Stimulation of Na,K-ATPase by low potassium requires reactive oxygen species

2003 ◽  
Vol 285 (2) ◽  
pp. C319-C326 ◽  
Author(s):  
Xiaoming Zhou ◽  
Wu Yin ◽  
Sonia Q. Doi ◽  
Shawn W. Robinson ◽  
Kunio Takeyasu ◽  
...  
Keyword(s):  
Binding Sites ◽  
Regulatory Element ◽  
Protein Abundance ◽  
Oxygen Species ◽  
Ouabain Binding ◽  
Direct Stimulation ◽  
Negative Regulatory Element ◽  
Low K ◽  
Cat Expression ◽  
Stimulation Of

The signaling pathway that transduces the stimulatory effect of low K+ on the biosynthesis of Na,K-ATPase remains largely unknown. The present study was undertaken to examine whether reactive oxygen species (ROS) mediated the effect of low K+ in Madin-Darby canine kidney (MDCK) cells. Low K+ increased ROS activity in a time- and dose-dependent manner, and this effect was abrogated by catalase and N-acetylcysteine (NAC). To determine the role of ROS in low-K+-induced gene expression, the cells were first stably transfected with expression constructs in which the reporter gene chloramphenicol acetyl transferase (CAT) was under the control of the avian Na,K-ATPase α-subunit 1.9 kb and 900-bp 5′-flanking regions that have a negative regulatory element. Low K+ increased the CAT expression in both constructs. Catalase or NAC inhibited the effect of low K+. To determine whether the increased CAT activity was mediated through releasing the repressive effect or a direct stimulation of the promoter, the cells were transfected with a CAT expression construct directed by a 96-bp promoter fragment that has no negative regulatory element. Low K+ also augmented the CAT activity expressed by this construct. More importantly, both catalase and NAC abolished the effect of low K+. Moreover, catalase and NAC also inhibited low-K+-induced increases in the Na,K-ATPase α1- and β1-subunit protein abundance and ouabain binding sites. The antioxidants had no significant effect on the basal levels of CAT activity, protein abundance, or ouabain binding sites. In conclusion, low K+ enhances the Na,K-ATPase gene expression by a direct stimulation of the promoter activity, and ROS mediate this stimulation and also low-K+-induced increases in the Na,K-ATPase protein contents and cell surface molecules.

Excitation-induced activation of the Na(+)-K+ pump in rat skeletal muscle

1994 ◽  
Vol 266 (4) ◽  
pp. C925-C934 ◽  
Author(s):  
M. E. Everts ◽  
T. Clausen
Keyword(s):  
Soleus Muscle ◽  
Contractile Activity ◽  
Activation Mechanism ◽  
Rat Skeletal Muscle ◽  
Ouabain Binding ◽  
Direct Stimulation ◽  
Rat Soleus Muscle ◽  
Fold Stimulation ◽  
86Rb Influx ◽  
Stimulation Of

The stimulating effect of excitation on the Na(+)-K+ pump was characterized in measurements of 22Na efflux, intracellular Na+ content, 86Rb influx, and [3H]ouabain binding in isolated rat soleus muscle. Direct stimulation (10 V, 1 ms, 2 Hz) rapidly increased 22Na efflux and 86Rb influx about twofold. These effects were blocked by tetracaine and ouabain, were not associated with any significant increase in intracellular Na+, and could not be attributed to a rise in extracellular K+. The stimulation of 22Na efflux was unaffected by tubocurarine, dantrolene, trifluoperazine, or bumetanide. Stimulation at 2 Hz increased the rate of [3H]ouabain binding by approximately 120% within 1 min, indicating an early specific activation of the Na(+)-K+ pump. Stimulation at 60 Hz for 10 s increased intracellular Na+ content by 58%. Reextrusion of Na+ was complete in 2 min and could be prevented by ouabain (10(-4) M) or by cooling to 0 degrees C. It is concluded that, in rat soleus muscle, excitation leads to a rapid and pronounced (up to 15-fold) stimulation of the Na(+)-K+ pump, even at modest increases in intracellular Na+. This activation mechanism may be essential for the maintenance of transmembrane Na(+)-K+ gradients and prompt recovery of excitability during contractile activity.

scholarly journals Cerium-Promoted Ginsenosides Accumulation by Regulating Endogenous Methyl Jasmonate Biosynthesis in Hairy Roots of Panax ginseng

