Home LiteratureArticle Details
PMID: 1850985 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Determination of mass changes in phosphatidylinositol 4,5-bisphosphate and evidence for agonist-stimulated metabolism of inositol 1,4,5-trisphosphate in airway smooth muscle.

The Biochemical journal ·Vol. 275 ( Pt 2) ·1991-04-15 ·Pages 373-9

Chilvers ER, Batty IH, Challiss RA, Barnes PJ, Nahorski SR

Abstract

Stimulation of muscarinic receptors in bovine tracheal smooth muscle (BTSM) causes a sustained increase in muscle tone, but a transient increase in the second messenger Ins(1,4,5)P3. To examine whether this brief increase in Ins(1,4,5)P3 mass results from transient formation or is due to agonist-stimulation of Ins(1,4,5)P3 metabolism, we have studied the relationship between mass changes in PtdIns(4,5)P2 and Ins(1,4,5)P3 accumulation, and changes in [3H]InsP3, [3H]PtdIns, [3H]PtdInsP1 and [3H]PtdInsP2 in carbachol-stimulated myo-[3H]inositol-prelabelled BTSM slices. Carbachol (0.1 mM) caused a rapid transient increase in Ins(1,4,5)P3 concentration (basal, 12.9 +/- 0.8 pmol/mg of protein; 5 s carbachol treatment, 27.1 +/- 1.5 pmol/mg of protein), with values returning to basal levels by 30 s, but a sustained accumulation of total [3H]InsP3s, with [3H]Ins(1,3,4)P3 being the predominant isomer present at later time points. In contrast, PtdIns(4,5)P2 mass, determined by radioreceptor assay of Ins(1,4,5)P3 in desalted alkaline hydrolysates of acidified chloroform/methanol tissue extracts, declined rapidly (basal, 941 +/- 22 pmol/mg of protein; 120 s carbachol, 365 +/- 22 pmol/mg of protein; t1/2 14 s) and remained at this new steady-state level for at least 20 min in the continued presence of carbachol. Addition of 10 microM-atropine 2 min after carbachol caused a prompt return of PtdIns(4,5)P2 concentration to prestimulated values (t1/2 210 s). Ongoing resynthesis of PtdIns(4,5)P2 after carbachol stimulation was demonstrated in [3H]inositol-labelled tissue by observing a persistent increase in the specific radioactivity of [3H]PtdInsP2, shown to be exclusively [3H]PtdIns(4,5)P2, over a 10 min period. These findings strongly suggest the occurrence of persistent receptor-mediated increases in PtdIns(4,5)P2 hydrolysis and Ins(1,4,5)P3 formation which, in conjunction with the transient accumulation of Ins(1,4,5)P3 observed, provide evidence that regulation of the metabolism of Ins(1,4,5)P3 is a major determinant of Ins(1,4,5)P3 concentration in this tissue under agonist-stimulated conditions.

MeSH Terms
Animals Carbachol/pharmacology Cattle Chromatography, High Pressure Liquid In Vitro Techniques Inositol 1,4,5-Trisphosphate/metabolism Inositol Phosphates/isolation & purification,metabolism Kinetics Muscle, Smooth/drug effects,metabolism Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols/metabolism Radioisotope Dilution Technique Trachea/drug effects,metabolism Tritium
Chemicals
Inositol Phosphates Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols Tritium Inositol 1,4,5-Trisphosphate Carbachol
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Chilvers E R
Department of Pharmacology and Therapeutics, University of Leicester, U.K.
Batty I H
Challiss R A
Barnes P J
Nahorski S R
References (61)
61 references, click to expand
  1. Characterization of agonist-stimulated incorporation of myo-[3H]inositol into inositol phospholipids and [3H]inositol phosphate formation in tracheal smooth muscle.
    Biochem J. 1989 Sep 15;262(3):739-46 PMID: 2556108
  2. Mass changes of inositol(1,4,5)trisphosphate in trachealis muscle following agonist stimulation.
    Eur J Pharmacol. 1989 May 30;164(3):587-90 PMID: 2548874
  3. Inositol phosphates and cell signalling.
    Nature. 1989 Sep 21;341(6239):197-205 PMID: 2550825
  4. The identification of a novel inositol lipid, phosphatidylinositol trisphosphate (PIP3), in rat cerebrum using in vivo techniques.
