT1;5 haplotypic diversities occur in cultivated indica gene pool compared to japonica. Predominant Haplotypes 2 and 10 occur in mutually exclusive japonica and indica groups, corresponding to haplotypes in O. sativa salt-sensitive and salt-tolerant landraces, respectively. This distinct haplotype partitioning may have originated in separate ancestral gene pools of indica and japonica , or from different haplotypes selected during domestication. Predominance of specific HKT1 ; 5 haplotypes within the 3 000 rice dataset may relate to eco-physiological fitness in specific geo-climatic and/or edaphic contexts. [ABSTRACT FROM AUTHOR]

Kidneys continuously filter an enormous amount of sodium and adapt kidney Na+ reabsorption to match Naþ intake to maintain circulatory volume and electrolyte homeostasis. Males (M) respond to high-salt (HS) diet by translocating proximal tubule Na+/H+ exchanger isoform 3 (NHE3) to the base of the microvilli, reducing activated forms of the distal NaCl cotransporter (NCC) and epithelial Na+ channel (ENaC). Males (M) and females (F) on normal-salt (NS) diet present sex-specific profiles of "transporters" (cotransporters, channels, pumps, and claudins) along the nephron, e.g., F exhibit 40% lower NHE3 and 200% higher NCC abundance than M. We tested the hypothesis that adaptations to HS diet along the nephron will, likewise, exhibit sexual dimorphisms. C57BL/6J mice were fed for 15 days with 4% NaCl diet (HS) versus 0.26% NaCl diet (NS). On HS, M and F exhibited normal plasma [Na+] and [K+], similar urine volume, Na+, K+, and osmolal excretion rates normalized to body weight. In F, like M, HS lowered abundance of distal NCC, phosphorylated NCC, and cleaved (activated) forms of ENaC. The adaptations associated with achieving electrolyte homeostasis exhibit sex-dependent and independent mechanisms. Sex differences in baseline "transporters" abundance persist during HS diet, yet the fold changes during HS diet (normalized to NS) are similar along the distal nephron and collecting duct. Sex-dependent differences observed along the proximal tubule during HS show that female kidneys adapt differently from patterns reported in males, yet achieve and maintain fluid and electrolyte homeostasis. [ABSTRACT FROM AUTHOR]

The roles of clock components in salt stress tolerance remain incompletely characterized in rice. Here, we show that, among OsPRR (Oryza sativa Pseudo‐Response Regulator) family members, OsPRR73 specifically confers salt tolerance in rice. Notably, the grain size and yield of osprr73 null mutants were significantly decreased in the presence of salt stress, with accumulated higher level of reactive oxygen species and sodium ions. RNA sequencing and biochemical assays identified OsHKT2;1, encoding a plasma membrane‐localized Na+ transporter, as a transcriptional target of OsPRR73 in mediating salt tolerance. Correspondingly, null mutants of OsHKT2;1 displayed an increased tolerance to salt stress. Immunoprecipitation‐mass spectrometry (IP‐MS) assays further identified HDAC10 as nuclear interactor of OsPRR73 and co‐repressor of OsHKT2;1. Consistently, H3K9ac histone marks at OsHKT2;1 promoter regions were significantly reduced in osprr73 mutant. Together, our findings reveal that salt‐induced OsPRR73 expression confers salt tolerance by recruiting HDAC10 to transcriptionally repress OsHKT2;1, thus reducing cellular Na+ accumulation. This exemplifies a new molecular link between clock components and salt stress tolerance in rice. SYNOPSIS: The circadian clock regulates abiotic stress tolerance in plants. Here, rice clock component OsPRR73 is shown to confer salt tolerance via chromatin modifications reducing sodium influx transporter OsHKT2;1 expression. OsPRR73 expression is transcriptionally induced by salt stress.osprr73 null mutants exhibit increased sensitivity to salt stress.OsPRR73 expression inhibits OsHKT2;1 transcription.OsPRR73 reduces histone H3K9 acetylation at the OsHKT2;1 promoter by recruiting histone deacetylase HDAC10.OsHKT2;1 loss enhances rice salt tolerance by reducing sodium ion and ROS accumulation. [ABSTRACT FROM AUTHOR]

