Home LiteratureArticle Details
PMID: 7935854 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A1 adenosine receptors expressed in CHO-cells couple to adenylyl cyclase and to phospholipase C.

Naunyn-Schmiedeberg's archives of pharmacology ·Vol. 350 ·No. 1 ·1994-07-00 ·Pages 49-56

Freund S, Ungerer M, Lohse MJ

Abstract

A1 adenosine receptors are in general coupled to inhibition of adenylyl cyclase, but have more recently been reported to be capable of also activating phospholipase C. The present study was done in order to investigate whether these different effects can be elicited by a single A1 receptor, or whether A1 receptor subtypes have to be invoked. The cDNA of a rat brain A1 adenosine receptor was stably expressed in CHO-cells, resulting in clones with varying receptor densities; a clone expressing 1.9 pmol receptors/mg membrane protein was used for further characterization. The ligand binding properties of the expressed receptors were typical for the rat A1 adenosine receptor. A1 receptor agonists caused a concentration-dependent inhibition of adenylyl cyclase activity in the membranes, with maximal inhibition by 70%. A1 receptor stimulation also caused concentration-dependent stimulation of inositol phosphate generation in these cells, with maximal effects of 300%. Both adenylyl cyclase inhibition and enhancement of inositol phosphate generation were essentially abolished after pretreatment of the cells with pertussis toxin. These results indicate that a single A1 adenosine receptor can couple to two effector pathways, and that both effectors are activated via pertussis toxin sensitive G proteins.

