Basolateral Translocation by Vasopressin of the Aldosterone-Induced Pool of Latent Na-K-ATPases Is Accompanied by α1 Subunit Dephosphorylation: Study in a New Aldosterone-Sensitive Rat Cortical Collecting Duct Cell Line

Abstract. The regulation of plasma membrane Na+-K+-ATPases (NKA) expression by aldosterone and arginin vasopressin (AVP) in the cortical collecting duct (CCD) has been examined in a new rat CCD cell line, designated as RCCD2. This cell line has maintained many characteristics of the CCD—in particular, the expression of the mineralocorticoid receptor. Mineralocorticoid receptor is expressed at the protein level and binds3H-aldosterone (approximately 15 to 20 fmol/mg protein). Short-circuit current (Isc) experiments showed approximately a twofold increase in Isc associated with a decrease in transepithelial resistance when cells were treated with aldosterone concentrations as low as 10-9M. This effect on Isc was significant 2 h after aldosterone addition and was still present after 24 h. It was accompanied by an increase in the amount of mRNA encoding for the α subunit of the epithelial sodium channel (sixfold) and the α1 subunit of NKA (fourfold) after 24 h of hormone treatment. In addition, mRNA expression of the serum- and glucocorticoid-induced kinase (Sgk) was increased by 10-9M aldosterone treatment as early as 45 min after hormone addition. As had already been documented in native CCD obtained by microdissection, incubation of RCCD2cells for 24 h with aldosterone resulted in the constitution of a latent pool of NKA that could be rapidly recruited by AVP (15 min). NKA biotinylation experiments and preparation of membrane fractions show that this latent pool of NKA is present in the intracellular compartment of the cells and is recruited by AVP in the basolateral membrane through a translocation process. This mechanism is accompanied by dephosphorylation of the α1catalytic subunit of NKA.

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Activation of the Amiloride-Sensitive Epithelial Sodium Channel by the Serine Protease mCAP1 Expressed in a Mouse Cortical Collecting Duct Cell Line

Journal of the American Society of Nephrology ◽

10.1681/asn.v115828 ◽

2000 ◽

Vol 11 (5) ◽

pp. 828-834

Author(s):

GRÉGOIRE VUAGNIAUX ◽

VÉRONIQUE VALLET ◽

NICOLE FOWLER JAEGER ◽

CORINNE PFISTER ◽

MARCELLE BENS ◽

...

Keyword(s):

Sodium Channel ◽

Cell Line ◽

Serine Protease ◽

Sodium Transport ◽

Serine Proteases ◽

Collecting Duct ◽

Short Circuit ◽

Short Circuit Current ◽

Duct Cell ◽

Cortical Collecting Duct

Abstract. This study examines whether serine proteases can activate the amiloride-sensitive sodium channel (ENaC) in mammalian kidney epithelial cells. The transepithelial sodium transport assessed by amiloride-sensitive short-circuit current appears to be sensitive to aprotinin, a protease inhibitor in a mouse cortical collecting duct cell line (mpkCCDc14). This result indicated that serine proteases may be implicated in the regulation of ENaC-mediated sodium transport. Using degenerated oligonucleotides to a previously isolated serine protease from Xenopus, xCAP1 (channel activating protease), a novel full-length serine protease (mCAP1), has been isolated and characterized. RNA analysis showed a broad pattern of expression in tissues (kidney, lung, colon, and salivary glands) expressing ENaC. Reverse transcription-PCR experiments also showed that mCAP1 was abundantly expressed in proximal tubule cells and was also expressed in intact and cultured collecting duct cells. Coexpression of the Xenopus, rat, or human α-, β-, and γ-ENaC subunits in Xenopus oocytes also showed that mCAP1 induces a significant increase in ENaC-mediated current accompanied by a decrease of channel molecules at the cell surface. It is proposed that this novel mouse channel activating protease may act as a regulator of ENaC within the kidney.

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ANF and bradykinin synergistically inhibit transport in M-1 cortical collecting duct cell line

AJP Renal Physiology ◽

10.1152/ajprenal.1992.263.1.f1 ◽

1992 ◽

Vol 263 (1) ◽

pp. F1-F6 ◽

Cited By ~ 3

Author(s):