In 1986, Vladimir Skulachev and his colleagues coined the term "Sodium World" for the group of diverse organisms with sodium (Na)-based bioenergetics. Albeit only few such organisms had been discovered by that time, the authors insightfully noted that "the great taxonomic variety of organisms employing the Na-cycle points to the ubiquitous distribution of this novel type of membrane-linked energy transductions". Here we used tools of bioinformatics to follow expansion of the Sodium World through the evolutionary time and taxonomic space. We searched for those membrane protein families in prokaryotic genomes that correlate with the use of the Na-potential for ATP synthesis by different organisms. In addition to the known Na-translocators, we found a plethora of uncharacterized protein families; most of them show no homology with studied proteins. In addition, we traced the presence of Na-based energetics in many novel archaeal and bacterial clades, which were recently identified by metagenomic techniques. The data obtained support the view that the Na-based energetics preceded the proton-dependent energetics in evolution and prevailed during the first two billion years of the Earth history before the oxygenation of atmosphere. Hence, the full capacity of Na-based energetics in prokaryotes remains largely unexplored. The Sodium World expanded owing to the acquisition of new functions by Na-translocating systems. Specifically, most classes of G-protein-coupled receptors (GPCRs), which are targeted by almost half of the known drugs, appear to evolve from the Na-translocating microbial rhodopsins. Thereby the GPCRs of class A, with 700 representatives in human genome, retained the Na-binding site in the center of the transmembrane heptahelical bundle together with the capacity of Na-translocation. Mathematical modeling showed that the class A GPCRs could use the energy of transmembrane Na-potential for increasing both their sensitivity and selectivity. Thus, GPCRs, the largest protein family coded by human genome, stem from the Sodium World, which encourages exploration of other Na-dependent enzymes of eukaryotes. [ABSTRACT FROM AUTHOR]

Lithium is widely used in the treatment of bipolar affective disorders, but often causes nephrogenic diabetes insipidus (NDI), a condition characterized by a severe urinary concentrating defect. Lithium-induced NDI is associated with dysregulation of the amiloride-sensitive epithelial sodium channel (ENaC), which is essential for renal sodium reabsorption. Sex hormones have been shown to affect the expression of aquaporin-2 (AQP2) and sodium transporters. Therefore, we evaluated whether tamoxifen (TAM), a selective estrogen receptor modulator (SERM), would affect lithium-induced dysregulation of ENaC subunits and natriuresis. Rats were fed with lithium-containing food for 2 weeks to induce NDI and natriuresis. TAM was administered daily via gastric gavage after 1 week of lithium administration. Lithium treatment alone resulted in increased urinary sodium excretion and significant reduction of βENaC and γENaC at both RNA and protein levels. In addition, the plasma sodium level reduced after lithium treatment. Administration of TAM prevented increased urinary sodium excretion as well as attenuated the downregulation of βENaC and γENaC. Consistent with these findings, immunohistochemistry (IHC) showed stronger labeling of βENaC and γENaC subunits in the apical domain of the collecting duct cells in the cortical tissue of lithium-fed rats treated with TAM. Other major sodium transporters including NaPi-2, NKCC2, Na/K-ATPase, and NHE3, are believed not to have an effect on the increased urinary sodium excretion since their expression increased or was unchanged after treatment with lithium. In conclusion, the results demonstrated that TAM rescued the adverse effects of the lithium-induced increase in fractional excretion of sodium after the establishment of lithium-induced NDI.

Lithium is widely used in the treatment of bipolar affective disorders, but often causes nephrogenic diabetes insipidus (NDI), a condition characterized by a severe urinary concentrating defect. Lithium-induced NDI is associated with dysregulation of the amiloride-sensitive epithelial sodium channel (ENaC), which is essential for renal sodium reabsorption. Sex hormones have been shown to affect the expression of aquaporin-2 (AQP2) and sodium transporters. Therefore, we evaluated whether tamoxifen (TAM), a selective estrogen receptor modulator (SERM), would affect lithium-induced dysregulation of ENaC subunits and natriuresis. Rats were fed with lithium-containing food for 2 weeks to induce NDI and natriuresis. TAM was administered daily via gastric gavage after 1 week of lithium administration. Lithium treatment alone resulted in increased urinary sodium excretion and significant reduction of βENaC and γENaC at both RNA and protein levels. In addition, the plasma sodium level reduced after lithium treatment. Administration of TAM prevented increased urinary sodium excretion as well as attenuated the downregulation of βENaC and γENaC. Consistent with these findings, immunohistochemistry (IHC) showed stronger labeling of βENaC and γENaC subunits in the apical domain of the collecting duct cells in the cortical tissue of lithium-fed rats treated with TAM. Other major sodium transporters including NaPi-2, NKCC2, Na/K-ATPase, and NHE3, are believed not to have an effect on the increased urinary sodium excretion since their expression increased or was unchanged after treatment with lithium. In conclusion, the results demonstrated that TAM rescued the adverse effects of the lithium-induced increase in fractional excretion of sodium after the establishment of lithium-induced NDI. [ABSTRACT FROM AUTHOR]