MeSH Terms
Adenylate Cyclase Toxin Adenylyl Cyclases/metabolism Animals Binding, Competitive Brain/drug effects,metabolism CHO Cells Cell Membrane/metabolism Cricetinae Cricetulus Inositol Phosphates/metabolism Pertussis Toxin Purinergic P1 Receptor Agonists Purinergic P1 Receptor Antagonists Rats Receptors, Purinergic P1/metabolism Recombinant Proteins/metabolism Signal Transduction Type C Phospholipases/metabolism Virulence Factors, Bordetella/pharmacology Xanthines/metabolism,pharmacology
Chemicals
Adenylate Cyclase Toxin Inositol Phosphates Purinergic P1 Receptor Agonists Purinergic P1 Receptor Antagonists Receptors, Purinergic P1 Recombinant Proteins Virulence Factors, Bordetella Xanthines 1,3-dipropyl-8-cyclopentylxanthine Pertussis Toxin Type C Phospholipases Adenylyl Cyclases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Freund S
Laboratorium für molekulare Biologie der Universität München, Martinsried, Germany.
Ungerer M
Lohse M J
References (54)
54 references, click to expand
  1. Guanine nucleotide and cation regulation of the binding of [3H]cyclohexyladenosine and [3H]diethylphenylxanthine to adenosine A1 receptors in brain membranes.
    Mol Pharmacol. 1982 Mar;21(2):329-35 PMID: 6285169
  2. Structure and function of A1 adenosine receptors.
    FASEB J. 1991 Sep;5(12):2668-76 PMID: 1916091
  3. Mechanism of A2 adenosine receptor activation. I. Blockade of A2 adenosine receptors by photoaffinity labeling.
    Mol Pharmacol. 1991 Apr;39(4):517-23 PMID: 2017151
  4. Agonist photoaffinity labeling of A1 adenosine receptors: persistent activation reveals spare receptors.
    Mol Pharmacol. 1986 Oct;30(4):403-9 PMID: 3020390
  5. Calcium-dependent currents in cultured rat dorsal root ganglion neurones are inhibited by an adenosine analogue.
    J Physiol. 1986 Apr;373:47-61 PMID: 2427698
  6. Multiple second messenger pathways of alpha-adrenergic receptor subtypes expressed in eukaryotic cells.
    J Biol Chem. 1990 Jan 5;265(1):63-9 PMID: 2152928
  7. Regulation of purified subtypes of phosphatidylinositol-specific phospholipase C beta by G protein alpha and beta gamma subunits.
    J Biol Chem. 1993 May 5;268(13):9667-74 PMID: 8387502
  8. Stimulation of adenosine A1 receptors and bradykinin receptors, which act via different G proteins, synergistically raises inositol 1,4,5-trisphosphate and intracellular free calcium in DDT1 MF-2 smooth muscle cells.
    Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7330-4 PMID: 1323831
  9. Transfected m2 muscarinic acetylcholine receptors couple to G alpha i2 and G alpha i3 in Chinese hamster ovary cells. Activation and desensitization of the phospholipase C signaling pathway.
    J Biol Chem. 1993 Mar 15;268(8):5676-85 PMID: 8449930
  10. Interactions between intracellular cyclic AMP and agonist-induced inositol phospholipid breakdown in isolated gastric mucosal cells of the rat.
    Naunyn Schmiedebergs Arch Pharmacol. 1987 Nov;336(5):471-7 PMID: 2830543
  11. Effects of adenosine on histamine release from human lung fragments.
    Int Arch Allergy Immunol. 1992;98(1):50-6 PMID: 1378041
  12. Adenosine regulates the Ca2+ sensitivity of mast cell mediator release.
    Proc Natl Acad Sci U S A. 1988 Dec;85(23):8875-9 PMID: 2461558
  13. Regulation of GH3-cell function via adenosine A1 receptors. Inhibition of prolactin release, cyclic AMP production and inositol phosphate generation.
    Biochem J. 1988 Oct 1;255(1):69-77 PMID: 2848512
  14. An M2 muscarinic receptor subtype coupled to both adenylyl cyclase and phosphoinositide turnover.
    Science. 1987 Oct 30;238(4827):672-5 PMID: 2823384
  15. Adenosine A1 receptor gene structure and regulation in normotensive and spontaneously hypertensive rats.
    Eur J Pharmacol. 1992 Aug 3;226(4):381-2 PMID: 1397063
  16. Subclasses of external adenosine receptors.
    Proc Natl Acad Sci U S A. 1980 May;77(5):2551-4 PMID: 6248853
  17. Binding of the A1-selective adenosine antagonist 8-cyclopentyl-1,3-dipropylxanthine to rat brain membranes.
    Naunyn Schmiedebergs Arch Pharmacol. 1987 Jan;335(1):59-63 PMID: 3574492
  18. Is the adenosine receptor modulation of histamine-induced accumulation of inositol phosphates in cerebral cortical slices mediated by effects on calcium ion fluxes?
    J Neurochem. 1990 Oct;55(4):1138-41 PMID: 2118942
  19. ATP and A1 adenosine receptor agonists mobilize intracellular calcium and activate K+ and Cl- currents in normal and cystic fibrosis airway epithelial cells.
    J Biol Chem. 1993 Nov 25;268(33):24779-84 PMID: 8227038
  20. Differential regulation of PI hydrolysis and adenylyl cyclase by muscarinic receptor subtypes.
    Nature. 1988 Aug 4;334(6181):434-7 PMID: 2841607
  21. Two affinity states of Ri adenosine receptors in brain membranes. Analysis of guanine nucleotide and temperature effects on radioligand binding.
    Mol Pharmacol. 1984 Jul;26(1):1-9 PMID: 6087114
  22. Reduction of adenylate cyclase activity in lysates of human platelets by the alpha-adrenergic component of epinephrine.
    J Cyclic Nucleotide Res. 1976 Nov-Dec;2(6):381-92 PMID: 14175
  23. Activation of phospholipase C via adenosine receptors provides synergistic signals for secretion in antigen-stimulated RBL-2H3 cells. Evidence for a novel adenosine receptor.
    J Biol Chem. 1990 Jan 15;265(2):745-53 PMID: 2295618
  24. Isolated atrial myocytes: adenosine and acetylcholine increase potassium conductance.
    Am J Physiol. 1983 May;244(5):H734-7 PMID: 6846564
  25. Guanine nucleotides and adenosine "R(i)'-site analogues stimulate the membrane-bound low-K(m) cyclic AMP phosphodiesterase of rat adipocytes.
