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PMID: 8052522 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Single-cell fura-2 microfluorometry reveals different purinoceptor subtypes coupled to Ca2+ influx and intracellular Ca2+ release in bovine adrenal chromaffin and endothelial cells.

Pflugers Archiv : European journal of physiology ·Vol. 426 ·No. 6 ·1994-04-00 ·Pages 524-33

Castro E, Tomé AR, Miras-Portugal MT, Rosário LM

Abstract

ATP and adenosine(5')tetraphospho(5')adenosine (Ap4A), released from adrenal chromaffin cells, are potent stimulators of endothelial cell function. Using single-cell fura-2 fluorescence recording techniques to measure free cytosolic Ca2+ concentration ([Ca2+]i), we have investigated the role of purinoceptor subtypes in the activation of cocultured chromaffin and endothelial cells. ATP evoked concentration-dependent [Ca2+]i rises (EC50 = 3.8 microM) in a subpopulation of chromaffin cells. Both ATP-sensitive and -insensitive cells were potently activated by nicotine, bradykinin and muscarine. Reducing extracellular free Ca2+ concentration to around 100 nM suppressed the [Ca2+]i transient evoked by ATP but not the [Ca2+]i response to bradykinin. ATP-sensitive chromaffin cells were also potently stimulated by 2-methylthioadenosine triphosphate (2MeSATP; EC50 = 12.5 microM) and UTP, but did not respond to either adenosine 5'-[beta-thio]diphosphate (ADP[beta S]), a P2Y receptor agonist, adenosine 5'-[alpha,beta-methylene]triphosphate (pp-[CH2]pA), a P2X agonist or AMP. Adrenal endothelial cells displayed concentration-dependent [Ca2+]i responses when stimulated with ATP (EC50 = 0.86 microM), UTP (EC50 = 1.6 microM) and 2MeSATP (EC50 = 0.38 microM). 2MeSATP behaved as a partial agonist. Ap4A and ADP[beta S] also raised the [Ca2+]i in endothelial cells, whereas AMP and pp[CH2]pA were ineffective. Lowering extracellular free Ca2+ to around 100 nM did not affect the peak ATP-evoked [Ca2+]i rise in these cells. It is concluded that different purinoceptor subtypes are heterogeneously distributed among the major cell types of the adrenal medulla. An intracellular Ca(2+)-releasing P2U-type purinoceptor is specifically localized to adrenal endothelial cells, while a subpopulation of chromaffin cells expresses a non-P2X, non-P2Y subtype exclusively coupled to Ca2+ influx.

