Functional Coupling of the NK1 Tachykinin Receptor to Phospholipase D in Chinese Hamster Ovary Cells and Astrocytoma Cells

• Published in: Journal of neurochemistry Vol. 70; no. 5; pp. 2091 - 2098
• Main Authors: Torrens, Yvette, Beaujouan, Jean‐Claude, Saffroy, Monique, Glowinski, Jacques, Tencé, Martine
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01.05.1998; Blackwell
• Subjects: Animals; Astrocytoma - metabolism; Astrocytoma - pathology; Biological and medical sciences; Calcium - pharmacology; Cell receptors; Cell structures and functions; Chinese hamster ovary cells; CHO Cells - metabolism; Cricetinae; Cyclic AMP - physiology; Enzyme Activation - drug effects; Fundamental and applied biological sciences. Psychology; Glycerophospholipids; GTP-Binding Proteins - physiology; Human astrocytoma cells; Humans; Molecular and cellular biology; Myristic Acid - metabolism; Neuropeptide receptors; NK1 receptor; Phosphatidic Acids - biosynthesis; Phosphatidylcholines - metabolism; Phospholipase D; Phospholipase D - metabolism; Protein kinase C; Protein Kinase C - physiology; Rats; Receptors, Neurokinin-1 - metabolism; Substance P; Substance P - analogs & derivatives; Substance P - pharmacology; Tachykinins - pharmacology; Tumor Cells, Cultured; Human; Protein kinase C; NK1 Tachykinin receptor; Calcium; Enzyme; Substance P; Neuroglia; Transferases; Rodentia; Esterases; Phosphoric diester hydrolases; Astrocyte; Neuropeptide; Phospholipase D; Ovary; Signal transduction; Vertebrata; Mammalia; Female genital system; Hydrolases; Tachykinin; Hamster
• ISSN: 0022-3042; 1471-4159
• DOI: 10.1046/j.1471-4159.1998.70052091.x

: In [3H]myristic acid‐prelabeled Chinese hamster ovary cells stably expressing the rat NK1 tachykinin receptor, the selective NK1 agonist [Pro9]substance P ([Pro9]SP) time and concentration dependently stimulated the formation of [3H]phosphatidylethanol in the presence of ethanol. This [Pro9]SP‐induced activation of phospholipase D (PLD) was blocked by NK1 receptor antagonists and poorly or not mimicked by NK2 and NK3 agonists, respectively. In confirmation of previous observations, [Pro9]SP also stimulated the hydrolysis of phosphoinositides, the release of arachidonic acid, and the formation of cyclic AMP (cAMP). All these [Pro9]SP‐evoked responses could be mimicked by aluminum fluoride, but they remained unaffected in cells pretreated with pertussis toxin, suggesting that a Gi/Go protein is not involved in these different signaling pathways. The activation of PLD by [Pro9]SP was sensitive to external calcium and required an active protein kinase C because the inhibition of this kinase (Ro 31‐8220) or its down‐regulation (long‐term treatment with a phorbol ester) abolished the response. In contrast, a cAMP‐dependent process was not involved in the activation of PLD because the [Pro9]SP‐evoked response was neither affected by Rp‐8‐bromoadenosine 3′,5′‐cyclic monophosphorothioate nor mimicked by cAMP‐generating compounds (cholera toxin or forskolin) or by 8‐bromo‐cyclic AMP. A functional coupling of NK1 receptors to PLD was also demonstrated in the human astrocytoma cell line U 373 MG stimulated by SP or [Pro9]SP. These results suggest that PLD activation could be an additional signaling pathway involved in the mechanism of action of SP in target cells expressing NK1 receptors.