Endothelin-1 stimulates NO production and inhibits cAMP accumulation in rat inner medullary collecting duct through independent pathways

2006 ◽  
Vol 290 (6) ◽  
pp. F1315-F1319 ◽  
Author(s):  
Peter K. Stricklett ◽  
Alisa K. Hughes ◽  
Donald E. Kohan
Keyword(s):  
Collecting Duct ◽  
Cell Suspensions ◽  
No Production ◽  
Camp Accumulation ◽  
No Donors ◽  
Endothelin 1 ◽  
Inner Medullary Collecting Duct ◽  
Nos Inhibitors ◽  
Medullary Collecting Duct ◽  
Spermine Nonoate

Endothelin-1 (ET-1) inhibition of vasopressin (AVP)-stimulated cAMP accumulation in the collecting duct has been hypothesized to be mediated, at least in part, by nitric oxide (NO). To examine this, the effect of ET-1 on NO production by acutely isolated rat inner medullary collecting duct (IMCD) cell suspensions and the role of NO in mediating ET-1 effects on AVP-stimulated cAMP accumulation were studied. ET-1 dose dependently (first evident at 100 pM ET-1) increased IMCD NO production as determined by DAF-FM fluorescence. ETB receptor (BQ-788), but not ETA receptor (BQ-123), antagonism blocked this effect. Nonspecific NO synthase (NOS) inhibitors [ NG-nitro-l-arginine methyl ester (l-NAME) or NG-monomethyl-l-arginine] or NOS-1 inhibitors (SMTC or VNIO) inhibited the ET-1 response, whereas NOS-2 or NOS-3 inhibitors (l-NAA or 1400W) were ineffective. ET-1 also increased cGMP accumulation. ET-1 caused a 35% reduction in AVP-stimulated cAMP levels; however, this response was not affected by l-NAME or SMTC. The addition of l-arginine, NADPH, tetrahydrobiopterin, or tempol (to reduce superoxide-dependent conversion of NO to peroxynitrate) did not affect the response. NO donors (SNAP or spermine NONOate), at concentrations that stimulated DAF-FM fluorescence and increased cGMP levels, did not alter AVP-stimulated cAMP accumulation in the IMCD cell suspensions. In conclusion, ET-1 stimulates IMCD NO production through activation of the ETB receptor and NOS-1. However, neither ET-1-mediated NO production nor NO donors inhibit AVP-stimulated cAMP accumulation, indicating that NO does not mediate ET-1 inhibition of cAMP production by the IMCD.

Autocrine role of endothelin in rat IMCD: inhibition of AVP-induced cAMP accumulation

1993 ◽  
Vol 265 (1) ◽  
pp. F126-F129 ◽  
Author(s):  
D. E. Kohan ◽  
A. K. Hughes
Keyword(s):  
Arginine Vasopressin ◽  
Collecting Duct ◽  
Camp Accumulation ◽  
Semipermeable Membranes ◽  
Endothelin 1 ◽  
Cyclic Monophosphate ◽  
Nephron Segment ◽  
Specific Receptors ◽  
Medullary Collecting Duct

Exogenous endothelin-1 (ET-1) inhibits arginine vasopressin (AVP)-induced adenosine 3',5'-cyclic monophosphate (cAMP) accumulation in the inner medullary collecting duct (IMCD). Since ET-1 is produced by, and binds to specific receptors on, the IMCD, the possibility exists that ET-1 is an autocrine regulator of AVP action in this nephron segment. To test this hypothesis, rat IMCD cells grown on semipermeable membranes were exposed to rabbit anti-ET antisera or nonimmune rabbit sera (NRS). AVP (10(-9)M) caused a significantly greater accumulation of cAMP in confluent IMCD monolayers preincubated in ET-1 antisera compared with NRS. ET-1 (10(-8) M) inhibited the AVP-induced rise in cAMP by 65% in cells preincubated in ET-1 antisera, but had no effect in NRS-treated cells. Finally, 125I-ET-1 (30 pM) binding was increased sixfold in IMCD preincubated in anti-ET-1 antisera. These data indicate that ET causes tonic autocrine inhibition of AVP responsiveness in the IMCD.

scholarly journals High salt induces autocrine actions of ET-1 on inner medullary collecting duct NO production via upregulated ETB receptor expression