MeSH Terms
Adenosine Triphosphate/pharmacology,physiology Adrenal Glands/cytology,metabolism Animals Calcium/metabolism Cattle Cells, Cultured Chromaffin System/metabolism Cytophotometry Endothelium/cytology,metabolism Fura-2 Receptors, Purinergic/drug effects
Chemicals
Receptors, Purinergic Adenosine Triphosphate Calcium Fura-2
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Castro E
Department of Biochemistry, University of Coimbra, Portugal.
Tomé A R
Miras-Portugal M T
Rosário L M
References (55)
55 references, click to expand
  1. An ATP-activated conductance in pheochromocytoma cells and its suppression by extracellular calcium.
    J Physiol. 1990 Sep;428:257-72 PMID: 2231413
  2. Receptor-mediated calcium influx in PC12 cells. ATP and bradykinin activate two independent pathways.
    J Biol Chem. 1990 Nov 25;265(33):20351-5 PMID: 2122973
  3. Multiple actions of extracellular ATP on calcium currents in cultured bovine chromaffin cells.
    Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1261-5 PMID: 1847515
  4. Changes in adrenal oxygen consumption during catecholamine secretion in anesthetized dogs.
    Am J Physiol. 1990 Sep;259(3 Pt 2):H681-8 PMID: 2396681
  5. Evidence for a nucleotide receptor on adrenal medullary endothelial cells linked to phospholipase C and phospholipase D.
    Br J Pharmacol. 1993 Apr;108(4):1031-7 PMID: 8485616
  6. Localization and heterogeneity of agonist-induced changes in cytosolic calcium concentration in single bovine adrenal chromaffin cells from video imaging of fura-2.
    EMBO J. 1989 Feb;8(2):401-11 PMID: 2721487
  7. Comparison of P2 purinergic receptors of aortic endothelial cells with those of adrenal medulla: evidence for heterogeneity of receptor subtype and of inositol phosphate response.
    Mol Pharmacol. 1990 Jul;38(1):84-91 PMID: 2164632
  8. Regulation of bradykinin- and ATP-activated Ca(2+)-permeable channels in rat pheochromocytoma (PC12) cells.
    J Neurosci. 1991 Dec;11(12):3984-90 PMID: 1660537
  9. Agonist-dependent patterns of cytosolic Ca2+ changes in single bovine adrenal chromaffin cells: relationship to catecholamine release.
    Cell Regul. 1990 Aug;1(9):683-91 PMID: 2078572
  10. ATP-activated channels in excitable cells.
    Ion Channels. 1990;2:169-203 PMID: 1715204
  11. Guanine nucleotide-sensitive interaction of a radiolabeled agonist with a phospholipase C-linked P2y-purinergic receptor.
    J Biol Chem. 1989 Apr 15;264(11):6202-6 PMID: 2495280
  12. Adenosine triphosphate stimulates inositol phospholipid metabolism and prostacyclin formation in adrenal medullary endothelial cells by means of P2-purinergic receptors.
    Proc Natl Acad Sci U S A. 1987 Aug;84(16):5630-4 PMID: 3039498
  13. Characteristics of the P2 purinoceptor that mediates prostacyclin production by pig aortic endothelial cells.
    Eur J Pharmacol. 1987 Feb 10;134(2):199-209 PMID: 3552706
  14. High external Ca2+ levels trigger membrane potential oscillations in mouse pancreatic beta-cells during blockade of K(ATP) channels.
    Biochem Biophys Res Commun. 1992 Sep 16;187(2):872-9 PMID: 1530642
  15. ATP-activated single-channel currents recorded from cell-free patches of pheochromocytoma PC12 cells.
    Neurosci Lett. 1990 Oct 30;119(1):5-8 PMID: 1711181
  16. ATP receptor. A putative receptor-operated channel in PC-12 cells.
    J Biol Chem. 1991 Sep 25;266(27):17990-4 PMID: 1655734
  17. A METHOD FOR DIFFERENTIATING BETWEEN NORADRENALINE- AND ADRENALINE-STORING CELLS IN THE LIGHT AND ELECTRON MICROSCOPE.
    Nature. 1964 Mar 21;201:1240-2 PMID: 14151394
  18. Involvement of inositol 1,4,5-trisphosphate and calcium in the action of adenine nucleotides on aortic endothelial cells.
    J Biol Chem. 1987 Dec 25;262(36):17461-6 PMID: 3500950
  19. Bursting electrical activity in pancreatic beta-cells: evidence that the channel underlying the burst is sensitive to Ca2+ influx through L-type Ca2+ channels.
    Pflugers Arch. 1993 Sep;424(5-6):439-47 PMID: 7504808
  20. Vasomotor activity of diadenosine triphosphate and diadenosine tetraphosphate in isolated arteries.
    Am J Physiol. 1988 May;254(5 Pt 2):H828-32 PMID: 3364587
  21. Characterization of two chromaffin cell populations isolated from bovine adrenal medulla.
    J Neurochem. 1991 Aug;57(2):363-9 PMID: 2072090
  22. Stimulation by ATP of inositol trisphosphate accumulation and calcium mobilization in cultured adrenal chromaffin cells.
    J Neurochem. 1989 Feb;52(2):441-7 PMID: 2783453
  23. Cellular responses to Ca2+ from extracellular and intracellular sources are different as shown by simultaneous measurements of cytosolic Ca2+ and secretion from bovine chromaffin cells.
    Proc Natl Acad Sci U S A. 1989 Dec;86(24):9881-5 PMID: 2602380
  24. ATP-activated channels gate calcium entry in single smooth muscle cells dissociated from rabbit ear artery.
    J Physiol. 1989 Dec;419:689-701 PMID: 2559977
  25. The biogenesis of adrenal chromaffin granules.
    Neuroscience. 1977;2(5):657-83 PMID: 593550
  26. Modulation of intracellular pH by secretagogues and the Na+/H+ antiporter in cultured bovine chromaffin cells.
    Neuroscience. 1991;41(1):269-76 PMID: 1647503