Hypertension can originate from pre- and post-natal insults. High-fat (HF) diet and prenatal dexamethasone (DEX) exposure are both involved in hypertension of developmental origins. We examined whether postnatal HF diet sex-specifically increases the vulnerability to prenatal DEX exposure-induced programmed hypertension in adult offspring. Additionally, we sought to identify candidate proteins involved in programmed hypertension through a mass spectrometry-based quantitative proteomic approach. Male and female offspring were studied separately: control, DEX, HF, and DEX + HF (n=8/group). Pregnant Sprague-Dawley rats received dexamethasone (0.1 mg/kg body weight) or vesicle from gestational day 16-22. Offspring received high-fat diet (D12331, Research Diets) or regular diet from weaning to 4 months of age. Rats were sacrificed at 4 months of age. We found that postnatal HF diet increased vulnerability of prenatal DEX-induced hypertension in male but not in female adult offspring. Additionally, HF and DEX elicited renal programming in a sex-specific fashion. In males, DEX + HF increased renal parvalbumin (PVALB) and carbonic anhydrase III (CA III) protein levels. While prenatal DEX down-regulated PVALB and CA III protein abundance in female offspring kidneys. Moreover, DEX + HF increased renal protein level of type 3 sodium hydrogen exchanger (NHE3) in males but not in females. In conclusion, postnatal HF diet and prenatal DEX exposure synergistically induced programmed hypertension in male-only offspring. DEX + HF induced sex-specific alterations of protein profiles in offspring kidneys. By identifying candidate proteins underlying sex-specific mechanisms, our results could lead to novel offspring sex-specific interventions to prevent hypertension induced by antenatal corticosteroids and postnatal HF intake in both sexes. [ABSTRACT FROM AUTHOR]

Commercial strawberry, Fragaria x ananassa Duch., is a species sensitive to salinity. Under saline conditions, Na + uptake by the plant is increased, while K + uptake is significantly reduced. Maintaining an adequate K + /Na + cytosolic ratio determines the ability of the plant to survive in saline environments. The goal of the present work was to clone and functionally characterize the genes AKT1 and HKT1 involved in K + and Na + transport in strawberry and to determine the relationship of these genes with the responses of three Fragaria spp. genotypes having different ecological adaptations to salt stress. FaHKT1 and FcHKT1 proteins from F. x ananassa and F. chiloensis have 98.1% of identity, while FaAKT1 and FcAKT1 identity is 99.7%. FaHKT1 and FaAKT1 from F. x ananassa , were functionally characterized in Xenopus oocytes. FaHKT1, belongs to the group I of HKT transporters and is selective for Na + . Expression of FaAKT1 in oocytes showed that the protein is a typical inward-rectifying and highly K + -selective channel. The relative expression of Fragaria HKT1 and AKT1 genes was studied in roots of F. x ananassa cv. Camarosa and of F. chiloensis (accessions Bau and Cucao) grown under salt stress. The expression of AKT1 was transiently increased in ʻCamarosaʼ, decreased in ʻCucaoʼ and was not affected in ʻBauʼ upon salt stress. HKT1 expression was significantly increased in roots of ʻCucaoʼ and was not affected in the other two genotypes. The increased relative expression of HKT1 and decreased expression of AKT1 in ʻCucaoʼ roots correlates with the higher tolerance to salinity of this genotype in comparison with ʻCamarosaʼ and ʻBauʼ. [ABSTRACT FROM AUTHOR]

Chronic kidney disease (CKD) is becoming a global burden, despite recent advances in management. CKD can begin in early life by so-called "developmental programming" or "developmental origins of health and disease" (DOHaD). Early-life insults cause structural and functional changes in the developing kidney, which is called renal programming. Epidemiological and experimental evidence supports the proposition that early-life adverse events lead to renal programming and make subjects vulnerable to developing CKD and its comorbidities in later life. In addition to low nephron endowment, several mechanisms have been proposed for renal programming. The DOHaD concept opens a new window to offset the programming process in early life to prevent the development of adult kidney disease, namely reprogramming. Here, we review the key themes on the developmental origins of CKD.We have particularly focused on the following areas: evidence from human studies support fetal programming of kidney disease; insight from animal models of renal programming; hypothetical mechanisms of renal programming; alterations of renal transcriptome in response to early-life insults; and the application of reprogramming interventions to prevent the programming of kidney disease. [ABSTRACT FROM AUTHOR]

There is a consensus that soil salinization causes decreased agricultural production. Several plants may adapt to survive under high salt stress wherein glycophytes fail to grow. Among transporters in plants, the sodium transporter, also known as high-affinity K+- transporter (HKT1) that comes under the HKT gene family, is involved in uptake of sodium through the roots and its recirculation from shoot to root. In the present study, we have examined the role of transporter HKT1 in soybean plants upon addition of 200 mM NaCl and treatment with Pseudomonas sp. strain AK-1. It is reported that HKT1 is permeable to K+/Na+ and systemically alleviates salinity stress through upregulation of gene expression in shoots and down-regulation in roots. Higher transcription levels in shoot recirculate Na+ from shoot xylem to root phloem whereas lower transcription levels in root do not allow sodium to enter the plants through root cells. We have also examined role of exopolysaccharide (EPS) produced by strain AK-1 which helps in the binding of free Na+ from soil and thus makes Na+ unavailable to the soybean plants. Strain with EPS showed decrease in electrical conductivity of soil from 1.1 to 0.9 dS/m in the presence of 200 mM NaCl. In conclusion, treatment with Pseudomonas sp. strain AK-1 exhibits significant rise in shoot/root length, number of lateral roots, shoot/root fresh weight and decreased Na+/K+ ratio under salinity stress. [ABSTRACT FROM AUTHOR]

