Lack of Phospholipase D Activity in Chromaffin Cells: Bradykinin‐Stimulated Phosphatidic Acid Formation Involves Phospholipase C in Chromaffin Cells but Phospholipase D in PC12 Cells

John Purkiss, Richard A. Murrin, P. Jane Owen, Michael R. Boarder

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The role of lipid‐bound second messengers in the regulation of neurotransmitter secretion is an important but poorly understood subject. Both bovine adrenal Chromaffin cells and rat phoeochromocytoma (PC12) cells, two widely studied models of neuronal function, respond to bradykinin by generating phosphatidic acid (PA). This putative second messenger may be produced by two receptor‐linked pathways: sequential action of phospholipase C (PLC) and diacylglycerol kinase(DAG kinase), or directly by phospholipase D (PLD). Here we show that bradykinin stimulation of Chromaffin cells prelabelled (24 h) with 32P1 leads to production of [32P]PA which is not affected by 50 mM butanol. However, bradykinin stimulation of PC12 cells leads to [32P]PA formation, all of which is converted to phosphatidylbutanol in the presence of butanol. When Chromaffin cells prelabelled with [3H]choline were stimulated with bradykinin there was no enhancement of formation of water soluble products of phosphatidylcholine hydrolysis. When chromaffin cells were permeabilised with pneumolysin and incubated in the presence of [γ‐32P]ATP, the formation of [32P]PA was still stimulated by bradykinin. These results show that, although both neuronal models synthesize PA in response to bradykinin, they do so by quite different routes: PLC/DAG kinase for chromaffin cells and PLD for PC12 cells. The observation that neither bradykinin nor tetradecanoyl phorbol acetate stimulate PLD in chromaffin cells suggests that these cells lack PLD activity. The conservation of PA formation, albeit by different routes, may indicate an essential role of PA in the regulation of cellular events by bradykinin.

Original languageEnglish
Pages (from-to)1084-1087
Number of pages4
JournalJournal of Neurochemistry
Volume57
Issue number3
DOIs
Publication statusPublished - 1 Sep 1991
Externally publishedYes

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