Arginine vasopressin stimulates H+-ATPase in MDCK cells via V1 (cell Ca2+) and V2 (cAMP) receptors

2004 ◽  
Vol 286 (2) ◽  
pp. F402-F408 ◽  
Author(s):  
Maria Oliveira-Souza ◽  
Raif Musa-Aziz ◽  
Gerhard Malnic ◽  
Margarida de Mello Aires
Keyword(s):  
Arginine Vasopressin ◽  
Mdck Cells ◽  
Specific Inhibitor ◽  
Cytosolic Calcium ◽  
Dependent Manner ◽  
Madin Darby Canine Kidney ◽  
Dose Dependent ◽  
Darby Canine Kidney ◽  
Canine Kidney ◽  
Stimulation Of

The effect of arginine vasopressin (AVP) and/or atrial natriuretic peptide (ANP) on the regulation of intracellular pH (pHi) via H+-ATPase and of cytosolic calcium ([Ca2+]i) was investigated in Madin-Darby canine kidney (MDCK) cells by the fluorescent probes BCECF-AM and fluo-4-AM, respectively. The pHi recovery rate was examined after intracellular acidification following an NH4Cl pulse, in the presence of zero Na+ plus Schering 28080 (a specific inhibitor of H+-K+-ATPase). AVP (10-12-10-6 M) increased the rate of pHi recovery and [Ca2+]i in a dose-dependent manner. V1- or V2-receptor antagonists impaired the effect of AVP on both processes, and DDAVP (10-12-10-6 M; a V2-selective agonist) caused a dose-dependent stimulation of them. [Ca2+]i or cAMP (as increased by 10-5 M thapsigargin or 8-BrcAMP, respectively) alone had no effect on H+-ATPase, but their synergic action was necessary to stimulate H+-ATPase. In agreement with these findings, ANP (10-6 M) or dimethyl-BAPTA-AM (5 × 10-5 M), impairing the increase of [Ca2+]i in response to AVP, blocks the stimulatory effect of AVP on H+-ATPase.

scholarly journals Microtubule perturbation inhibits intracellular transport of an apical membrane glycoprotein in a substrate-dependent manner in polarized Madin-Darby canine kidney epithelial cells.

1990 ◽  
Vol 1 (12) ◽  
pp. 921-936 ◽  
Author(s):  
M J van Zeijl ◽  
K S Matlin
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Intracellular Transport ◽  
Mdck Cells ◽  
Sodium Dodecyl ◽  
Apical Plasma Membrane ◽  
Dependent Manner ◽  
Madin Darby Canine Kidney ◽  
Darby Canine Kidney ◽  
Canine Kidney

The effects of microtubule perturbation on the transport of two different viral glycoproteins were examined in infected Madin-Darby canine kidney (MDCK) cells grown on both permeable and solid substrata. Quantitative biochemical analysis showed that the microtubule-depolymerizing drug nocodazole inhibited arrival of influenza hemagglutinin on the apical plasma membrane in MDCK cells grown on both substrata. In contrast, the microtubule-stabilizing drug taxol inhibited apical appearance of hemagglutinin only when MDCK cells were grown on permeable substrata. On the basis of hemagglutinin mobility on sodium dodecyl sulfate gels and its sensitivity to endo H, it was evident that nocodazole and taxol arrested hemagglutinin at different intracellular sites. Neither drug caused a significant increase in the amount of hemagglutinin detected on the basolateral plasma membrane domain. In addition, neither drug had any noticeable effect on the transport of the vesicular stomatitis virus (VSV)-G protein to the basolateral surface. These results shed light on previous conflicting reports using this model system and support the hypothesis that microtubules play a role in the delivery of membrane glycoproteins to the apical, but not the basolateral, domain of epithelial cells.