2016 ◽  
Vol 311 (2) ◽  
pp. R263-R271 ◽  
Author(s):  
Kelly Anne Hyndman ◽  
Courtney Dugas ◽  
Alexandra M. Arguello ◽  
Traci T. Goodchild ◽  
Kathleen M. Buckley ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Collecting Duct ◽  
High Salt ◽  
Receptor Expression ◽  
Electrolyte Balance ◽  
No Production ◽  
Inner Medullary Collecting Duct ◽  
Nitrite Production ◽  
Vasa Recta ◽  
Medullary Collecting Duct

The collecting duct endothelin-1 (ET-1), endothelin B (ETB) receptor, and nitric oxide synthase-1 (NOS1) pathways are critical for regulation of fluid-electrolyte balance and blood pressure control during high-salt feeding. ET-1, ETB receptor, and NOS1 are highly expressed in the inner medullary collecting duct (IMCD) and vasa recta, suggesting that there may be cross talk or paracrine signaling between the vasa recta and IMCD. The purpose of this study was to test the hypothesis that endothelial cell-derived ET-1 (paracrine) and collecting duct-derived ET-1 (autocrine) promote IMCD nitric oxide (NO) production through activation of the ETB receptor during high-salt feeding. We determined that after 7 days of a high-salt diet (HS7), there was a shift to 100% ETB expression in IMCDs, as well as a twofold increase in nitrite production (a metabolite of NO), and this increase could be prevented by acute inhibition of the ETB receptor. ETB receptor blockade or NOS1 inhibition also prevented the ET-1-dependent decrease in ion transport from primary IMCDs, as determined by transepithelial resistance. IMCD were also isolated from vascular endothelial ET-1 knockout mice (VEETKO), collecting duct ET-1 KO (CDET-1KO), and flox controls. Nitrite production by IMCD from VEETKO and flox mice was similarly increased twofold with HS7. However, IMCD NO production from CDET-1KO mice was significantly blunted with HS7 compared with flox control. Taken together, these data indicate that during high-salt feeding, the autocrine actions of ET-1 via upregulation of the ETB receptor are critical for IMCD NO production, facilitating inhibition of ion reabsorption.

Regulation of metalloproteinases by nitric oxide in human trophoblast cells in culture

2001 ◽  
Vol 13 (6) ◽  
pp. 411 ◽  
Author(s):  
Virginia Novaro ◽  
Alejandro Colman-Lerner ◽  
Felipe Vadillo Ortega ◽  
Alicia Jawerbaum ◽  
Dante Paz ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Embryo Implantation ◽  
No Production ◽  
Trophoblast Cells ◽  
Human Trophoblast ◽  
No Donors ◽  
Multinucleated Cells ◽  
Nos Inhibitors ◽  
Spermine Nonoate

The process of embryo implantation requires extensive remodelling of the endometrial extracellular matrix, a function largely performed by matrix-degrading metalloproteinases (MMPs). In the present study, we used trophoblast cells isolated from human term placentas to study the regulation of MMPs by nitric oxide (NO). Using a combination of zymography, Western blot and indirect immunofluorescence, we showed that MMP-2 and MMP-9 are increased during the conversion from low-motile cytotrophoblast cells to the highly motile and differentiated syncytiotrophoblast multinucleated cells. We also observed an increase in NO production and NO synthase (NOS) expression during this cellular differentiation process. In addition, we demonstrated a positive regulatory role of NO on the activity and protein expression of MMP-2 and MMP-9, because NO donors (NOC-18 and spermine-NONOate) or the NOS substrate (L-arginine) stimulate, whereas NOS inhibitors (NG-nitro-L-arginine methyl ester and NG-monomethyl-L-arginine) reduce the expression and gelatinolytic activity of MMP-2 and MMP-9 in isolated trophoblast cells. Taken together, these results suggest that, in differentiating trophoblasts, NO regulates the induction of matrix-degrading proteases required for invasion during embryo implantation.

scholarly journals Flow stimulates inner medullary collecting duct endothelin‐1 production

2013 ◽  
Vol 27 (S1) ◽  
Author(s):  
Meghana Pandit ◽  
Gabriele L Gusella ◽  
Rajeev Rohatgi ◽  
Donald E Kohan
Keyword(s):  
Collecting Duct ◽  
Endothelin 1 ◽  
Inner Medullary Collecting Duct ◽  
Medullary Collecting Duct