Molecules ◽  
2021 ◽  
Vol 26 (18) ◽  
pp. 5623
Author(s):  
Ru Zhang ◽  
Shiquan Tan ◽  
Bianling Zhang ◽  
Pengcheng Hu ◽  
Ling Li
Keyword(s):  
Methyl Jasmonate ◽  
Hairy Roots ◽  
Signal Pathways ◽  
Ros Production ◽  
Oxygen Species ◽  
Direct Stimulation ◽  
Key Enzyme ◽  
Mda Content ◽  
Ginsenoside Content ◽  
Stimulation Of

Among rare earth elements, cerium has the unique ability of regulating the growth of plant cells and the biosynthesis of metabolites at different stages of plant development. The signal pathways of Ce3+-mediated ginsenosides biosynthesis in ginseng hairy roots were investigated. At a low concentration, Ce3+ improved the elongation and biomass of hairy roots. The Ce3+-induced accumulation of ginsenosides showed a high correlation with the reactive oxygen species (ROS), as well as the biosynthesis of endogenous methyl jasmonate (MeJA) and ginsenoside key enzyme genes (PgSS, PgSE and PgDDS). At a Ce3+ concentration of 20 mg L−1, the total ginsenoside content was 1.7-fold, and the total ginsenosides yield was 2.7-fold that of the control. Malondialdehyde (MDA) content and the ROS production rate were significantly higher than those of the control. The activity of superoxide dismutase (SOD) was significantly activated within the Ce3+ concentration range of 10 to 30 mg L−1. The activity of catalase (CAT) and peroxidase (POD) strengthened with the increasing concentration of Ce3+ in the range of 20–40 mg L−1. The Ce3+ exposure induced transient production of superoxide anion (O2•−) and hydrogen peroxide (H2O2). Together with the increase in the intracellular MeJA level and enzyme activity for lipoxygenase (LOX), there was an increase in the gene expression level of MeJA biosynthesis including PgLOX, PgAOS and PgJMT. Our results also revealed that Ce3+ did not directly influence PgSS, PgSE and PgDDS activity. We speculated that Ce3+-induced ROS production could enhance the accumulation of ginsenosides in ginseng hairy roots via the direct stimulation of enzyme genes for MeJA biosynthesis. This study demonstrates a potential approach for understanding and improving ginsenoside biosynthesis that is regulated by Ce3+-mediated signal transduction.

Cholera toxin-sensitive neurons in guinea pig submucosal plexus

1993 ◽  
Vol 264 (1) ◽  
pp. G86-G94 ◽  
Author(s):  
M. M. Jiang ◽  
A. Kirchgessner ◽  
M. D. Gershon ◽  
A. Surprenant
Keyword(s):  
Guinea Pig ◽  
Cholera Toxin ◽  
Binding Sites ◽  
Enteric Neurons ◽  
Nerve Terminals ◽  
Submucosal Plexus ◽  
Direct Stimulation ◽  
Vascular Perfusion ◽  
B Subunit ◽  
Stimulation Of

Cholera toxin (CT) increases intestinal secretions by direct stimulation of mucosal enterocytes; enteric neurons also may play a role. We tested the latter possibility by retrograde labeling of mucosal terminals in guinea pig small intestine with the B subunit of CT (B-CT) and by intracellular recordings from submucosal neurons during superfusion with CT. All vasoactive intestinal peptide (VIP)-positive neurons, and only VIP-positive neurons, were labeled with B-CT. Fluorogold (FG) was used to retrogradely label nerve terminals in submucosal arterioles in preparations in which B-CT labeled mucosal terminals; colocalization of B-CT with FG was observed in neurons up to 3 mm from the site of FG application. CT selectively depolarized neurons known to contain VIP. We conclude that all VIP-containing neurons, and only VIP neurons, in guinea pig submucosal plexus possess B-CT binding sites and can be activated by CT. Some of these neurons provide a dual innervation to both arterioles and mucosa. We suggest that one functional consequence of CT may be to activate vasodilator nerves, thus increasing vascular perfusion of the mucosa to further stimulate intestinal secretions.

scholarly journals Ouabain binding and coupled sodium, potassium, and chloride transport in isolated transverse tubules of skeletal muscle.