The regulation of Na-K-ATPase in various tissues is under the control of a number of hormones and peptides that exert both short- and long-term control over its activity. The present study was performed to investigate the effect of chronic insulin treatment on Na-K-ATPase in renal proximal tubular cells. Incubation of opossum kidney (OK) cells, transfected with the rat Na-K-ATPase α1-subunit, with 1 nmol/l insulin for 48 h decreased Na-K-ATPase activity. Insulin decreased α1-protein content and increased α1-serine phosphorylation and α1-adaptor protein 2 (AP2) interaction. Removal of the 26 NH2-terminal (-NT) amino acid from the α1-subunit containing serine/threonine sites abolished the insulinmediated serine phosphorylation and inhibition of Na-K-ATPase. Substitution of serine 16 and 23 with alanine showed a comparable effect on -NT. Insulin increased the activity of protein kinase C (PKC), which was blocked by the phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin. Both PI3K and PKC inhibitors abolished the insulin-mediated inhibition of Na-K-ATPase. Insulin increased the expression of PKC-β1, -δ, -ξ and-λ; however, only PKC-ξ/λ-specific inhibitors blocked insulin-induced phosphorylation and inhibition of Na-K-ATPase. Our data demonstrate that insulin activates the atypical PKC isoforms-ξ/λ via the PI3K pathway. PKC-ξ/λ-induced phosphorylation of the α1-subunit at serine 16 and 23 leads to AP2 recruitment, degradation, and a decrease in Na-K-ATPase activity. [ABSTRACT FROM AUTHOR]

A plant's ability to cope with salt stress is highly correlated with their ability to reduce the accumulation of sodium ions in the shoot. Arabidopsis mutants affected in the ABSCISIC ACID INSENSITIVE ( ABI) 4 gene display increased salt tolerance, whereas ABI4-overexpressors are hypersensitive to salinity from seed germination to late vegetative developmental stages. In this study we demonstrate that abi4 mutant plants accumulate lower levels of sodium ions and higher levels of proline than wild-type plants following salt stress. We show higher HKT1;1 expression in abi4 mutant plants and lower levels of expression in ABI4-overexpressing plants, resulting in reduced accumulation of sodium ions in the shoot of abi4 mutants. HKT1;1 encodes a sodium transporter which is known to unload sodium ions from the root xylem stream into the xylem parenchyma stele cells. We have shown recently that ABI4 is expressed in the root stele at various developmental stages and that it plays a key role in determining root architecture. Thus ABI4 and HKT1;1 are expressed in the same cells, which suggests the possibility of direct binding of ABI4 to the HKT1;1 promoter. In planta chromatin immunoprecipitation and in vitro electrophoresis mobility shift assays demonstrated that ABI4 binds two highly related sites within the HKT1;1 promoter. These sites, GC(C/G) GCTT(T), termed ABI4-binding element ( ABE), have also been identified in other ABI4-repressed genes. We therefore suggest that ABI4 is a major modulator of root development and function. [ABSTRACT FROM AUTHOR]

Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid metabolite formed by phosphorylation of sphingosine. S1P has been indicated to play a significant role in the cardiovascular system. It has been shown that the enzymes for S1P metabolism are expressed in the kidneys. The present study characterized the expression of S1P receptors in the kidneys and determined the role of S1P in the control of renal hemodynamics and sodium excretion. Real-time RT-PCR analyses showed that S1P receptors S1P1, S1P2, and S1P3 were most abundantly expressed in the renal medulla. Immunohistochemistry revealed that all three types of S1P receptors were mainly located in collecting ducts. Intramedullary infusion of FTY720, an S1P agonist, produced a dramatic increase in sodium excretion by twofold and a small but significant increase in medullary blood flow (16%). Administration of W146, an S1P1 antagonist, into the renal medulla blocked the effect of FTY720 and decreased the sodium excretion by 37% when infused alone. The antagonists of S1P2 and S1P3 had no effect. FTY720 produced additive natriuretic effects in combination with different sodium transporter inhibitors except amiloride, an epithelial sodium channel blocker. In the presence of nitric oxide synthase inhibitor l-NAME, FTY720 still increased sodium excretion. These data suggest that S1P produces natriuretic effects via activation of S1P1 in the renal medulla and this natriuretic effect may be through inhibition of epithelial sodium channel, which is nitric oxide independent. It is concluded that S1P is a novel diuretic factor in the renal medulla and may be an important regulator of sodium homeostasis. [ABSTRACT FROM AUTHOR]