H(+)-peptide cotransport in Madin-Darby canine kidney cells: expression and calmodulin-dependent regulation

1995 ◽  
Vol 268 (3) ◽  
pp. F391-F397 ◽  
Author(s):  
M. Brandsch ◽  
V. Ganapathy ◽  
F. H. Leibach
Keyword(s):  
Mdck Cells ◽  
Peptide Transport ◽  
Maximal Velocity ◽  
Dependent Manner ◽  
Beta Lactam ◽  
Madin Darby Canine Kidney ◽  
Carbonyl Cyanide ◽  
Lactam Antibiotic ◽  
Darby Canine Kidney ◽  
Canine Kidney

The transport of the dipeptide glycylsarcosine was studied in the kidney cell lines OK, LLC-PK1, and Madin-Darby canine kidney (MDCK), grown as confluent monolayers on impermeable plastic supports. Uptake of the dipeptide in OK and LLC-PK1 cells was slow, was not inhibited by other peptides, and was not influenced by an inwardly directed H+ gradient, indicating lack of expression of the H(+)-peptide cotransport system in these cells under our conditions. In contrast, uptake of the dipeptide in MDCK cells was rapid and was found to be stimulated by an inwardly directed H+ gradient. This stimulation was markedly reduced by the protonophore carbonyl cyanide p-trifluoromethoxy-phenylhydrazone. The H+ gradient-dependent uptake of glycylsarcosine was inhibited by dipeptides and tripeptides and by the beta-lactam antibiotic cephalexin but not by the amino acids glycine and leucine. The uptake was saturable and apparently occurred via a single transport system. The Michaelis-Menten constant for the system was 1.3 +/- 0.1 mM, and the maximal velocity was 13.3 +/- 0.7 nmol.30.min-1.mg protein-1. Treatment of MDCK cells with the calmodulin antagonists N-(6-aminohexyl)-5-chloro-1-napthalenesulfonamide (W-7), CGS-9343B, or calmidazolium inhibited the glycylsarcosine uptake by 40-50% in a time- and dose-dependent manner. In contrast, the uptake of alanine, leucine, glucose, and taurine was found to be stimulated by treatment with W-7. Kinetic analysis revealed that the inhibition of the peptide transport activity was mainly associated with a decrease of the maximal velocity of the system.(ABSTRACT TRUNCATED AT 250 WORDS)

Cycloheximide increases glucocorticoid-stimulated α-ENaC mRNA in collecting duct cells by p38 MAPK-dependent pathway

2003 ◽  
Vol 284 (4) ◽  
pp. F778-F787 ◽  
Author(s):  
Omar A. Itani ◽  
Kristyn L. Cornish ◽  
Kang Z. Liu ◽  
Christie P. Thomas
Keyword(s):  
P38 Mapk ◽  
Gene Transcription ◽  
Collecting Duct ◽  
Dependent Manner ◽  
Madin Darby Canine Kidney ◽  
Collecting Duct Cells ◽  
Mrna Half Life ◽  
Dose Dependent ◽  
Darby Canine Kidney ◽  
Canine Kidney

Aldosterone and glucocorticoids (GCs) stimulate Na+ reabsorption in the collecting ducts by increasing the activity of the epithelial Na+ channel (ENaC). Our laboratory has used Madin-Darby canine kidney-C7 cells to demonstrate that this effect is associated with an increase in α-ENaC gene transcription (Mick VE, Itani OA, Loftus RW, Husted RF, Schmidt TJ, and Thomas CP, Mol Endocrinol 15: 575–588, 2001). Cycloheximide (CHX) superinduced the GC-stimulated α-ENaC expression in a dose-dependent manner, but had no effect on basal or aldosterone-stimulated α-ENaC expression, whereas anisomycin inhibited basal and corticosteroid-stimulated α-ENaC expression. The superinduction of α-ENaC expression was also seen with hypotonicity, was blocked by RU-38486, and was independent of protein synthesis. CHX had no effect on α-ENaC mRNA half-life, confirming that its effect was via an increase in α-ENaC transcription. The effect of CHX and hypotonicity on α-ENaC expression was abolished by SB-202190, indicating an effect mediated via p38 MAPK. Consistent with this scheme, CHX increased pp38 and MKK6, an upstream activator of p38, stimulated α-ENaC promoter activity. These data confirm a model in which CHX activates p38 in Madin-Darby canine kidney-C7 cells to increase α-ENaC gene transcription in a GC-dependent manner.