    FEBS Lett. 1984 Aug 6;173(2):385-8 PMID: 6086407
  26. Stable overexpression of human beta 2-adrenergic receptors in mammalian cells.
    Naunyn Schmiedebergs Arch Pharmacol. 1992 Apr;345(4):444-51 PMID: 1320210
  27. Adenosine regulates via two different types of receptors, the accumulation of cyclic AMP in cultured brain cells.
    J Neurochem. 1979 Nov;33(5):999-1005 PMID: 228008
  28. Differential coupling of dopaminergic D2 receptors expressed in different cell types. Stimulation of phosphatidylinositol 4,5-bisphosphate hydrolysis in LtK- fibroblasts, hyperpolarization, and cytosolic-free Ca2+ concentration decrease in GH4C1 cells.
    J Biol Chem. 1990 Jun 25;265(18):10320-6 PMID: 2162345
  29. Adenosine A1-receptor stimulation of inositol phospholipid hydrolysis and calcium mobilisation in DDT1 MF-2 cells.
    Br J Pharmacol. 1992 May;106(1):215-21 PMID: 1324066
  30. A1 adenosine receptors inhibit chloride transport in the shark rectal gland. Dissociation of inhibition and cyclic AMP.
    J Clin Invest. 1990 May;85(5):1629-36 PMID: 1970583
  31. The orphan receptor cDNA RDC7 encodes an A1 adenosine receptor.
    EMBO J. 1991 Jul;10(7):1677-82 PMID: 1646713
  32. Isozyme-selective stimulation of phospholipase C-beta 2 by G protein beta gamma-subunits.
    Nature. 1992 Dec 17;360(6405):684-6 PMID: 1465133
  33. Validation and statistical analysis of a computer modeling method for quantitative analysis of radioligand binding data for mixtures of pharmacological receptor subtypes.
    Mol Pharmacol. 1982 Jan;21(1):5-16 PMID: 6982395
  34. Effector coupling mechanisms of the cloned 5-HT1A receptor.
    J Biol Chem. 1989 Sep 5;264(25):14848-52 PMID: 2549039
  35. Molecular cloning and characterization of a rat A1-adenosine receptor that is widely expressed in brain and spinal cord.
    Mol Endocrinol. 1991 Aug;5(8):1037-48 PMID: 1658635
  36. Effects of adenosine on cAMP production and cytosolic Ca2+ in cultured rabbit medullary thick limb cells.
    Am J Physiol. 1991 Jan;260(1 Pt 1):C143-50 PMID: 1846267
  37. Role of guanine nucleotide-binding protein in the regulation by adenosine of cardiac potassium conductance and force of contraction. Evaluation with pertussis toxin.
    Naunyn Schmiedebergs Arch Pharmacol. 1986 Apr;332(4):403-5 PMID: 3090454
  38. ATP and its metabolite adenosine act synergistically to mobilize intracellular calcium via the formation of inositol 1,4,5-trisphosphate in a smooth muscle cell line.
    J Biol Chem. 1992 Aug 15;267(23):16081-7 PMID: 1322890
  39. Subunits beta gamma of heterotrimeric G protein activate beta 2 isoform of phospholipase C.
    Nature. 1992 Dec 17;360(6405):686-9 PMID: 1465134
  40. Receptor-to-effector signaling through G proteins: roles for beta gamma dimers as well as alpha subunits.
    Cell. 1992 Dec 24;71(7):1069-72 PMID: 1335363
  41. Stimulation of adenosine receptor enhances alpha 1-adrenergic receptor-mediated activation of phospholipase C and Ca2+ mobilization in a pertussis toxin-sensitive manner in FRTL-5 thyroid cells.
    FEBS Lett. 1989 May 8;248(1-2):145-9 PMID: 2542083
  42. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  43. Photoaffinity labeling of A1-adenosine receptors.
    J Biol Chem. 1985 Nov 25;260(27):14659-64 PMID: 2997218
  44. Activation of phospholipase C isozymes by G protein beta gamma subunits.
    J Biol Chem. 1993 Mar 5;268(7):4573-6 PMID: 8383116
  45. Adenosine stimulates guanylate cyclase activity in vascular smooth muscle cells.
    J Biol Chem. 1987 May 5;262(13):6296-300 PMID: 2883183
  46. 8-Cyclopentyl-1,3-dipropylxanthine (DPCPX)--a selective high affinity antagonist radioligand for A1 adenosine receptors.
    Naunyn Schmiedebergs Arch Pharmacol. 1987 Aug;336(2):204-10 PMID: 2825043
  47. Effects of carbachol and (-)-N6-phenylisopropyladenosine on myocardial inositol phosphate content and force of contraction.
    Br J Pharmacol. 1990 Dec;101(4):829-34 PMID: 2085707
  48. Accumulations of inositol phosphates and cyclic AMP in brain slices: synergistic interactions of histamine and 2-chloroadenosine.
    Eur J Pharmacol. 1986 Mar 11;122(1):45-50 PMID: 3007178
  49. Dual actions of adenosine on rat peritoneal mast cells.
    Naunyn Schmiedebergs Arch Pharmacol. 1987 May;335(5):555-60 PMID: 2441269
  50. Cloning and expression of an A1 adenosine receptor from rat brain.
    Mol Pharmacol. 1991 Jul;40(1):1-7 PMID: 1857334
  51. Functional expression of human 5-HT1A receptors and differential coupling to second messengers in CHO cells.
    Naunyn Schmiedebergs Arch Pharmacol. 1992 Aug;346(2):127-37 PMID: 1448178
  52. Adenosine-activated potassium conductance in cultured striatal neurons.
    Proc Natl Acad Sci U S A. 1985 Jul;82(14):4857-61 PMID: 2991897
  53. Interactions of the bovine brain A1-adenosine receptor with recombinant G protein alpha-subunits. Selectivity for rGi alpha-3.
    J Biol Chem. 1991 Sep 25;266(27):17778-83 PMID: 1917922
  54. Cyclic AMP differentiates two separate but interacting pathways of phosphoinositide hydrolysis in the DDT1-MF2 smooth muscle cell line.
    Mol Pharmacol. 1992 Mar;41(3):587-97 PMID: 1312218
Article Info
Journal
Naunyn-Schmiedeberg's archives of pharmacology
Abbr.
Naunyn Schmiedebergs Arch Pharmacol
ISSN
0028-1298
DOI
10.1007/BF00180010
Published
1994-07-00
Pages
49-56
Language
English
Region
Germany
NLM ID
0326264
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00180010 View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2