We tested the effects of inflammation on renal dopamine D1 receptor signaling cascade, a key pathway that maintains sodium homeostasis and blood pressure during increased salt intake. Inflammation was produced by administering lipopolysaccharide (LPS; 4 mg/kg ip) to rats provided without (normal salt) and with 1% NaCl in drinking water for 2 wk (high salt). Control rats had saline injection and received tap water. We found that LPS increased the levels of inflammatory cytokines, interleukin-6, and tumor necrosis factor-α in the rats given either normalor high-salt intake. Also, these rats had higher levels of oxidative stress markers, malondialdehyde and nitrotyrosine, and lower levels of antioxidant enzyme superoxide dismutase in the renal proximal tubules (RPTs). The nuclear levels of transcription factors NP-κB increased and Nrf2 decreased in the RPTs in response to LPS in rats given normal and high salt. Furthermore, D1 receptor numbers, D1 receptor proteins, and D1 receptor agonist (SKF38393)-mediated 35S-GTPγS binding decreased in the RPTs in these rats. The basal activities of Na-K-ATPase in the RPTs were similar in control and LPS-treated rats given normal and high salt. SKF38393 caused inhibition of Na-K-ATPase activity in the primary cultures of RPTs treated with vehicle but not in the cultures treated with LPS. Furthermore, LPS caused an increase in blood pressure in the rats given high salt but not in the rats given normal salt. These results suggest that LPS differentially regulates NF-κB and Nrf2, produces inflammation, decreases antioxidant enzyme, increases oxidative stress, and causes D1 receptor dysfunction in the RPTs. The LPS-induced dysfunction of renal D1 receptors alters salt handling and causes hypertension in rats during salt overload. [ABSTRACT FROM AUTHOR]

Intracellular Na+ ([Na+]i) regulation plays a crucial role in the structural, mechanical, and electrical properties of myocardium. It is assumed that the [Na+]i handling system may differ not only between normal and diseased hearts but also regionally within a heart. To gain new insight concerning disease- and region-dependent differences in the [Na+]i-regulatory system, we investigated mRNA expression of Na+ transporters, the principal determinants of [Na+]i. Nonischemic pressure-overloaded hypertrophy was created by suprarenal abdominal aortic constriction of 50% for 7 weeks. mRNA abundances of Na+-Ca2+ exchanger (NCX1), Na+-H+ exchanger (NHE1), Na+-K+-2Cl− exchanger (NKCC1) and Na+, K+-ATPase multigene family(α1, α2, α3, and β1 isoforms) were measured by the real-time quantitative polymerase chain reaction method. mRNA abundance of all transporters mediating Na+ influx (NCX1, NHE1, and NKCC1) was significantly upregulated as compared to normal. In contrast, Na+-efflux-mediating transporter (Na+, K+-ATPase) mRNA expression was unaltered between normal and hypertrophic hearts. Losartan, an angiotensin II AT1 receptor antagonist, significantly attenuated upregulation of Na+-influx-mediating transporters induced by aortic constriction. The onset of Na+-influx-mediating transporter upregulation occurred within 5 days following constriction. In normal and hypertrophied hearts, mRNA of all Na+-influx-mediating transporters was expressed in order of abundance as: apex > septum ∼ free wall of left ventricles. A transmural gradient in expression was also evident in normal hearts (midcardium > endo- and epicardium), which was attenuated under hypertrophic development. Myocardial hypertrophy is associated with significant changes in the spatial distribution and expression levels of Na+ transporters. The upregulation of Na influx transporters during hypertrophy may contribute to the remodeling process, modulate contractility and promote arrhythmias. [ABSTRACT FROM AUTHOR]

Abstract: We show that MDCK I cells express, besides the classical (Na+ +K+)ATPase, a Na+-stimulated ATPase activity with the following characteristics: (1) K 0.5 for Na+ 7.5±1.5mM and V max 23.12±1.1nmol Pi/mg per min; (2) insensitive to 1mM ouabain and 30mM KCl; and (3) inhibited by furosemide and vanadate (IC50 42.1±8.0 and 4.3±0.3μM, respectively). This enzyme forms a Na+-stimulated, furosemide- and hydroxylamine-sensitive ATP-driven acylphosphate phosphorylated intermediate with molecular weight of 100kDa. Immunoprecipitation of the (Na+ +K+)ATPase with monoclonal anti-α1 antibody reduced its activity in the supernatant by 90%; the Na+-ATPase activity was completely maintained. In addition, the formation of the Na+-stimulated, furosemide- and hydroxylamine-sensitive ATP-driven acylphosphate intermediate occurred at the same magnitude as that observed before immunoprecipitation. These data suggest that Na+-ATPase and (Na+ +K+)ATPase activities are independent, with Na+-ATPase belonging to a different enzyme entity. [Copyright &y& Elsevier]

Thyrotoxicosis is a common disorder causing cardiovascular and renal irregularities. In this study, thyrotoxicosis was produced in rats by 14 days of daily thyroxine injection. This was associated with an increase in cardiac index, mean arterial pressure, and renal blood flow compared with euthyroid controls. Food and water intake along with urine output were significantly increased in the thyrotoxic rats compared with control animals associated with a significant increase in solute excretion. Polyuria and increased solute excretion still occurred even when food and water intake was equivalent. These renal responses were associated with significant decreases in AQP1 and AQP2 water channel expression in both the ad lib and paired intake studies in the cortex and inner medulla. The downregulation of AQP2 protein occurred in spite of equivalent plasma arginine vasopressin (AVP) in the ad lib and increased AVP in the paired feeding studies. Solute-free water reabsorption was greater in both the ad lib and paired thyrotoxic than euthyroid rats and was associated with increased Na–K–2Cl cotransporter expression. We propose that the AVP-independent downregulation of AQP2, the observed increase in renal arterial pressure, and decrease in filtration fraction contribute to polyuria the increased solute excretion in spite of enhanced ion transporters in thyrotoxicosis.Kidney International (2007) 72, 1088–1094; doi:10.1038/sj.ki.5002475; published online 15 August 2007 [ABSTRACT FROM AUTHOR]

