Regulation of transepithelial ion transport by two different purinoceptors in the apical membrane of canine kidney (MDCK) cells

Zegarra‐Moran, Olga; Romeo, Giovanni; Galietta, Luis J. V.

doi:10.1111/j.1476-5381.1995.tb13312.x

Cited by 51 publications

(33 citation statements)

References 21 publications

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“…P2Y 12 , P2Y 13 ) are known to utilize G i -dependent mechanisms to inhibit cAMP formation (29,30), whereas other P2Y receptors may regulate adenylyl cyclase activity via effects on regulators of adenylyl cyclase, such as [Ca 2ϩ ] or protein kinase C. MDCK-D 1 cells, in common with several other cell types, express several different P2Y receptor subtypes (1,6,12,31,32). Early work on MDCK-D 1 cells, prior to the molecular cloning and precise identification of the different P2Y receptor subtypes, emphasized the ability of added nucleotides to alter membrane potential, ion flux, or short circuit current (7,8). Recent studies have indicated that MDCK-D 1 cells release ATP and that this released nucleotide helps contribute to basal activity of signal transduction pathways (15,33).…”

Section: Resultsmentioning

confidence: 99%

“…5 and 6). P2Y receptors in MDCK-D 1 cells modulate membrane potential and short circuit current, and the receptors regulate phospholipases, intracellular [Ca 2ϩ ], prostaglandin E 2 (PGE 2 ) formation, and cAMP accumulation (1,(7)(8)(9)(10)(11)(12)(13)(14).Cyclooxygenase (COX) inhibitors, such as indomethacin (indo), have proven useful for studying P2Y receptors in MDCK-D 1 cells (1, 12). Agonist-stimulated cAMP accumulation in MDCK-D 1 cells occurs via both indo-sensitive and -insensitive pathways.…”

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confidence: 99%

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P2Y11 Receptors Activate Adenylyl Cyclase and Contribute to Nucleotide-promoted cAMP Formation in MDCK-D1Cells

Torres

Zambon

Insel

2002

Journal of Biological Chemistry

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Cells commonly co-express multiple receptor subtypes that recognize the same physiological agonist, but it is difficult to define which among such receptor subtypes mediates a particular response. This can be a particularly vexing problem if subtype-selective agonists and antagonists are not available. One such example is P2Y receptors, which respond to ATP and other nucleotides, are expressed in a variety of tissues and cell types, and for which few subtype-selective antagonists exist (2-4). Our laboratory has undertaken a series of studies related to signal transduction by P2Y receptors in the renal epithelial cell line, MDCK 1 -D 1 (reviewed in Refs. 5 and 6). P2Y receptors in MDCK-D 1 cells modulate membrane potential and short circuit current, and the receptors regulate phospholipases, intracellular [Ca 2ϩ ], prostaglandin E 2 (PGE 2 ) formation, and cAMP accumulation (1,(7)(8)(9)(10)(11)(12)(13)(14).Cyclooxygenase (COX) inhibitors, such as indomethacin (indo), have proven useful for studying P2Y receptors in MDCK-D 1 cells (1, 12). Agonist-stimulated cAMP accumulation in MDCK-D 1 cells occurs via both indo-sensitive and -insensitive pathways. Response to the P2Y 2 agonist UTP is entirely indo-sensitive, whereas response to ATP is partially sensitive and to 2-methylthio-ATP (MT-ATP) is insensitive (1). These findings suggest that UTP, and ATP in part, stimulate P2Y 2 receptors to cause COX-mediated (perhaps by both COX1 and COX2, see Ref. 15) formation of arachidonic acid metabolites (e.g. PGE 2 ), which activate EP receptors to stimulate cAMP formation, while ATP and MT-ATP can also enhance cAMP formation via an indo-insensitive P2Y receptor pathway (1).The present studies were designed to characterize more fully the nature of the latter pathway. Our working hypothesis, based in part on initial results obtained with MDCK-D 1 cells (12), was that the indo-resistant response might represent a P2Y 1 receptor effect. The current data show results not consistent with this hypothesis but instead suggest a key role for another receptor, the P2Y 11 receptor, in indo-resistant cAMP formation. The findings directly document a role for P2Y 11 receptors in stimulation of adenylyl cyclase activity and in potentially contributing to autocrine-paracrine regulation by nucleotides. EXPERIMENTAL PROCEDURESCell Culture-MDCK-D 1 cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% mixed serum (85% horse serum, 15% fetal bovine serum). Cells were used in assays at 60 -80% confluency. GFP-tagged cP2Y 11 receptor-overexpressing MDCK-D 1 cells were cultured from the stable cell line prepared by Zambon et al. (14).Measurement of cAMP Accumulation in Normal and GFP-tagged cP2Y 11 Intact Cells-Prior to the treatment of the cells, the growth medium was removed, and cells were equilibrated for 30 min at 37°C in serum-free 20 mM HEPES-buffered Dulbecco's modified Eagle's medium (DMEH, pH 7.4). Subsequently, cells were incubated in fresh DMEH and various agents as shown in the figures. Incubations with th...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