Vasopressin-induced nitric oxide production in rat inner medullary collecting duct is dependent on V2 receptor activation of the phosphoinositide pathway

2007 ◽  
Vol 293 (2) ◽  
pp. F526-F532 ◽  
Author(s):  
Paul M. O'Connor ◽  
Allen W. Cowley
Keyword(s):  
Nitric Oxide ◽  
Adenylyl Cyclase ◽  
Collecting Duct ◽  
Receptor Activation ◽  
No Production ◽  
Moderate Increase ◽  
Inner Medullary Collecting Duct ◽  
Intracellular Ca ◽  
Medullary Collecting Duct ◽  
Phosphoinositide Pathway

We previously reported that arginine vasopressin (AVP) stimulates the production of nitric oxide (NO) in inner medullary collecting duct (IMCD) via activation of V2 receptors (V2R) and the mobilization of intracellular Ca2+. The aim of this study was to determine the pathway(s) through which this response is mediated. IMCDs were dissected from male Sprague-Dawley rats and intracellular Ca2+ concentration ([Ca2+]i) and NO production were measured using a fluorescence imaging system. AVP (100 nmol/l) produced a rapid increase [Ca2+]i of 381 ± 78 nmol/l that was followed by a significant increase of NO production (166 ± 61%). The specific nonpeptide V2R antagonist OPC31260 (1 μM), but not the V1R antagonist OPC21268 (1 μM), inhibited the increase in [Ca2+]i (up to 91 ± 5%) and abolished the NO response to AVP. Both the phospholipase C inhibitor U73112 (3 μM) and the inositol ( 1 , 4 , 5 ) tri-phosphate 3 receptor blocker 2-APB (75 μM) reduced the peak [Ca2+]i response to AVP (by 65 ± 9 and 59 ± 15%, respectively) and abolished the NO response. Although forskolin (100 μM; an activator of adenylyl cyclase) elicited a moderate increase in [Ca2+]i, neither preincubation with the adenylyl cyclase inhibitor 2′-5′-dideoxyadenosine (50 μM) nor the protein kinase A (PKA) inhibitor PKA14-22 (100 μM) significantly inhibited peak [Ca2+]i in response to AVP. IMCD [Ca2+]i responses to AVP were reduced by 72 ± 8% when incubated in Ca2+-free media and could be completely abolished by preincubation with the Ca2+-ATPase inhibitor thapsigargin. We conclude that AVP-induced NO production in IMCD is dependent on V2R activation of the phosphoinositide pathway and the mobilization of Ca2+ from both intracellular and extracellular pools.

The beta 1- and beta 2-adrenoceptor subtypes in cultured rat inner medullary collecting duct cells

1996 ◽  
Vol 271 (3) ◽  
pp. F762-F769 ◽  
Author(s):  
G. Yasuda ◽  
L. Sun ◽  
H. C. Lee ◽  
S. Umemura ◽  
W. B. Jeffries
Keyword(s):  
Radioligand Binding ◽  
Binding Capacity ◽  
Collecting Duct ◽  
Camp Accumulation ◽  
Single Class ◽  
Inner Medullary Collecting Duct ◽  
Beta Adrenoceptor ◽  
Site Model ◽  
Medullary Collecting Duct ◽  
Beta 2

We investigated beta-adrenoceptor subtype(s) expressed in cultured rat inner medullary collecting duct (IMCD) cells. In radioligand binding assay, [125I]iodocyanopindolol bound to IMCD cell membranes, representing a single class of binding sites (dissociation constant = 96.1 pM, maximum binding capacity = 18.2 fmol/mg protein, n = 8). In competition studies, ICI-89406 (beta 1-antagonist) and ICI-118551 (beta 2-antagonist) bound with high affinity, fitting a two-site model. Isoproterenol increased intracellular adenosine 3',5'-cyclic monophosphate (cAMP) accumulation (half-maximal effective concentration = 200 nM). Propranolol completely inhibited isoproterenol-induced cAMP accumulation [half-maximal inhibitory concentration (IC50) = 270 nM]. ICI-89406 and ICI-118551 inhibited cAMP accumulation by 50% (IC50 = 1.5 microM and 1.7 microM, respectively). The combined addition of ICI-89406 and ICI-118551 resulted in a curve indistinguishable from that of propranolol. The beta 1- and beta 2-adrenoceptor mRNAs have been demonstrated using reverse transcription-polymerase chain reaction. In initial and terminal IMCD cells, propranolol (3 microM) inhibited isoproterenol-stimulated cAMP accumulation by 80%, whereas ICI-89406 (3 microM) and ICI-118551 (3 microM) resulted in only partial inhibition (50%). We conclude that both beta 1- and beta 2-adrenoceptors are expressed in initial and terminal IMCD cells in primary culture.