Angiotensin II (ANG II) plays an important role in the development of obstructive nephropathy. Here, we examined the effects of the ANG II receptor type 1 (AT1R) blockade using candesartan on long-term renal molecular and functional changes in response to partial unilateral ureteral obstruction (PUUO). Newborn rats were subjected to severe PUUO or sham operation (Sham) within the first 48 h of life. Candesartan was provided in the drinking water (10 mg·kg-1·day-1) from day 21 of life until 10 wk of age. Renal blood flow (RBF) was evaluated by MRI, glomerular filtration rate (GFR) was measured using the renal clearance of 51Cr-EDTA, and the renal expression of Na-K-ATPase and the collecting duct water channel aquaporin-2 (AQP2) was examined by immunoblotting and immunocytochemistry. At 10 wk of age, PUUO significantly reduced RBF (0.8 ± 0.1 vs. 1.6 ± 0.1 ml·min-1·100 g body wt-1 P < 0.05) and GFR (37 ± 16 vs. 448 ± 111 µl·min-1·100 g body wt-1; P < 0.05) compared with Sham. Candesartan prevented the RBF reduction (PUUO+CAN: 1.6 ± 0.2 vs. PUUO: 0.8 ± 0.1 ml·min-1·100 g body wt-1; P < 0.05) and attenuated the GFR reduction (PUUO+CAN: 265 ± 68 vs. PUUO: 37 ± 16 µl·min-1·100 g body wt-1; P < 0.05). PUUO was also associated with a significant downregulation in the expression of Na-K-ATPase (75 ± 12 vs. 100 ± 5%, P < 0.05) and AQP2 (52 ± 15 vs. 100 ± 4%, P < 0.05), which were also prevented by candesartan (Na-K-ATPase: 103 ± 8 vs. 100 ± 5% and AQP2: 74 ± 13 vs. 100 ± 4%). These findings were confirmed by immunocytochemistry. Consistent with this, candesartan treatment partly prevented the reduction in solute free water reabsorption and attenuated fractional sodium excretion in rats with PUUO. In conclusion, candesartan prevents or attenuates the reduction in RBF, GFR and dysregulation of AQP2 and Na-K-ATPase in response to congenital PUUO in rats, suggesting that AT1R blockade may protect the neonatally obstructed kidney against development of obstructive nephropathy. [ABSTRACT FROM AUTHOR]

Neonatal ureteral obstruction alters expression of renal sodium transporters and aquaporin water channels. Background. Congenital urinary tract obstruction is a common cause of renal insufficiency in the neonate and during infancy. Recently, we demonstrated that ureteral obstruction in adult rats is associated with reduction in the abundance of renal aquaporins (AQPs) and renal sodium transporters, which paralleled an impaired urinary concentrating capacity. Methods. In the present study, renal handling of sodium and water, together with the expression of renal aquaporins and major renal sodium transporters, was examined in rats with neonatally induced partial unilateral ureteral obstruction (PUUO) within the first 48 hours of life to clarify the molecular mechanisms involved in the tubular functional defects in response to congenital obstruction. Rats were then followed for 12 or 24 weeks. Results. Neonatal PUUO caused a progressive reduction in single kidney glomerular filtration rate (SKGFR) on the obstructed side to 43% of controls at 12 weeks (115 ± 28 vs. 267 ± 36 μL/min/100g bw, P < 0.05), and 31% of controls at 24 weeks (106 ± 24 vs. 343 ± 41 μL/min/100g bw, P < 0.05). Na-K-ATPase abundance was decreased in the obstructed kidney compared with the nonobstructed kidney at 24 weeks (79 ± 6%, P < 0.05), and the abundance of bumetanide-sensitive Na-K-2Cl cotransporter (BSC-1) located to the medullary thick ascending limb (mTAL) of the obstructed kidney was significantly reduced both at 12 weeks (42 ± 10%, P < 0.05) and 24 weeks (50 ± 10%, P < 0.05). Immunohistochemistry confirmed down-regulation of BSC-1 both at 12 and 24 weeks after onset of obstruction. Consistent with this, sodium excretion from the obstructed kidney was increased at 12 weeks (0.13 ± 0.03 vs. 0.04 ± 0.01 μmol/min/100g bw, P < 0.05), and persisted 24 weeks after onset of PUUO (0.15 ± 0.02 vs. 0.06 ± 0.01 μmol/min/100g bw, P < 0.05). AQP2 abundance in the collecting duct was also reduced both at 12 weeks (68 ± 5%, P < 0.05) and at 24 weeks (69 ± 13%, P < 0.05). Consistent with this, solute-free water reabsorption was decreased in the obstructed kidney at 12 weeks (0.61 ± 0.42 vs. 1.97 ± 0.63 μL/min/100g bw, P < 0.05) and remained decreased after 24 weeks of PUUO (0.42 ± 0.04 vs. 1.56 ± 0.39 μL/min/100g bw, P < 0.05). Conclusion. Major sodium transporters and aquaporins in the obstructed kidney are down-regulated in response to neonatally induced PUUO, which indicates that these transporters may play a crucial role for the persistent reduction in renal handling of sodium and water in response to PUUO. [ABSTRACT FROM AUTHOR]