B. A. Stoos ◽

O. A. Carretero ◽

J. L. Garvin

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Cell Line ◽

Collecting Duct ◽

Short Circuit ◽

Short Circuit Current ◽

Duct Cell ◽

Cortical Collecting Duct ◽

Water Excretion ◽

Kinase C

Previous data suggest that atrial natriuretic factor (ANF) and bradykinin (BK) interact to increase Na+ and water excretion. We propose that this interaction is due to a synergistic action that inhibits Na+ absorption in the distal nephron. We examined the effects of BK and ANF on transport by monolayers of a cortical collecting duct cell line, M-1. BK (10(-8) M) had no effect on short-circuit current (Isc). Similarly, ANF (10(-8) M) did not inhibit Isc. In contrast, Isc decreased by 18% (from 57 +/- 8 to 46 +/- 6 microA/cm2) when BK and ANF were added simultaneously at this concentration (P less than 0.05). Because guanosine 3',5'-cyclic monophosphate (cGMP) and protein kinase C are implicated in the second messenger cascades of ANF and BK, we investigated their potential roles in mediating this interaction. Dibutyryl-cGMP (10(-4) M) inhibited Isc from 33 +/- 4 to 22 +/- 3 microA/cm2 (P less than 0.05) in the presence of BK but not in its absence. Staurosporine and calphostin C, inhibitors of protein kinase C, completely blocked the decrease in Isccaused by simultaneous addition of ANF and BK. cAMP levels in M-1 cells were not affected by either ANF alone or BK alone; however, when cultures were treated with both hormones, cAMP decreased from 856 +/- 56 to 332 +/- 26 fmol/10(6) cells (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

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Site-1 Protease-Derived Soluble (Pro)Renin Receptor Contributes to Angiotensin II–Induced Hypertension in Mice

Hypertension ◽

10.1161/hypertensionaha.120.15100 ◽

2020 ◽

Author(s):

Ye Feng ◽

Kexin Peng ◽

Renfei Luo ◽

Fei Wang ◽

Tianxin Yang

Keyword(s):

Blood Pressure ◽

Angiotensin Ii ◽

Collecting Duct ◽

Short Circuit ◽

Ussing Chamber ◽

Short Circuit Current ◽

Duct Cell ◽

Ang Ii ◽

Cortical Collecting Duct ◽

Induced Hypertension

Activation of PRR ([pro]renin receptor) contributes to enhancement of intrarenal RAS and renal medullary α-ENaC and thus elevated blood pressure during Ang II (angiotensin II) infusion. The goal of the present study was to test whether such action of PRR was mediated by sPRR (soluble PRR), generated by S1P (site-1 protease), a newly identified PRR cleavage protease. F1 B6129SF1/J mice were infused for 6 days with control or Ang II at 300 ng/kg per day alone or in combination with S1P inhibitor PF-429242 (PF), and blood pressure was monitored by radiotelemetry. S1P inhibition significantly attenuated Ang II–induced hypertension accompanied with suppressed urinary and renal medullary renin levels and expression of renal medullary but not renal cortical α-ENaC expression. The effects of S1P inhibition were all reversed by supplement with histidine-tagged sPRR termed as sPRR-His. Ussing chamber technique was performed to determine amiloride-sensitive short-circuit current, an index of ENaC activity in confluent mouse cortical collecting duct cell line cells exposed for 24 hours to Ang II, Ang II + PF, or Ang II + PF + sPRR-His. Ang II–induced ENaC activity was blocked by PF, which was reversed by sPRR-His. Together, these results support that S1P-derived sPRR mediates Ang II–induced hypertension through enhancement of intrarenal renin level and activation of ENaC.

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The isolated C-terminus of polycystin-1 promotes increased ATP-stimulated chloride secretion in a collecting duct cell line

Clinical Science ◽

10.1042/cs1040217 ◽

2003 ◽

Vol 104 (3) ◽

pp. 217-221 ◽

Cited By ~ 12

Author(s):

Kimberly M. HOOPER ◽

Robert J. UNWIN ◽

Michael SUTTERS

Keyword(s):

Fusion Protein ◽

Cell Line ◽

Collecting Duct ◽

Basolateral Membrane ◽

Duct Cell ◽

Chloride Secretion ◽

Cortical Collecting Duct ◽

Membrane Expression ◽

Fusion Protein Expression ◽

C Terminus

Cyst expansion in autosomal dominant polycystic kidney disease (ADPKD) requires accumulation of fluid into the cyst lumen, which is probably driven by aberrant chloride secretion by the cyst lining epithelium. Extracellular ATP is a potent stimulus for chloride secretion in many epithelial systems, and provides a plausible mechanism for secretion in ADPKD. Therefore the link between polycystin-1 and ATP-stimulated chloride secretion was investigated in the M1 cortical collecting duct cell line. M1 cells were stably transfected with a glucocorticoid-inducible cytoplasmic C-terminal polycystin-1 construct fused to a membrane expression cassette. Induction of fusion protein expression was associated with augmentation of ATP-stimulated transepithelial chloride secretion. After nystatin-induced permeabilization of the basolateral membrane, it was determined that expression of the polycystin fusion protein modulated an ATP-responsive apical chloride conductance. It is concluded that up-regulation of ATP-stimulated chloride secretion might play a significant role in cyst expansion in ADPKD.

