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J. Biol. Chem., Vol. 283, Issue 19, 12949-12959, May 9, 2008

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Linoleic Acid Induces Calcium Signaling, Src Kinase Phosphorylation, and Neurotransmitter Release in Mouse CD36-positive Gustatory Cells^*

Abdelghani El-Yassimi, Aziz Hichami, Philippe Besnard, and Naim Akhtar Khan¹

From the Unité Propre de L'Enseignement Supérieure Lipides and Signalisation Cellulaire Equipe d'Accueil 4183, Faculty of Life Sciences Gabriel, 6, Bd Gabriel, Dijon 21000, France and Ecole Nationale Supérieure de Biologie Appliquéeà la Nutrition et à l'Alimentation, UMR INSERM U866, University of Burgundy, Dijon 21000, France

We have recently demonstrated that the cells expressing CD36, localized apically on the taste buds of mouse lingual circumvallate papillae, act as gustatory cells. In the present study we isolated these CD36-positive cells from mouse circumvallate papillae and investigated intracellular signaling events, triggered by a long-chain polyunsaturated fatty acid, i.e. linoleic acid (LA). LA induced increases in free intracellular calcium concentrations, [Ca²⁺]_i, by recruiting calcium from endoplasmic reticulum pool via inositol 1,4,5-triphosphate production followed by calcium influx via opening of store-operated calcium (SOC) channels. LA also induced phosphorylation of Src-protein-tyrosine kinases (Src-PTKs), particularly of Fyn⁵⁹ and Yes⁶². LA-evoked phosphorylation of Fyn⁵⁹ and Yes⁶² was implicated in the activation of SOC channels. Reverse transcription-quantitative PCR revealed that the CD36-positive gustatory cells possessed mRNA of enzymes like tryptophan hydroxylase-1, L-aromatic amino acid decarboxylase, tyrosine hydroxylase, and dopamine β-hydroxylase, involved in the synthesis of monoamine neurotransmitters. Interestingly, the addition of LA to these cells induced the release of 5-hydroxytryptamine and noradrenalin to the extracellular environment. The LA-induced release of these neurotransmitters was curtailed by SOC channel blockers and Src-PTK inhibitors. These results altogether demonstrate that LA binds to mouse CD36-positive gustatory cells, induces Src-PTKs phosphorylation, triggers calcium signaling, and evokes the release of 5-hydroxytryptamine and noradrenalin, which in turn may be implicated in the downstream signaling to the afferent nerve fibers, thus transmitting the output signal from taste buds to the central nervous system.

Received for publication, September 6, 2007 , and in revised form, February 5, 2008.

^* This work was supported by the National Institute of Agronomic Research, the INSERM through the Research Program in Human Nutrition (to P. B.), and the contingent grants from Agence Nationale pour la Recherche (ANR-07-PNRA-015) (to P. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed. Tel.: 33-3-80-39-63-12; Fax: 33-3-80-39-63-30; E-mail: Naim.Khan{at}u-bourgogne.fr.

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