Differential upregulation of rat Na-K-Cl cotransporter, rBSC1, mRNA in the thick ascending limb of Henle in different pathological conditions. Background. Na-Cl cotransport across the apical membrane of epithelial cells in the thick ascending limb of the loop of Henle (TAL) plays a major role in salt accumulation for hypertonic medullary interstitium. The electroneutral, rat bumetanide-sensitive sodium transporter, rBSC1, is involved in this process. We studied the level of rBSC1 mRNA in dehydration and cardiac failure, since sodium transport in TAL may be enhanced in both conditions in spite of the difference in extracellular fluid accumulation. Methods. Male Sprague-Dawley rats were deprived of water for 24 hours and myocardial infarction of about 40% of left ventricular circumference was induced in another group of rats that later developed congestive heart failure (CHF). Digoxigenin-labeled cRNA probe for rBSC1 was constructed using polymerase chain reaction (PCR), and Northern blot analysis was performed using RNAs from renal outer medulla. By inducing a point mutation at the middle of PCR product, we compared the amount of rBSC1 transcripts in the renal cortex using competitive PCR, since TAL represents a small fraction of the total cortical tissue. Results. Northern analysis showed a significant increase in rBSC1 mRNA in the renal outer medulla of both dehydrated and CHF rats. In the renal cortex, however, the increase was noted only in CHF by competitive PCR. In situ hybridization using the riboprobe for northern analysis demonstrated that the transcript signal in dehydrated rats was intensified segmentally in TAL located in the inner stripe of outer medulla. Western analysis and immunohistochemistry using a specific antibody against rBSC1 confirmed the distinct segmental enhancement of apical protein expression in dehydration and diffuse enhanced expression in CHF. Conclusions. rBSC1 is differentially upregulated in different pathological conditions.... [ABSTRACT FROM AUTHOR]

• Six HKT transporters have been linked to grapevine leaf Na+ exclusion. • Three HKT1 members remained uncharacterised. • Our voltage clamp experiment identified their different Na+ transport properties. • Existing RNAseq data showed that they were weakly expressed. • Our results support that HKT1;1 plays the major role in grapevine Na+ exclusion. Grapevine (Vitis vinifera L.) is a valuable crop for human consumption and wine production, and is prone to suffering from salinity stress in arid regions or when exposed to low quality irrigation water. A previous study identified a quantitative trait locus (QTL) NaE , containing six High-affinity Potassium Transporter 1 genes, that was associated with shoot Na+ exclusion in grapevine. While HKT1;1 was predicted to be the most likely gene within this QTL to encode for this important salinity tolerance sub-trait, four other HKT s within the QTL remained uncharacterised; VviHKT1;2 encodes a truncated transcript unlikely to form a functional transporter. In this study, two allelic variants for each of VviHKT1;6, VviHKT1;7 and VviHKT1;8 from the heterozygous grapevine variety Cabernet Sauvignon were functionally characterised. Using the Xenopus laevis oocyte heterologous expression system, as well as transient expression in tobacco leaves, we found that the VviHKT1;6 and VviHKT1;7 alleles encoded plasma membrane localised proteins that facilitated significant non-rectifying Na+ transport. Conversely, proteins encoded by the VviHKT1;8 alleles were inwardly-rectifying, weak Na+ transporters that localised to intracellular organelles. Mining of previous RNA-seq gene expression data suggested that VviHKT1;6–8 are weakly expressed in grapevine roots, flower buds, and seeds under normal conditions and different nutrient regimes. We propose that VviHKT1;6 and VviHKT1;7 are likely to have a less significant role in grapevine leaf Na+ exclusion than VviHKT1;1, and that VviHKT1;8 is involved in endomembrane Na+ transport. [ABSTRACT FROM AUTHOR]

Hypertension affects approximately 1.13 billion adults worldwide and is the leading global risk factor for cardiovascular, cerebrovascular, and kidney diseases. There is emerging evidence that extracellular vesicles participate in the development and progression of hypertension. Extracellular vesicles are membrane-enclosed structures released from nearly all types of eukaryotic cells. During their formation, extracellular vesicles incorporate various parent cell components, including proteins, lipids, and nucleic acids that can be transferred to recipient cells. Extracellular vesicles mediate cell-to-cell communication in a variety of physiological and pathophysiological processes. Therefore, studying the role of circulating and urinary extracellular vesicles in hypertension has the potential to identify novel noninvasive biomarkers and therapeutic targets of different hypertension phenotypes. This review discusses the classification and biogenesis of three EV subcategories (exosomes, microvesicles, and apoptotic bodies) and provides a summary of recent discoveries in the potential impact of extracellular vesicles on hypertension with a specific focus on their role in the blood pressure regulation by organs-artery and kidney, as well as renin-angiotensin-system.