  27. Characterization of prostaglandin E2-induced Ca2+ mobilization in single bovine adrenal chromaffin cells by digital image microscopy.
    J Neurochem. 1991 Feb;56(2):531-40 PMID: 1671085
  28. Calcium transients in single adrenal chromaffin cells detected with aequorin.
    FEBS Lett. 1987 Jan 19;211(1):44-8 PMID: 3803586
  29. Characterization of ATP receptor which mediates norepinephrine release in PC12 cells.
    Biochim Biophys Acta. 1992 Sep 9;1136(3):283-9 PMID: 1325838
  30. Extracellular hydrolysis of diadenosine polyphosphates, ApnA, by bovine chromaffin cells in culture.
    Arch Biochem Biophys. 1992 Aug 15;297(1):176-83 PMID: 1322112
  31. Presence of ectonucleotidases in cultured chromaffin cells: hydrolysis of extracellular adenine nucleotides.
    Arch Biochem Biophys. 1990 May 15;279(1):37-44 PMID: 2159757
  32. A comparison of bradykinin, angiotensin II and muscarinic stimulation of cultured bovine adrenal chromaffin cells.
    Biosci Rep. 1989 Apr;9(2):243-52 PMID: 2548638
  33. Extracellular ATP: effects, sources and fate.
    Biochem J. 1986 Jan 15;233(2):309-19 PMID: 3006665
  34. Ca(2+)-stores mobilization by diadenosine tetraphosphate, Ap4A, through a putative P2Y purinoceptor in adrenal chromaffin cells.
    Br J Pharmacol. 1992 Aug;106(4):833-7 PMID: 1393282
  35. Adrenergic receptors of adrenal medullary vasculature.
    Am J Physiol. 1989 Jan;256(1 Pt 2):H233-9 PMID: 2912187
  36. Real-time measurements of acetylcholine-induced release of ATP from bovine medullary chromaffin cells.
    FEBS Lett. 1985 Jun 17;185(2):323-7 PMID: 3996607
  37. Stimulation of formation of inositol phosphates in primary cultures of bovine adrenal chromaffin cells by angiotensin II, histamine, bradykinin, and carbachol.
    J Neurochem. 1988 Aug;51(2):634-41 PMID: 2839623
  38. Endothelial cells from bovine adrenal medulla develop capillary-like growth patterns in culture.
    Proc Natl Acad Sci U S A. 1985 Jul;82(14):4702-6 PMID: 3927288
  39. Regulation of P2y-purinoceptor-mediated prostacyclin release from human endothelial cells by cytoplasmic calcium concentration.
    Br J Pharmacol. 1988 Dec;95(4):1181-90 PMID: 3064854
  40. A new generation of Ca2+ indicators with greatly improved fluorescence properties.
    J Biol Chem. 1985 Mar 25;260(6):3440-50 PMID: 3838314
  41. Pharmacology and electrophysiology of ATP-activated ion channels.
    Trends Pharmacol Sci. 1992 Mar;13(3):87-90 PMID: 1374198
  42. Extracellular ATP and cell signalling.
    Biochim Biophys Acta. 1992 Feb 19;1134(1):31-45 PMID: 1311958
  43. Ca2+ influx induced by the Ca(2+)-ATPase inhibitors 2,5-di-(t-butyl)-1,4-benzohydroquinone and thapsigargin in bovine adrenal chromaffin cells.
    Biochem J. 1992 Dec 1;288 ( Pt 2):457-63 PMID: 1463451
  44. Stimulus-secretion coupling in vascular endothelial cells.
    Annu Rev Physiol. 1990;52:661-74 PMID: 1691907
  45. Voltage-sensitive calcium flux into bovine chromaffin cells occurs through dihydropyridine-sensitive and dihydropyridine- and omega-conotoxin-insensitive pathways.
    Neuroscience. 1989;29(3):735-47 PMID: 2739907
  46. Characterization of P2x-receptors in rabbit isolated ear artery.
    Br J Pharmacol. 1990 Nov;101(3):640-4 PMID: 2076482
  47. Carbachol induced release of diadenosine polyphosphates--Ap4A and Ap5A--from perfused bovine adrenal medulla and isolated chromaffin cells.
    Life Sci. 1991;48(24):2317-24 PMID: 1646361
  48. Preparation of solutions with free calcium concentration in the nanomolar range using 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid.
    Anal Biochem. 1991 Feb 15;193(1):61-71 PMID: 1904204
  49. Further subclassification of ATP receptors based on agonist studies.
    Trends Pharmacol Sci. 1991 Apr;12(4):137-41 PMID: 2063479
  50. A dual function for adenosine 5'-triphosphate in the regulation of vascular tone. Excitatory cotransmitter with noradrenaline from perivascular nerves and locally released inhibitory intravascular agent.
    Circ Res. 1986 Mar;58(3):319-30 PMID: 3013455
  51. P2 purinoceptors on vascular endothelial cells: physiological significance and transduction mechanisms.
    Trends Pharmacol Sci. 1990 Jan;11(1):34-7 PMID: 2408210
  52. Is there a basis for distinguishing two types of P2-purinoceptor?
    Gen Pharmacol. 1985;16(5):433-40 PMID: 2996968
  53. Spontaneous and secretagogue-induced changes in cytosolic free Ca concentration measured by microfluorimetry with fura-2 on single bovine adrenal chromaffin cells.
    Jpn J Physiol. 1989;39(5):687-701 PMID: 2482378
  54. Exogenous ATP raises cytoplasmic free calcium in fura-2 loaded piglet aortic endothelial cells.
    FEBS Lett. 1986 Oct 20;207(1):95-9 PMID: 3533627
  55. The blood supply of the mammalian adrenal medulla: a comparative study.
    J Anat. 1976 Dec;122(Pt 3):539-51 PMID: 795802
Article Info
Journal
Pflugers Archiv : European journal of physiology
Abbr.
Pflugers Arch
ISSN
0031-6768
DOI
10.1007/BF00378530
Published
1994-04-00
Pages
524-33
Language
English
Region
Germany
NLM ID
0154720
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00378530 View journal details
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