P2Y11 Receptors Activate Adenylyl Cyclase and Contribute to Nucleotide-promoted cAMP Formation in MDCK-D1Cells

Torres

Zambon

Insel

2002

Journal of Biological Chemistry

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“…A later paper from this group showed that ATP increased [Ca 2+ ] i ; calcium then activates K + channels and thus leads to hyperpolarisation of the cell membrane [281]. Regulation of transepithelial ion transport by two different receptors on the apical membrane of MDCK cells was claimed [430], the data implicating P2Y 1 and P2Y 2 (or P2Y 4 ) receptors. Lanthanum inhibits UTP-induced Ca 2+ mobilization in MDCK cells [164].…”

Section: Distal Tubulesmentioning

confidence: 99%

Purinergic signalling in the kidney in health and disease

Burnstock

Evans

Bailey

2013

Purinergic Signalling

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The involvement of purinergic signalling in kidney physiology and pathophysiology is rapidly gaining recognition and this is a comprehensive review of early and recent publications in the field. Purinergic signalling involvement is described in several important intrarenal regulatory mechanisms, including tuboglomerular feedback, the autoregulatory response of the glomerular and extraglomerular microcirculation and the control of renin release. Furthermore, purinergic signalling influences water and electrolyte transport in all segments of the renal tubule. Reports about purine-and pyrimidine-mediated actions in diseases of the kidney, including polycystic kidney disease, nephritis, diabetes, hypertension and nephrotoxicant injury are covered and possible purinergic therapeutic strategies discussed.

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“…5 The intact renal tubule and MDCK and other renal epithelial cells are known to express numerous luminal and basolateral P2 receptors. 6,11 Their activation commonly triggers an increase of [Ca 2ϩ ] i . 12 Therefore, we hypothesized that flow-induced [Ca 2ϩ ] i signals in renal epithelia involve release of nucleotides and associated signaling events.…”

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confidence: 99%

Flow-Induced [Ca2+]i Increase Depends on Nucleotide Release and Subsequent Purinergic Signaling in the Intact Nephron

Jensen

Odgaard

Christensen

et al. 2007

Journal of the American Society of Nephrology

107

158

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Flow induces cytosolic Ca 2ϩ increases ([Ca 2ϩ ] i ) in intact renal tubules, but the mechanism is elusive. Mechanical stimulation in general is known to promote release of nucleotides (ATP/UTP) and trigger auto-and paracrine activation of P2 receptors in renal epithelia. It was hypothesized that the flowinduced [Ca 2ϩ ] i response in the renal tubule involves mechanically stimulated nucleotide release. This study investigated (1) the expression of P2 receptors in mouse medullary thick ascending limb (mTAL) using P2Y 2 receptor knockout (KO) mice, (2) whether flow increases induce [Ca 2ϩ ] i elevations in mTAL, and (3) whether this flow response is affected in mice that are deplete of the main purinergic receptor. [Ca 2ϩ ] i was imaged in perfused mTAL with fura-2 or fluo-4. It is shown that luminal and basolateral P2Y 2 receptors are the main purinergic receptor in this segment. Moreover, the data suggest presence of basolateral P2X receptors. Increases of tubular flow were imposed by promptly rising the inflow pressure, which triggered a marked increase of [Ca 2ϩ ] i . This [Ca 2ϩ ] i response was significantly reduced in P2Y 2 receptor KO tubules (fura-2 ratio increase WT 0.44 Ϯ 0.09 [n ϭ 28] versus KO 0.16 Ϯ 0.04 [n ϭ 13]). Furthermore, the flow response was greatly inhibited with luminal and basolateral scavenging of extracellular ATP (apyrase 7.5 U/ml) or blockage of P2 receptors (suramin 300 M). The flow response could still be elicited in the absence of extracellular Ca 2ϩ . These results strongly suggest that increase of tubular flow elevates [Ca 2ϩ ] i in intact renal epithelia. This flow response is caused by release of bilateral nucleotides and subsequent activation of P2 receptors.

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Regulation of transepithelial ion transport by two different purinoceptors in the apical membrane of canine kidney (MDCK) cells

Cited by 51 publications

References 21 publications

P2Y11 Receptors Activate Adenylyl Cyclase and Contribute to Nucleotide-promoted cAMP Formation in MDCK-D1Cells

P2Y11 Receptors Activate Adenylyl Cyclase and Contribute to Nucleotide-promoted cAMP Formation in MDCK-D1Cells

Purinergic signalling in the kidney in health and disease

Flow-Induced [Ca2+]i Increase Depends on Nucleotide Release and Subsequent Purinergic Signaling in the Intact Nephron

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