    Biochem Biophys Res Commun. 1989 Sep 15;163(2):995-1001 PMID: 2551281
  5. Transient increase in phosphatidylinositol 3,4-bisphosphate and phosphatidylinositol trisphosphate during activation of human neutrophils.
    J Biol Chem. 1989 Sep 15;264(26):15668-73 PMID: 2549071
  6. Quantitative changes in polyphosphoinositides 1,2-diacylglycerol and inositol 1,4,5-trisphosphate by platelet-derived growth factor and prostaglandin F2 alpha.
    J Biol Chem. 1989 Sep 5;264(25):14985-9 PMID: 2549046
  7. Metabolism of the inositol phosphates produced upon receptor activation.
    Biochem J. 1989 Jun 1;260(2):313-24 PMID: 2548474
  8. Neurotransmitter receptors and phosphoinositide turnover.
    Annu Rev Pharmacol Toxicol. 1989;29:71-110 PMID: 2543275
  9. Metabolic and structural evidence for the existence of a third species of polyphosphoinositide in cells: D-phosphatidyl-myo-inositol 3-phosphate.
    Biochem J. 1989 Apr 1;259(1):267-76 PMID: 2541684
  10. Chemoattractant receptor promotion of Ca2+ influx across the plasma membrane of HL-60 cells. A role for cytosolic free calcium elevations and inositol 1,3,4,5-tetrakisphosphate production.
    J Biol Chem. 1989 May 5;264(13):7251-61 PMID: 2540183
  11. Development of a novel, Ins(1,4,5)P3-specific binding assay. Its use to determine the intracellular concentration of Ins(1,4,5)P3 in unstimulated and vasopressin-stimulated rat hepatocytes.
    Cell Signal. 1989;1(2):147-56 PMID: 2642027
  12. Formation of [3H]inositol metabolites in rat hippocampal formation slices prelabelled with [3H]inositol and stimulated with carbachol.
    J Neurochem. 1989 Mar;52(3):713-21 PMID: 2783959
  13. A simple enzymic method to separate [3H]inositol 1,4,5- and 1,3,4-trisphosphate isomers in tissue extracts.
    Biochem J. 1989 May 15;260(1):283-6 PMID: 2789038
  14. Accumulation of inositol polyphosphate isomers in agonist-stimulated cerebral-cortex slices. Comparison with metabolic profiles in cell-free preparations.
    Biochem J. 1989 Feb 15;258(1):23-32 PMID: 2930510
  15. Inositol lipid turnover and compartmentation in canine trachealis smooth muscle.
    Am J Physiol. 1989 Feb;256(2 Pt 1):C375-83 PMID: 2919664
  16. The Croonian lecture, 1988. Inositol lipids and calcium signalling.
    Proc R Soc Lond B Biol Sci. 1988 Sep 22;234(1277):359-78 PMID: 2906130
  17. Mass measurements of inositol(1,4,5)trisphosphate in rat cerebral cortex slices using a radioreceptor assay: effects of neurotransmitters and depolarization.
    Biochem Biophys Res Commun. 1988 Dec 15;157(2):684-91 PMID: 2904813
  18. Inositol phosphates: synthesis and degradation.
    J Biol Chem. 1988 Mar 5;263(7):3051-4 PMID: 3277965
  19. An inositol tetrakisphosphate-containing phospholipid in activated neutrophils.
    Nature. 1988 Jul 28;334(6180):353-6 PMID: 3393226
  20. Inositol 1,2-cyclic 4,5-trisphosphate concentration relative to inositol 1,4,5-trisphosphate in pancreatic minilobules on stimulation with carbamylcholine in the absence of lithium. Possible role as a second messenger in long- but not short-term responses.
    J Biol Chem. 1987 Oct 15;262(29):13892-5 PMID: 3498721
  21. Rapid formation of inositol 1,3,4,5-tetrakisphosphate following muscarinic receptor stimulation of rat cerebral cortical slices.
    Biochem J. 1985 Nov 15;232(1):211-5 PMID: 4084229
  22. Inositol phospholipid levels of rat forebrain obtained by freeze-blowing method.
    Biochim Biophys Acta. 1985 Jul 9;835(2):415-8 PMID: 4005289
  23. Characterisation of stereospecific binding sites for inositol 1,4,5-trisphosphate in airway smooth muscle.
    Br J Pharmacol. 1990 Feb;99(2):297-302 PMID: 2158373
  24. Phosphorylation by protein kinase C inactivates an inositol 1,4,5-trisphosphate 3-kinase purified from human platelets.