Effect of Riluzole on Ca2+ Movement and Cytotoxicity in Madin-Darby Canine Kidney Cells

2006 ◽  
Vol 25 (8) ◽  
pp. 461-469 ◽  
Author(s):  
W-C Chen ◽  
H-H Cheng ◽  
C-J Huang ◽  
C-T Chou ◽  
S-I Liu ◽  
...  
Keyword(s):  
Mdck Cells ◽  
Dependent Manner ◽  
Madin Darby Canine Kidney ◽  
Concentration Dependent Manner ◽  
Intracellular Ca ◽  
Pre Treatment ◽  
Lateral Sclerosis ◽  
Darby Canine Kidney ◽  
Canine Kidney

Riluzole is a drug used in the treatment of amyotrophic lateral sclerosis; however, its in vitro action is unclear. In this study, the effect of riluzole on intracellular Ca2- concentration ([Ca2-]i) in Madin-Darby canine kidney (MDCK) cells was investigated using the Ca2--sensitive fluorescent dye, fura-2. Riluzole (100 -500 mM) caused a rapid and sustained increase of [Ca2-]i in a concentration-dependent manner (EC50 = 150 mM). Some 40 and 50% of this [Ca2-]i increase was prevented by the removal of extracellular Ca2- and the addition of La3-, respectively, but was unchanged by dihydropyridines, verapamil and diltiazem. In Ca2--free medium, thapsigargin -an inhibitor of the endoplasmic reticulum (ER) Ca2--ATPase -caused a monophasic [Ca2-]i increase, after which the increasing effect of riluzole on [Ca2-]iwas attenuated by 70%; in addition, pre-treatment with riluzole abolished thapsigargin-induced [Ca2-]i increases. U73122, an inhibitor of phospholipase C (PLC), abolished ATP (but not riluzole)-induced [Ca2-]i increases. At concentrations of 250 and 500 mM, riluzole killed 40 and 95% cells, respectively. The cytotoxic effect of riluzole (250 mM) was unaltered by pre-chelating cytosolic Ca2- with BAPTA. Collectively, in MDCK cells, riluzole rapidly increased [Ca2-]i by stimulating extra-cellular Ca2- influx via an La3--sensitive pathway and intracellular Ca2- release from the ER via, as yet, unidentified mechanisms. Furthermore, riluzole caused Ca2--unrelated cytotoxicity in a concentration-depen-dent manner.

Urea flux across MDCK-mUT-A2 monolayers is acutely sensitive to AVP, cAMP, and [Ca2+]i

2006 ◽  
Vol 291 (1) ◽  
pp. F122-F128 ◽  
Author(s):  
Elizabeth A. Potter ◽  
Gavin Stewart ◽  
Craig P. Smith
Keyword(s):  
Mdck Cells ◽  
Type I ◽  
Urea Transporter ◽  
Madin Darby Canine Kidney ◽  
In Trans ◽  
First Time ◽  
Dose Dependent Increase ◽  
Dose Dependent ◽  
Darby Canine Kidney ◽  
Canine Kidney

In this study, we engineered a Madin-Darby canine kidney (MDCK) type I cell line to stably express the mouse urea transporter UT-A2. Monolayers of MDCK-mUT-A2 cells had a basal phloretin-inhibitable urea permeability of 8.4 × 10−6 ± 0.3 cm/s. Treatment of MDCK-mUT-A2 monolayers with AVP led to a rapid dose-dependent increase in trans-monolayer phloretin-inhibitable urea flux. The temporal pattern of response was markedly different from that observed for MDCK cells expressing rat UT-A1. Exposure of MDCK-mUT-A2 cells to either 10 μM forskolin or 250 μM 8-bromo cAMP also increased urea flux rate. Inclusion of the PKA inhibitor H89 (10 μM) had no effect on the forskolin-stimulated increase in urea flux across MDCK-mUT-A2 monolayers. Treatment with either 10 μM CPA or 1 mM ATP also caused an increase in UT-A2-mediated urea flux, although these responses where transient compared with those induced by AVP or elevated cAMP. Taken together, these results show for the first time that UT-A2 is acutely sensitive to AVP, cAMP, or increased intracellular calcium.

scholarly journals Mechanism of Antiviral Activity of Nitazoxanide against the Influenza Virus: Effect of Tizoxanide on AdenosineTriphosphate in Influenza-virus Infected Madin Darby Canine Kidney Cells

