Phorbol diester enhances calcium ionophore A23187-induced [3H]acetate incorporation into platelet-activating factor in murine macrophages: predominant incorporation into 1-O-acyl-2-acetyl-sn-glycero-3-phosphocholine

Journal of Cellular Biochemistry
H E Wey

Abstract

Pretreatment of macrophages with 12-O-tetradecanoylphorbol-13-acetate (TPA) has been shown to enhance the release of arachidonic acid from cell phospholipids in response to agonist stimulation. This study describes the ability of TPA to also alter calcium ionophore A23187-induced incorporation of [3H]acetate into platelet activating factor (PAF). Cultured murine peritoneal macrophages were preincubated with [3H]acetate (25 muCi) and TPA (10 ng/ml) for 10 min, and subsequently incubated with 0.1 microM A23187 for 0.5-10 min. Buffer and cells were then extracted and PAF resolved by normal-phase HPLC. Sequential exposure to TPA and A23187 resulted in a greatly enhanced incorporation (11,861 dpm/10(6) cells) of [3H]acetate into PAF compared to TPA alone, which did not significantly influence [3H]acetate incorporation into PAF, and 0.1 microM A23187, which induced minimal incorporation (688 dpm/10(6) cells). Macrophage-produced [3H]PAF was resolved by HPLC, extracted, treated with phospholipase-C, and acetylated to facilitate quantitation of 1-O-alkyl-2-acetyl-GPC (PAF) from 1-O-acyl-2-acetyl-GPC (acylPAF). A23187 alone (1 microM) produced 72% 1-O-acyl-2-[3H]acetyl-GPC, and A23187 (0.1 microM) following TPA pretreatment produced 81%...Continue Reading

References

Jan 1, 1986·Annual Review of Biochemistry·D J Hanahan
Nov 14, 1986·Biochemical and Biophysical Research Communications·H E Wey, C S Baxter
Jul 31, 1985·Biochemical and Biophysical Research Communications·A Mobley, H H Tai
Mar 21, 1986·Biochimica Et Biophysica Acta·M SaitoA Rosenberg
May 16, 1985·Biochemical and Biophysical Research Communications·K SatouchiK Saito
Aug 1, 1959·Canadian Journal of Biochemistry and Physiology·E G BLIGH, W J DYER

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Citations

Jun 19, 1991·Biochimica Et Biophysica Acta·M TriggianiF H Chilton

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