1979 ◽  
Vol 74 (3) ◽  
pp. 335-349 ◽  
Author(s):  
Y H Lau ◽  
A H Caswell ◽  
M Garcia ◽  
L Letellier
Keyword(s):  
Binding Sites ◽  
Chloride Transport ◽  
Slow Phase ◽  
Ouabain Binding ◽  
Fast Phase ◽  
Highly Active ◽  
Tubular Lumen ◽  
Number Of Binding Sites ◽  
T Tubules ◽  
Low K

The affinity and number of binding sites of [3H]ouabain to isolated transverse (T) tubules were determined in the absence and presence of deoxycholate. In both conditions the KD was approximately 53 nM while deoxycholate increased the number of binding sites from 3.5 to 37 pmol/mg protein. We concluded that the ouabain binding sites were located primarily on the inside of the isolated vesicle and that the vesicles were impermeable to ouabain. ATP induced a highly active Na+ accumulation by the T tubules which increased Na+ in the T tubular lumen by almost 200 nmol/mg protein. The accumulation had an initial fast phase lasting 2-3 min and a subsequent slow phase which continued for at least 40 min. The rate of the initial fast phase indicated a turnover number of 20 Na+/s. The Na+ accumulation was prevented by monensin but was unaffected by valinomycin. Ouabain did not influence Na+ uptake, but digitoxin inhibited it. At low K+ the accumulation of Na+ was reduced 3.7-fold below the value at 50 mM K+. 86Rb, employed as a tracer to detect K+, showed a first phase of K+ release while Na+ was accumulated. After 2-3 min, K+ was reaccumulated while Na+ continued to increase in the lumen. T tubules accumulated Cl- on addition of ATP. This suggested that ATP initiated an exchange of Na+ for K+ followed by uptake of Na+ and K+ accompanied by Cl-.

scholarly journals Transcriptional Regulation of Genes Encoding the Selenium-Free [NiFe]-Hydrogenases in the Archaeon Methanococcus voltae Involves Positive and Negative Control Elements

Genetics ◽  
1999 ◽  
Vol 152 (4) ◽  
pp. 1335-1341
Author(s):  
Izabela Noll ◽  
Steffen Müller ◽  
Albrecht Klein
Keyword(s):  
Intergenic Region ◽  
Genetic Information ◽  
Regulatory Element ◽  
Transcription Unit ◽  
Negative Control ◽  
Negative Regulation ◽  
Negative Regulatory Element ◽  
Methanococcus Voltae ◽  
Genes Encoding ◽  
Made In

Abstract Methanococcus voltae harbors genetic information for two pairs of homologous [NiFe]-hydrogenases. Two of the enzymes contain selenocysteine, while the other two gene groups encode apparent isoenzymes that carry cysteinyl residues in the homologous positions. The genes coding for the selenium-free enzymes, frc and vhc, are expressed only under selenium limitation. They are transcribed out of a common intergenic region. A series of deletions made in the intergenic region localized a common negative regulatory element for the vhc and frc promoters as well as two activator elements that are specific for each of the two transcription units. Repeated sequences, partially overlapping the frc promoter, were also detected. Mutations in these repeated heptanucleotide sequences led to a weak induction of a reporter gene under the control of the frc promoters in the presence of selenium. This result suggests that the heptamer repeats contribute to the negative regulation of the frc transcription unit.

scholarly journals The Role of BMP Signaling in Osteoclast Regulation

2021 ◽  
Vol 9 (3) ◽  
pp. 24
Author(s):  
Brian Heubel ◽  
Anja Nohe
Keyword(s):  
Bone Formation ◽  
Bone Morphogenetic Proteins ◽  
Bone Diseases ◽  
Bmp Signaling ◽  
Bone Homeostasis ◽  
Direct Stimulation ◽  
Federal Drug Administration ◽  
Bmp Pathway ◽  
Stimulation Of

The osteogenic effects of Bone Morphogenetic Proteins (BMPs) were delineated in 1965 when Urist et al. showed that BMPs could induce ectopic bone formation. In subsequent decades, the effects of BMPs on bone formation and maintenance were established. BMPs induce proliferation in osteoprogenitor cells and increase mineralization activity in osteoblasts. The role of BMPs in bone homeostasis and repair led to the approval of BMP2 by the Federal Drug Administration (FDA) for anterior lumbar interbody fusion (ALIF) to increase the bone formation in the treated area. However, the use of BMP2 for treatment of degenerative bone diseases such as osteoporosis is still uncertain as patients treated with BMP2 results in the stimulation of not only osteoblast mineralization, but also osteoclast absorption, leading to early bone graft subsidence. The increase in absorption activity is the result of direct stimulation of osteoclasts by BMP2 working synergistically with the RANK signaling pathway. The dual effect of BMPs on bone resorption and mineralization highlights the essential role of BMP-signaling in bone homeostasis, making it a putative therapeutic target for diseases like osteoporosis. Before the BMP pathway can be utilized in the treatment of osteoporosis a better understanding of how BMP-signaling regulates osteoclasts must be established.

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