Prostaglandin-concentration-dependent desensitization of adenylate cyclase in human erythroleukaemia (HEL) cells is abolished by pertussis toxin and enhanced by induction by dimethyl sulphoxide

Ashby, Barrie; Almonor, G O; Wernick, Eva; Selak, Mary

doi:10.1042/bj2800801

Cited by 11 publications

(8 citation statements)

References 17 publications

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“…Differentiation with dimethylsulphoxide activates protein kinase C in murine erythroleukaemia cells (Chakraverthy et al, 1992) and accelerates the desensitization rate of G,-coupled prostaglandin E receptors in human erythroleukaemia cells (HEL-cells; Ashby et al, 1991). This differentiation also lowered peak adrenaline-or NPY-stimulated Ca2l elevations, which is compatible with the idea of enhanced desensitization rates for these Gi-mediated responses.…”

Section: Discussionsupporting

confidence: 68%

Rapid desensitization of adrenaline‐ and neuropeptide Y‐stimulated Ca²⁺ mobilization in HEL‐cells

Michel

1994

British J Pharmacology

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1 Desensitization of G,-coupled receptors, the P2-adrenoceptor for example, involves rapid and slower components but little is known regarding the existence of rapid desensitization of Gi-coupled 2 Following stimulation by adrenaline or neuropeptide Y Ca2+ levels returned towards baseline a few minutes after agonist addition and were refractory to a second agonist exposure demonstrating rapid desensitization. Cross-desensitization experiments with neuropeptide Y, adrenaline and moxonidine demonstrated the presence of homologous (both receptors) and heterologous desensitization (neuropeptide Y receptors only), and that the x2A-adrenoceptor desensitization was not specific for phenylethylamine (adrenaline) or imidazoline agonists (moxonidine). 3 The protein kinase C activator, phorbol ester, rapidly desensitized the hormonal Ca2+ responses and inhibitors of protein kinase C enhanced the hormonal responses inconsistently. The tyrosine kinase inhibitor, herbimycin, enhanced Ca2+ mobilization by adrenaline and neuropeptide Y, whereas the protein phosphatase inhibitor, okdadaic acid, did not affect Ca2+ mobilization or its desensitization. 4 In the absence of extracellular Ca2+ the endoplasmic reticulum Ca2+-ATPase inhibitor, thapsigargin, reduced hormone-stimulated Ca2+ elevations, demonstrating that mobilization occurs from a thapsigargin-sensitive pool in the endoplasmic reticulum. The inositol phosphate-independent Ca2' release modulator, ryanodine, significantly enhanced adrenaline-and neuropeptide Y-stimulated Ca2+ elevations. Blockade of the endoplasmic reticulum Ca2+-ATPase by thapsigargin in the presence of extracellular Ca2+ enhanced hormone-stimulated Ca2+ increases, demonstrating the importance of this enzyme for the termination of the Ca2+ signal. 5 It is concluded that adrenaline and neuropeptide Y-stimulated Ca2+ mobilization in HEL-cells occurs from a thapsigargin-and ryanodine-sensitive store in the endoplasmic reticulum and desensitizes rapidly; this appears to involve multiple mechanisms including protein kinases, possibly acting on receptors, and Ca2+ release and sequestration mechanisms.

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Section: Discussionsupporting

confidence: 68%

Rapid desensitization of adrenaline‐ and neuropeptide Y‐stimulated Ca²⁺ mobilization in HEL‐cells

Michel

1994

British J Pharmacology

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“…However, several different agonists that stimulate adenylyl cyclase via Gscoupled receptors have been studied in HEL cells and occasionally in other human erythroleukaemia cell lines (Ashby et al 1991;M~ki et al 1992;Feoktistov and Biaggioni 1993). Moreover, the presence of the fl-adrenoceptorcyclase system has been documented repeatedly in erythroid cells of other mammalian species (Beckman et al 1980;Setchenska et al 1983;Sytkowski and Kessler 1984;Moudry and Porzig 1990).…”

Section: Receptor-mediated Effects On Adenylyl Cyclasementioning

confidence: 99%

G-protein-coupled receptors in normal human erythroid progenitor cells

Porzig

Gutknecht

Kostova

et al. 1995

Naunyn-Schmiedeberg's Arch Pharmacol

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Human erythroid progenitor cells were isolated from peripheral blood of healthy donors and amplified in a suspension culture system using recombinant growth factors (stem cell factor, interleukin-3, granulocyte-macrophage colony-stimulating factor and erythropoietin) as well as conditioned medium from a human bone marrow stroma cell line to support cell proliferation. After 6-8 days of culture, the cell population consisted mainly of erythroid colony-forming cells (burst-forming units, BFU-Es and colony-forming units, CFU-Es). In these cells, we studied ligand-induced changes in intracellular Ca2+ concentration ([Ca2+]i) and cAMP formation as the primary effector systems of guanine nucleotide-binding protein (G protein)-coupled receptors. The results confirmed the functional expression of receptors for adenosine (type A2B), prostaglandin E1 and isoprenaline (beta-adrenoceptor), all of which stimulated adenylyl cyclase, as well as for ADP (purinoceptor types P2T and P2U), platelet-activating factor and thrombin all of which caused a transient increase in [Ca2+]i. The efficacy of adenosine and prostaglandin E1 in stimulating cAMP formation was more than 5 times higher than that of isoprenaline, suggesting a low beta-adrenoceptor density. The response to adenosine and isoprenaline decreased by 80 and 55% respectively during maturation into the proerythroblast stage. Similarly, thapsigargin-sensitive intracellular Ca2+ stores and ligand-induced Ca2+ release declined by about 60% during the CFU-E-to-erythroblast transition. The overall functional expression pattern of G protein-coupled receptors differed from that in human erythroleukaemia cell lines or from that in platelets. Primary culture systems for nontransformed cells, such as the one presented here, thus will be indispensable for the study of the functional role of G protein-dependent signalling during haematopoiesis.

