Arachidonic acid metabolism during antigen and ionophore activation of the mouse bone marrow derived mast cell

Tsutomu Nakamura, Alfred N. Fonteh, Walter C. Hubbard, Massimo Triggiani, Naoki Inagaki, Teruko Ishizaka, Floyd H. Chilton

Research output: Contribution to journalArticle

Abstract

This study has examined the metabolism of arachidonic acid in the mouse bone marrow-derived mast cell (BMMC) during immunologic and nonimmunologic activation. The predominant pools of endogenous arachidonate in the mast cells were found in ethanolamine (46%), choline (39%) and inositol (14%) containing glycerolipids. Initial studies established conditions where equilibrium labelling of these major phospholipids in the BMMC could be reached. Upon challenge, arachidonate was lost from all major phospholipid classes (phosphatidylethanolamine > phosphatidylcholine > phosphatidylinositol). There was a small but significant increase in the amount of label associated with phosphatidic acid during cell activation. Arachidonate was distributed among 1-acyl, 1-alkyl and 1-alk-1-enyl-linked subclasses of PC and PE. The rank order of loss of labelled arachidonate from the major PE and PC subclasses during antigen and ionophore activation was 1-alk-enyl-2-arachidonoyl-GPE > 1-acyl-2-arachidonoyl-GPC > 1-acyl-2-arachidonoyl-GPE > 1-alkyl-2-arachidonoyl-GPC. Labelled products released into the supernatant fluids and free arachidonic acid within the cell accounted for the bulk of arachidonate lost from phospholipids. Labelled products in the supernatant fluids were composed of LTB4, LTC4, PGD2 and free arachidonic acid. BMMC phospholipids were also labelled for 24 hr with [3H]choline, [3H]myoinositol or [14H]ethanolamine and labelled 2-lyso phospholipids were measured after cell activation. Radioactivity in lysophospholipids from PC, PE and PI increased significantly between 30 s and 2 min after antigen activation and then declined. Taken together, these studies suggest that arachidonate is mobilized predominantly from PE and in particular 1-alk-1-enyl-2-arachidonoyl-GPE by the direct removal of arachidonate from the sn-2 position of the molecule. Most of this arachidonate is then released from cells as eicosanoids or free fatty acid.

Original languageEnglish (US)
Pages (from-to)191-200
Number of pages10
JournalBiochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism
Volume1085
Issue number2
DOIs
StatePublished - Sep 11 1991

Keywords

  • (Mast cell)
  • Antigen
  • Arachidonic acid metabolism
  • GC-MS
  • Ionophore A23187 Immunological challenge

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Endocrinology

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