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Osmotic Stress Regulates Mineralocorticoid Receptor Expression in a Novel Aldosterone-Sensitive Cortical Collecting Duct Cell Line

Molecular Endocrinology ◽

10.1210/me.2009-0095 ◽

2009 ◽

Vol 23 (12) ◽

pp. 1948-1962 ◽

Cited By ~ 33

Author(s):

Say Viengchareun ◽

Peter Kamenicky ◽

Marie Teixeira ◽

Daniel Butlen ◽

Geri Meduri ◽

...

Keyword(s):

Osmotic Stress ◽

Cell Line ◽

Mineralocorticoid Receptor ◽

Collecting Duct ◽

Duct Cell ◽

Receptor Expression ◽

Cortical Collecting Duct

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Actin-based cytoskeleton regulates a chloride channel and cell volume in a renal cortical collecting duct cell line.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)37249-6 ◽

1994 ◽

Vol 269 (10) ◽

pp. 7081-7089

Author(s):

E.M. Schwiebert ◽

J.W. Mills ◽

B.A. Stanton

Keyword(s):

Cell Volume ◽

Cell Line ◽

Chloride Channel ◽

Collecting Duct ◽

Duct Cell ◽

Cortical Collecting Duct

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Effects of chemical chaperones on partially retarded NaCl cotransporter mutants associated with Gitelman's syndrome in a mouse cortical collecting duct cell line

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfg474 ◽

2004 ◽

Vol 19 (5) ◽

pp. 1069-1076 ◽

Cited By ~ 10

Author(s):

J. C. de Jong ◽

P. H. G. M. Willems ◽

M. Goossens ◽

A. Vandewalle ◽

L. P. W. J. van den Heuvel ◽

...

Keyword(s):

Cell Line ◽

Collecting Duct ◽

Duct Cell ◽

Cortical Collecting Duct ◽

Chemical Chaperones ◽

Gitelman's Syndrome ◽

Nacl Cotransporter

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IGF-1 vs insulin: Respective roles in modulating sodium transport via the PI-3 kinase/Sgk1 pathway in a cortical collecting duct cell line

Kidney International ◽

10.1038/sj.ki.5002018 ◽

2007 ◽

Vol 71 (2) ◽

pp. 116-125 ◽

Cited By ~ 43

Author(s):

E. Gonzalez-Rodriguez ◽

H.-P. Gaeggeler ◽

B.C. Rossier

Keyword(s):

Cell Line ◽

Sodium Transport ◽

Collecting Duct ◽

Duct Cell ◽

Cortical Collecting Duct

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Modulation of aldosterone-induced stimulation of ENaC synthesis by changing the rate of apical Na+ entry

AJP Renal Physiology ◽

10.1152/ajprenal.2001.281.4.f687 ◽

2001 ◽

Vol 281 (4) ◽

pp. F687-F692 ◽

Cited By ~ 4

Author(s):

Lisette Dijkink ◽

Anita Hartog ◽

Carel H. Van Os ◽

René J. M. Bindels

Keyword(s):

Feedback Inhibition ◽

Collecting Duct ◽

Short Circuit ◽

Primary Cultures ◽

Short Circuit Current ◽

Inhibitory Effect ◽

Cortical Collecting Duct ◽

Entry Rate ◽

Protein Levels ◽

Stimulation Of

Primary cultures of immunodissected rabbit connecting tubule and cortical collecting duct cells were used to investigate the effect of apical Na+ entry rate on aldosterone-induced transepithelial Na+ transport, which was measured as benzamil-sensitive short-circuit current ( I sc). Stimulation of the apical Na+ entry, by long-term short-circuiting of the monolayers, suppressed the aldosterone-stimulated benzamil-sensitive I sc from 320 ± 49 to 117 ± 14%, whereas in the presence of benzamil this inhibitory effect was not observed (335 ± 74%). Immunoprecipitation of [35S]methionine-labeled β-rabbit epithelial Na+ channel (rbENaC) revealed that the effects of modulation of apical Na+ entry on transepithelial Na+ transport are exactly mirrored by β-rbENaC protein levels, because short-circuiting the monolayers decreased aldosterone-induced β-rbENaC protein synthesis from 310 ± 51 to 56 ± 17%. Exposure to benzamil doubled the β-rbENaC protein level to 281 ± 68% in control cells but had no significant effect on aldosterone-stimulated β-rbENaC levels (282 ± 68%). In conclusion, stimulation of apical Na+ entry suppresses the aldosterone-induced increase in transepithelial Na+transport. This negative-feedback inhibition is reflected in a decrease in β-rbENaC synthesis or in an increase in β-rbENaC degradation.

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