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“…Sequencing reagents were from United States culture flasks in an atmosphere of 95 % air/5 % CO2 at 37 'C. Cells were grown for 4 days in the presence of 1.25 % dimethyl sulphoxide (DMSO) to induce prostaglandin receptors [3]. Sequence comparisons and translations were carried out using the GCG Sequence Analysis Package (University of Wisconsin).…”

Section: Kb Longmentioning

confidence: 99%

“…We have suggested that prostaglandin regulation of cyclic AMP metabolism in platelets involves separate stimulatory and inhibitory prostaglandin receptors linked to adenylate cyclase, giving rise to characteristic patterns of cyclic AMP formation that may buffer the cellular response to localized changes in prostaglandin concentration [1,2]. We have shown similar regulation of cyclic AMP metabolism in human erythroleukaemia (HEL) cells [3], a megakaryocytic cell line possessing several platelet-like features [4] that has been used for cloning of platelet-specific receptors such as the thrombin receptor [5].…”

Section: Introductionmentioning

confidence: 98%

Cloning and expression of a prostaglandin E receptor EP3 subtype from human erythroleukaemia cells

Kunapuli

Mao

Baştepe

et al. 1994

Biochemical Journal

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Prostaglandins inhibit platelet activation by stimulating intracellular cyclic AMP formation. We have postulated that intracellular cyclic AMP levels in platelets are buffered by a distinct prostaglandin receptor that mediates inhibition of cyclic AMP formation. In order to provide evidence for the model, we have cloned the cDNA coding for a prostaglandin receptor EP3 subtype, which is coupled to inhibition of adenylate cyclase, from the megakaryocytic cell line human erythroleukaemia (HEL) cells. A PCR-generated hybridization probe, produced using primers based on the sequence of the mouse prostaglandin EP3 receptor published by Sugimoto, Namba, Honda, Hayashi, Negishi, Ichikawa and Narumiya [(1992) J. Biol. Chem. 267, 6463-6466], was used to screen a lambda gt11 HEL cell cDNA library. The composite full-length cDNA clone HEP3, generated from the two partial clones pHEP3-7 and pHEP3-5, is 1.6 kb long with an open reading frame coding for 390 amino acids. This clone is 83% identical to the alpha subtype of the mouse EP3 receptor. The full-length construct was transfected into COS-1 cells. The cloned receptor exhibited the properties of a prostaglandin EP3 subtype, inhibiting forskolin-stimulated cyclic AMP formation in response to prostaglandin E2 (PGE2) and binding PGE2 with high specificity and a Kd of 3.2 nM. Radiolabelled PGE2 could be displaced by prostaglandins in the order PGE2 = PGE1 > iloprost = PGD2. Northern blot analysis revealed that the receptor is also present in human kidney.

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Prostaglandin-concentration-dependent desensitization of adenylate cyclase in human erythroleukaemia (HEL) cells is abolished by pertussis toxin and enhanced by induction by dimethyl sulphoxide

Cited by 11 publications

References 17 publications

Rapid desensitization of adrenaline‐ and neuropeptide Y‐stimulated Ca²⁺ mobilization in HEL‐cells

Rapid desensitization of adrenaline‐ and neuropeptide Y‐stimulated Ca²⁺ mobilization in HEL‐cells

G-protein-coupled receptors in normal human erythroid progenitor cells

Cloning and expression of a prostaglandin E receptor EP3 subtype from human erythroleukaemia cells

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Prostaglandin-concentration-dependent desensitization of adenylate cyclase in human erythroleukaemia (HEL) cells is abolished by pertussis toxin and enhanced by induction by dimethyl sulphoxide

Cited by 11 publications

References 17 publications

Rapid desensitization of adrenaline‐ and neuropeptide Y‐stimulated Ca2+ mobilization in HEL‐cells

Rapid desensitization of adrenaline‐ and neuropeptide Y‐stimulated Ca2+ mobilization in HEL‐cells

G-protein-coupled receptors in normal human erythroid progenitor cells

Cloning and expression of a prostaglandin E receptor EP3 subtype from human erythroleukaemia cells

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Rapid desensitization of adrenaline‐ and neuropeptide Y‐stimulated Ca²⁺ mobilization in HEL‐cells

Rapid desensitization of adrenaline‐ and neuropeptide Y‐stimulated Ca²⁺ mobilization in HEL‐cells