The Plasmodium falciparum ATPase Pf ATP4 is the target of a diverse range of antimalarial compounds, including the clinical drug candidate cipargamin. Pf ATP4 was originally annotated as a Ca 2+ transporter, but recent evidence suggests that it is a Na + efflux pump, extruding Na + in exchange for H + Here we demonstrate that ATP4 proteins belong to a clade of P-type ATPases that are restricted to apicomplexans and their closest relatives. We employed a variety of genetic and physiological approaches to investigate the ATP4 protein of the apicomplexan Toxoplasma gondii , Tg ATP4. We show that Tg ATP4 is a plasma membrane protein. Knockdown of Tg ATP4 had no effect on resting pH or Ca 2+ but rendered parasites unable to regulate their cytosolic Na + concentration ([Na + ] cyt ). Pf ATP4 inhibitors caused an increase in [Na + ] cyt and a cytosolic alkalinization in WT but not Tg ATP4 knockdown parasites. Parasites in which Tg ATP4 was knocked down or disrupted exhibited a growth defect, attributable to reduced viability of extracellular parasites. Parasites in which Tg ATP4 had been disrupted showed reduced virulence in mice. These results provide evidence for ATP4 proteins playing a key conserved role in Na + regulation in apicomplexan parasites., (© 2019 Lehane et al.)

The antimalarial activity of chemically diverse compounds, including the clinical candidate cipargamin, has been linked to the ATPase PfATP4 in the malaria-causing parasite Plasmodium falciparum The characterization of PfATP4 has been hampered by the inability thus far to achieve its functional expression in a heterologous system. Here, we optimized a membrane ATPase assay to probe the function of PfATP4 and its chemical sensitivity. We found that cipargamin inhibited the Na + -dependent ATPase activity present in P. falciparum membranes from WT parasites and that its potency was reduced in cipargamin-resistant PfATP4-mutant parasites. The cipargamin-sensitive fraction of membrane ATPase activity was inhibited by all 28 of the compounds in the "Malaria Box" shown previously to disrupt ion regulation in P. falciparum in a cipargamin-like manner. This is consistent with PfATP4 being the direct target of these compounds. Characterization of the cipargamin-sensitive ATPase activity yielded data consistent with PfATP4 being a Na + transporter that is sensitive to physiologically relevant perturbations of pH, but not of [K + ] or [Ca 2+ ]. With an apparent K m for ATP of 0.2 mm and an apparent K m for Na + of 16-17 mm, the protein is predicted to operate at below its half-maximal rate under normal physiological conditions, allowing the rate of Na + efflux to increase in response to an increase in cytosolic [Na + ]. In membranes from a cipargamin-resistant PfATP4-mutant line, the apparent K m for Na + is slightly elevated. Our study provides new insights into the biochemical properties and chemical sensitivity of an important new antimalarial drug target., (© 2018 Rosling et al.)

Hypertension can originate from early-life insults, whereas maternal melatonin therapy can be protective in a variety of models of programmed hypertension. We hypothesize that melatonin or melatonin receptor agonist agomelatine can prevent programmed hypertension in adult offspring induced by maternal exposure to continuous light. Female Sprague-Dawley pregnant rats randomly divided into four groups: controls, rats exposed to continuous light, exposed to continuous light plus treated with agomelatine (50 mg/day i.p.), and exposed to continuous light plus treated with 0.01% melatonin in drinking water throughout pregnancy and lactation period. Male offspring (n = 10/group) from three litters were examined at 12 weeks of age. Maternal continuous light exposure-induced hypertension in male offspring, which was prevented by melatonin or agomelatine therapy. Continuous light exposure did not affect melatonin pathway in adult offspring kidney. Genes that belong to the renin-angiotensin system (RAS), sodium transporters, AMP-activated protein kinase pathway, and circadian rhythm were potentially involved in the maternal exposure to continuous light-induced programmed hypertension. Maternal agomelatine therapy decreased Ace expression but increased Agtr2 and Mas1. Maternal melatonin therapy prevented the increases of Slc9a3, Slc12a3, and Atp1a1 expression induced by maternal continuous light exposure. In conclusion, maternal melatonin or agomelatine therapy prevents programmed hypertension induced by maternal exposure to continuous light. Agomelatine and melatonin reprogram the RAS and sodium transporters differentially, to prevent negative programming of continuous light. Our data highlight candidate genes and pathways in renal programming as targets for therapeutic approaches to prevent programmed hypertension caused by early-life disturbance of the circadian rhythm., (© The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)