2021 ◽  
Author(s):  
Jean-Francois Rossignol ◽  
Carl van Baalen ◽  
Aloys Tijsma
Keyword(s):  
Influenza Virus ◽  
Influenza A ◽  
Respiratory Infections ◽  
Mdck Cells ◽  
Dependent Manner ◽  
Madin Darby Canine Kidney ◽  
Atp Production ◽  
Viral Respiratory Infections ◽  
Darby Canine Kidney ◽  
Canine Kidney

Background: Nitazoxanide (NTZ) is a broad-spectrum antiviral undergoing clinical development for treating influenza and other viral respiratory infections such as those caused by rhinovirus/enterovirus and coronavirus including the emerging SARS-CoV-2. Methods: Nitazoxanide is a mild uncoupler of oxidative phosphorylation, which is modulating the ATP production in cells. ATP is an essential component of viral replication, and we have evaluated the effect of tizoxanide (TIZ), the active circulating metabolite of NTZ, on ATP in Madin-Darby canine kidney (MDCK) cells and in MDCK cells infected with influenza A and B viruses. Results: TIZ decreased cellular ATP in a dose-dependent manner in MDCK cells and in MDCK cells infected with influenza A and B viruses. Maximum inhibition of ATP in influenza infected or uninfected MDCK cells reached up to 45% after 6 and 24 hours of exposure to 100 micrometer TIZ. The decrease in cellular ATP did not affect cell viability and was reversible after eliminating TIZ from the culture. Conclusion: The concentrations of TIZ required to decrease cellular ATP levels were similar to those reported to inhibit replication of influenza A and B viruses in our laboratory. A decrease in ATP triggers activation of AMP-activated protein kinase, which is known to suppress the secretion of pro-inflammatory cytokines. Additional studies are warranted to evaluate the effect of TIZ on mitochondrial function.

Differential sensitivity of Madin-Darby canine kidney (MDCK) cells to epinephrine

2015 ◽  
Vol 20 (5) ◽  
pp. 486-493 ◽  
Author(s):  
P. Muthuraman ◽  
P. C. Nagajyothi ◽  
M. Chandrasekaran ◽  
G. Enkhtaivan ◽  
B. Venkitasamy ◽  
...  
Keyword(s):  
Mdck Cells ◽  
Differential Sensitivity ◽  
Madin Darby Canine Kidney ◽  
Darby Canine Kidney ◽  
Canine Kidney

Accumulation of natural and synthetic betaines by a mammalian renal cell line

1996 ◽  
Vol 74 (2) ◽  
pp. 283-287 ◽  
Author(s):  
K. Randall ◽  
M. Lever ◽  
B. A. Peddie ◽  
S. T. Chambers
Keyword(s):  
Osmotic Stress ◽  
Glycine Betaine ◽  
Mdck Cells ◽  
Intracellular Accumulation ◽  
Madin Darby Canine Kidney ◽  
Renal Cell Line ◽  
High Concentrations ◽  
Inhibition Studies ◽  
Darby Canine Kidney ◽  
Canine Kidney

Intracellular accumulation of different betaines was compared in osmotically stressed Madin Darby canine kidney (MDCK) cells to model the betaine accumulation specificity of the mammalian inner medulla and to show how this accumulation differed from that of bacteria. All betaines accumulated less than glycine betaine. Arsenobetaine (the arsenic analogue of glycine betaine) accumulated to 12% of the glycine betaine levels and the sulphur analogue dimethylthetin accumulated to >80%. Most substituted glycine betaine analogues accumulated to 2–5% of intracellular glycine betaine concentrations, however, serine betaine accumulated to <0.5% of glycine betaine levels. Inhibition studies to distinguish the betaine ports were performed by the addition of proline. Butyrobetaine and carnitine accumulation was not proline sensitive, whereas that of omer betaines was. As with glycine betaine, the accumulation of propionobetaine and dimethylthetin was proline sensitive and osmoregulated. Pyridinium betaine was accumulated by both proline-sensitive and -insensitive systems, with a small increase under osmotic stress. High concentrations (10 times that of glycine betaine) of the dietary betaines proline betaine and trigonelline inhibited total betaine accumulation. Because α-substituted betaines are accumulated by bacteria and not by MDCK cells, these betaines may be the basis for design of antimicrobial agents.Key words: MDCK cells, betaine accumulation, osmolytes, betaine analogues.

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