Endothelin-1 production by rat inner medullary collecting duct: effect of nitric oxide, cGMP, and immune cytokines

1994 ◽  
Vol 266 (2) ◽  
pp. F291-F297 ◽  
Author(s):  
D. E. Kohan ◽  
E. Padilla
Keyword(s):  
Nitric Oxide ◽  
Collecting Duct ◽  
Sodium Reabsorption ◽  
Mrna Levels ◽  
Endothelin 1 ◽  
Inner Medullary Collecting Duct ◽  
Short And Long Term ◽  
Medullary Collecting Duct ◽  
Necrosis Factor

Nitric oxide (NO), guanosine 3',5'-cyclic monophosphate (cGMP), and endothelin-1 (ET-1) inhibit collecting duct sodium reabsorption. Because the inner medullary collecting duct (IMCD) synthesizes NO and ET-1, we examined NO and cGMP regulation of IMCD ET-1 production. S-nitroso-N-acetylpenicillamine (SNAP, 6 h) increased NO and cGMP and modestly reduced ET-1 release in cultured rat IMCD. Atrial natriuretic peptide or dibutyryl cGMP (6 h exposure to each) also mildly decreased IMCD ET-1 release. In long-term exposure studies, IMCD cells were incubated with tumor necrosis factor (TNF) and interferon-gamma (IFN) up to 72 h. IFN/TNF increased NO and cGMP production while reducing ET-1 release by 84%; N-monomethyl-L-arginine inhibited this effect only marginally, suggesting NO was not primarily involved. IFN alone greatly reduced IMCD ET-1 release and ET-1 mRNA levels. These data indicate that short- and long-term increases in NO and cGMP modestly reduce IMCD ET-1 production. Additionally, IFN potently inhibits IMCD ET-1 release by an undetermined mechanism.

Shear stress-mediated NO production in inner medullary collecting duct cells

2000 ◽  
Vol 279 (2) ◽  
pp. F270-F274 ◽  
Author(s):  
Zheqing Cai ◽  
Jingdong Xin ◽  
David M. Pollock ◽  
Jennifer S. Pollock
Keyword(s):  
Shear Stress ◽  
Fluid Shear Stress ◽  
Collecting Duct ◽  
No Synthase ◽  
No Production ◽  
Fluid Shear ◽  
Water Excretion ◽  
Inner Medullary Collecting Duct ◽  
Nitrite Production ◽  
Medullary Collecting Duct

Recent evidence suggests that nitric oxide (NO) within the inner medullary collecting duct (IMCD) functions to regulate sodium and water reabsorption. Because fluid shear stress has been shown to increase NO production in endothelial and vascular smooth muscle cells, experiments were designed to determine whether a similar mechanism exists in IMCD cells. Cultured IMCD-3 cells derived from murine IMCD were subjected to 60 min of pulsatile shear stress. Nitrite production (2,3-diaminonaphthalene fluorometric assay) increased 12-, 16-, and 23-fold at 3.3, 10, and 30 dyn/cm2, respectively, compared with static control cultures. Preincubation with the non-isoform-specific NO synthase inhibitor nitro-l-arginine methyl ester reduced nitrite production by 83% in response to 30 dyn/cm2. Western blotting and immunofluorescence analysis of static IMCD-3 cell cultures revealed the expression of all three NO synthase isoforms (NOS-1 or neuronal NOS, NOS-2 or inducible NOS, and NOS-3 or endothelial NOS) in IMCD-3 cultures. These results indicate that NO production is modulated by shear stress in IMCD-3 cells and that fluid shear stress within the renal tubular system may play a role in the regulation of sodium and water excretion by control of NO production in the IMCD.

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