Transfer of arachidonic acid to human platelet plasmalogen in response to thrombin

Rittenhouse-Simmons, S; Russell, Francis A.; Deykin, Daniel

doi:10.1016/0006-291x(76)91141-4

Cited by 81 publications

(22 citation statements)

References 6 publications

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“…For studies on the metabolismn of radiolabeled arachi(loniic acid or glycerol, PRP was incuibated with [5,6,8,9,11,12,14,15-3H]arachidonic acid (12.5 ,uCi/30 ml PRP, 72 Ci/mmol) which had been hound to albuumin as described previously (14) (8,19). Radioactivity contained in the lipid spots wvas counted by scintillationi spectrophotometry in H20-AqIuasol (New England Nuclear) (8), and lipid phosphorus was assayed after dligestioni of parallel samiiples (19).…”

Section: Methodsmentioning

confidence: 99%

Production of diglyceride from phosphatidylinositol in activated human platelets.

1979

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Section: Methodsmentioning

confidence: 99%

Production of diglyceride from phosphatidylinositol in activated human platelets.

1979

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“…Blood was drawn from normal donors and the plateletrich plasma incubated with [5,6,8,9,11,12,14,15-3H larachidonic acid (10 nM) or [14C]hydroxytryptamine (creatinine sulfate salt, 0.4 ,uM) as previously described (4). Plateletrich plasma was then filtered through a 0.5-ml layer of 2 mM ethylene glycol-bis(,f-aminoethyl ether) N,N,N'N'-tetraacetate (EGTA)1 on Sepharose 2B equilibrated with a calcium ion-free buffer (pH 7) described elsewhere (6).…”

Section: Methodsmentioning

confidence: 99%

“…Supernatant lipids were also extracted into chloroform/methanol. Lipid chromatography and distribution of radiolabel among the lipid components were determined as previously described (4,5).…”

Section: Methodsmentioning

confidence: 99%

“…In this report we examine whether similar starvation leads to the impairment of thrombin-promoted mobilization of arachidonic acid (4,5) to determine what link may exist between the release reaction and the activation of platelet phospholipase A2. We have utilized the calcium ionophore Received for publication 7 March 1977 and in revised form 11 April 1977. A23187 to determine further whether platelet phospholipase activation is comparably energy-dependent or is a direct consequence of calcium flux.…”

Section: Introductionmentioning

confidence: 99%

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The mobilization of arachidonic acid in platelets exposed to thrombin or ionophore A23187. Effects of adenosine triphosphate deprivation.

Rittenhouse-Simmons¹,

Deykin²

1977

J. Clin. Invest.

116

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A B S T R A C T In studies conducted with human gelfiltered platelets, we have found: (a) that the release of serotonin and transfer of [3H]arachidonic acid from phosphatidylcholine and phosphatidylinositol to plasmalogen phosphatidylethanolamine which are associated with the activation of platelets by thrombin are both strongly dependent upon the presence of metabolic ATP; (b) that serotonin release and arachidonic acid mobilization in labeled phosphatides are promoted by the calcium ionophore A-23187 in media free of calcium ions; (c) that inhibitors of ATP synthesis, while leading to impairment of the release reaction induced by ionophore, do not inhibit ionophore-stimulated mobilization of arachidonic acid.We conclude that the activation of phospholipase A2 responsible for freeing arachidonic acid from platelet phosphatides is solely dependent upon the increased cytoplasmic levels of calcium ions promoted by either ionophore or, in an energy-dependent fashion by thrombin. Phospholipase activation is not a function of latent hydrolytic activity made available by the release reaction.

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“…It was recently suggested that arachidonic acid may play an important role in the activation of lymphocyte [I51 and also arachidonic acid was transfered to plasmalogen from other phospholipid in human platelets incubated with thrombin [16]. In such a Ehrlich ascites tumor cells which proliferate rapidly, arachidonic acid might have a very important role for cell development.…”

Section: Resultsmentioning

confidence: 99%

Turnover Rates of the Molecular Species of Alkenyl Ether Phospholipids of Ehrlich Ascites Tumor Cells

Waku

Nakazawa

1979

Eur J Biochem

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[l -14C]Glycerol was injected into the peritoneal cavity of mice bearing Ehrlich ascites tumor cells and the lipids were extracted from the cells after selected periods. l-O-Alkenyl-2-acyl-3-acetylglycerols were prepared from the ethanolamine phosphoglycerides and fractionated by A,N03-impregnated thin-layer chromatography into five molecular species. Bands 1 -5 obtained in this way can be designated as saturated, monoene, diene, tetraene and hexaene species, respectively. Band 5, composed mainly of hexaene species was predominant among those derived from alkenyl ether phospholipids.The specific radioactivities of the fractionated species were determined. Hexaene molecular species turned over most rapidly and disaturated and tetraene species turned over at a high rate, while monoene and diene molecular species turned over very slowly.The turnover rate of alkenyl ether phospholipids (plasmalogen) determined by the incorporation experiments of 32P,, [l -14C]glycerol and [1-l4C]acetate into the lipids of Ehrlich ascites tumor cells was slower than those of diacyl and alkyl ether phospholipids [l]. However, it has recently been suggested that the plasmalogen may be the donor of the precursors of prostaglandins through the action of plasmalogenase and the subsequent hydrolysis by the lysophospholipase [2,3]. It this is the case, it may be an interesting problem to study the turnover rates of every molecular species of plasmalogen.In a previous paper [4], we determined the turnover rates of the molecular species of alkyl and diacyl glycerophospholipids of Ehrlich ascites tumor cells and it was demonstrated that in these ethanolamine phospholipids, hexaene molecular species showed the highest turnover rate.In this work, [l-14C]glycerol was injected into the peritoneal cavity of mice bearing Ehrlich ascites tumor cells and the incorporation rates of the radioactivity into the molecular species of alkenyl ether phospholipids were estimated and compared with those of alkyl acyl ethanolamine phospholipids. The results of a detailed analysis of the separated species of 1-0-alkenyl phospholipids are also described. phosphoglycerides were fractionated and 1 -radyl-2-acyl-3-acetylglycerol was prepared as described earlier [5]. l-O-Alkenyl-2-acyl-3-acetylglycerol was isolating according to the method of Renkonen [6]. The quantities of the alkenyl and fatty acyl contents were estimated by the spectrophotometric method [7] and the gas-liquid chromatography using 15 : 0 methyl ester as an internal standard, respectively. The molar ratio of alkenyl and ester content was confirmed to be equivalent. The compound was further fractionated by 20 % AgN03-impregnated thin-layer chromatography (Silica gel H) developed with 0.8 % methanol in chloroform. The detection and extraction of individual spot were performed as described previously [4]. A part of the lipid fraction was used for the measurement of the radioactivity and the remaining sample was used for the analysis of fatty chains of positions 1 and 2. The recovery of lipids and radio...

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Transfer of arachidonic acid to human platelet plasmalogen in response to thrombin

Cited by 81 publications

References 6 publications

Production of diglyceride from phosphatidylinositol in activated human platelets.

Production of diglyceride from phosphatidylinositol in activated human platelets.

The mobilization of arachidonic acid in platelets exposed to thrombin or ionophore A23187. Effects of adenosine triphosphate deprivation.

Turnover Rates of the Molecular Species of Alkenyl Ether Phospholipids of Ehrlich Ascites Tumor Cells

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