    Biochem Biophys Res Commun. 1990 Aug 16;170(3):1371-6 PMID: 2167676
  25. Regulation of D-myo-inositol 1,4,5-trisphosphate 3-kinase by cAMP-dependent protein kinase and protein kinase C.
    J Biol Chem. 1990 Jun 25;265(18):10367-72 PMID: 2162347
  26. Characterization of the muscarinic receptor subtype involved in phosphoinositide metabolism in bovine tracheal smooth muscle.
    Br J Pharmacol. 1990 Feb;99(2):293-6 PMID: 2158372
  27. Heterogeneity of [3H]inositol 1,4,5-trisphosphate binding sites in adrenal-cortical membranes. Characterization and validation of a radioreceptor assay.
    Biochem J. 1990 Jan 15;265(2):421-7 PMID: 2154189
  28. Formation of inositol polyphosphates in airway smooth muscle after muscarinic receptor stimulation.
    J Pharmacol Exp Ther. 1990 Feb;252(2):786-91 PMID: 2313600
  29. Breakdown of polyphosphoinositides and not phosphatidylinositol accounts for muscarinic agonist-stimulated inositol phospholipid metabolism in rat parotid glands.
    Biochem J. 1983 Dec 15;216(3):633-40 PMID: 6320795
  30. Phosphoinositide synthesis and Ca2+ gating in blowfly salivary glands exposed to 5-hydroxytryptamine.
    Biochem J. 1984 Sep 1;222(2):327-34 PMID: 6089767
  31. The inositol trisphosphate phosphomonoesterase of the human erythrocyte membrane.
    Biochem J. 1982 Apr 1;203(1):169-77 PMID: 6285891
  32. Alkaline O leads to N-transacylation. A new method for the quantitative deacylation of phospholipids.
    Biochem J. 1981 Apr 1;195(1):301-6 PMID: 7306057
  33. Calcium-activated hydrolysis of phosphatidyl-myo-inositol 4-phosphate and phosphatidyl-myo-inositol 4,5-bisphosphate in guinea-pig synaptosomes.
    Biochem J. 1978 Nov 15;176(2):541-52 PMID: 217364
  34. Myo-inositol lipids.
    Vitam Horm. 1975;33:529-73 PMID: 180679
  35. Two different G-proteins mediate neuropeptide Y and bradykinin-stimulated phospholipid breakdown in cultured rat sensory neurons.
    J Biol Chem. 1989 May 5;264(13):7317-27 PMID: 2540185
  36. Polyphosphoinositides produced by phosphatidylinositol 3-kinase are poor substrates for phospholipases C from rat liver and bovine brain.
    J Biol Chem. 1989 Oct 25;264(30):17809-15 PMID: 2553693
  37. Electrophysiological and other aspects of excitation-contraction coupling and uncoupling in mammalian airway smooth muscle.
    Life Sci. 1987 Jul 13;41(2):111-32 PMID: 2439865
  38. Inhibition of protein kinase C by staurosporine promotes elevated accumulations of inositol trisphosphates and tetrakisphosphate in human platelets exposed to thrombin.
    J Biol Chem. 1989 Apr 15;264(11):6070-4 PMID: 2703480
  39. 1,2-Diacylglycerol and phorbol ester inhibit agonist-induced formation of inositol phosphates in human platelets: possible implications for negative feedback regulation of inositol phospholipid hydrolysis.
    Proc Natl Acad Sci U S A. 1985 May;82(9):2623-6 PMID: 2986126
  40. Metabolism of inositol 1,4,5-trisphosphate and inositol 1,3,4-trisphosphate in rat parotid glands.
    Biochem J. 1985 Jul 15;229(2):505-11 PMID: 2994638
  41. Is inositol bisphosphate the product of A23187 and carbachol-mediated polyphosphoinositide breakdown in synaptosomes?
    J Neurochem. 1988 Aug;51(2):514-21 PMID: 2839621
  42. Characterization of inositol trisphosphate receptor binding in brain. Regulation by pH and calcium.
    J Biol Chem. 1987 Sep 5;262(25):12132-6 PMID: 3040730
  43. Cyclic AMP-dependent phosphorylation of a brain inositol trisphosphate receptor decreases its release of calcium.
    Proc Natl Acad Sci U S A. 1988 Nov;85(22):8747-50 PMID: 2847175
  44. Protein kinase C phosphorylates human platelet inositol trisphosphate 5'-phosphomonoesterase, increasing the phosphatase activity.
    Cell. 1986 Sep 12;46(6):951-8 PMID: 3019558
  45. Ca2+ regulates the inositol tris/tetrakisphosphate pathway in intact and broken preparations of insulin-secreting RINm5F cells.
    J Biol Chem. 1986 Sep 15;261(26):11931-4 PMID: 3017952
  46. Phosphatidylinositol response to cholinergic agonists in airway smooth muscle: relationship to contraction and muscarinic receptor occupancy.
    J Pharmacol Exp Ther. 1986 Jul;238(1):273-9 PMID: 3014119
  47. The inositol tris/tetrakisphosphate pathway--demonstration of Ins(1,4,5)P3 3-kinase activity in animal tissues.
    Nature. 1986 Apr 17-23;320(6063):631-4 PMID: 3010126
  48. Angiotensin-stimulated production of inositol trisphosphate isomers and rapid metabolism through inositol 4-monophosphate in adrenal glomerulosa cells.
    Proc Natl Acad Sci U S A. 1986 Dec;83(24):9323-7 PMID: 3025836
  49. The metabolism of phosphoinositide-derived messenger molecules.
    Science. 1986 Dec 19;234(4783):1519-26 PMID: 3024320
  50. Carbachol induces a rapid and sustained hydrolysis of polyphosphoinositide in bovine tracheal smooth muscle measurements of the mass of polyphosphoinositides, 1,2-diacylglycerol, and phosphatidic acid.
    J Biol Chem. 1986 Nov 5;261(31):14670-5 PMID: 3021749
  51. Dependence on Ca2+ of the activities of phosphatidylinositol 4,5-bisphosphate phosphodiesterase and inositol 1,4,5-trisphosphate phosphatase in smooth muscles of the porcine coronary artery.
    Biochem J. 1985 Nov 1;231(3):497-503 PMID: 3000351
  52. Requirement of Ca2+ for the production and degradation of inositol 1,4,5-trisphosphate in macrophages.
    Biochim Biophys Acta. 1986 Jan 23;885(1):121-8 PMID: 3002487
  53. Formation and metabolism of [3H]inositol phosphates in AR42J pancreatoma cells. Substance P-induced Ca2+ mobilization in the apparent absence of inositol 1,4,5-trisphosphate 3-kinase activity.
    J Biol Chem. 1988 Oct 25;263(30):15297-303 PMID: 2459122
  54. Inositol 1,3,4,5-tetrakisphosphate and not phosphatidylinositol 3,4-bisphosphate is the probable precursor of inositol 1,3,4-trisphosphate in agonist-stimulated parotid gland.
    Biochem J. 1986 Sep 1;238(2):501-6 PMID: 2432882
  55. Activation of tracheal smooth muscle contraction: synergism between Ca2+ and activators of protein kinase C.
    Proc Natl Acad Sci U S A. 1985 Dec;82(24):8835-9 PMID: 2417230
  56. Formation and metabolism of inositol 1,3,4,5-tetrakisphosphate in liver.
    J Biol Chem. 1986 Jun 25;261(18):8100-3 PMID: 3487541
  57. Isomers of inositol trisphosphate in exocrine pancreas.
    Biochem J. 1986 Sep 15;238(3):825-9 PMID: 3492200
  58. Phorbol 12,13-dibutyrate and 1-oleyl-2-acetyldiacylglycerol stimulate inositol trisphosphate dephosphorylation in human platelets.
    J Biol Chem. 1986 Aug 15;261(23):10493-5 PMID: 3488315
  59. Carbachol-induced protein phosphorylation changes in bovine tracheal smooth muscle.
    J Biol Chem. 1986 Nov 25;261(33):15734-9 PMID: 3782086
  60. A role for inositol 1,4,5-trisphosphate in the initiation of agonist-induced contractions of dog tracheal smooth muscle.
    Br J Pharmacol. 1985 Sep;86(1):191-9 PMID: 3876861
  61. Mass changes in myoinositol trisphosphate in human platelets stimulated by thrombin. Inhibitory effects of phorbol ester.
    J Biol Chem. 1985 Jul 25;260(15):8657-60 PMID: 3874867
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2750373
Published
1991-04-15
Pages
373-9
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1150063
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1042/bj2750373 PMC Full Text 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