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Permissive role of sphingosine on calcium-dependent endocytosis in chromaffin cells

  • Signaling and cell physiology
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Abstract

Sphingosine has been shown to modulate neurotransmitter release. Because membrane fusion and fission involve lipid metabolism, we asked here whether sphingosine had a role in regulating endocytosis. To explore this hypothesis, we monitored changes of membrane capacitance (Cm) to study the effects of intracellular sphingosine on membrane retrieval after chromaffin cell stimulation with depolarising pulses (DPs). We found that: (1) sphingosine dialysis through the patch-clamp pipette (SpD) using the whole-cell configuration of the patch-clamp technique (WCC) favours the appearance of a pronounced endocytotic response; (2) SpD-elicited endocytosis was Ca2+-dependent but Ba2+ did not substitute Ca2+; (3) under WCC, such endocytotic response disappeared with repetitive DPs; (4) in cells preincubated with sphingomyelinase to augment endogenous sphingosine synthesis, and then voltage-clamped under the perforated-patch configuration of the patch-clamp technique (PPC), endocytosis decayed little with repeated stimulation; (5) sphingosine-1-phosphate (S1P), a metabolite of sphingosine, had a meagre effect on endocytosis; and (6) neither dynamin inhibitor dynasore nor calmodulin blocker calmidazolium affected the sphingosine elicited endocytosis. We believe this is the first report showing that sphingosine plays a permissive role in activating Ca2+-dependent endocytosis during cell depolarisation. This effect requires high subplasmalemmal cytosolic Ca2+ concentrations and a cytosolic factor(s) that is dialysed with the pipette solution. Independence of dynamin and calmodulin suggests that sphingosine-dependent endocytosis could be a novel, more direct pathway for vesicle recycling under mild depolarisation stimuli.

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Acknowledgements

This work was partially supported from the following grants from Spanish institutions to AGG: (1) SAF2006-03589, Ministerio de Ciencia e Innovación, Spain; (2) NDE 07/09, Agencia Laín Entralgo, Comunidad de Madrid; (3) PI016/09, Fundación C.I.E.N., Instituto de Salud Carlos III; (4) RD 06/0026 RETICS, Instituto de Salud Carlos III; (5) S-SAL-0275-2006, Comunidad de Madrid, also by grant SAF2007-65181, Ministerio de Ciencia e Innovación, Spain, to LG. We thank Fundación Teófilo Hernando for continued support.

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Authors and Affiliations

  1. Instituto Teófilo Hernando Facultad de Medicina, Universidad Autónoma de Madrid, Arzobispo Morcillo, 4, 28029, Madrid, Spain

    Juliana M. Rosa, Luis Gandía & Antonio G. García

  2. Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain

    Juliana M. Rosa, Luis Gandía & Antonio G. García

  3. Servicio de Farmacología Clínica, Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain

    Antonio G. García

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  1. Juliana M. Rosa
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Correspondence to Antonio G. García.

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Rosa, J.M., Gandía, L. & García, A.G. Permissive role of sphingosine on calcium-dependent endocytosis in chromaffin cells. Pflugers Arch - Eur J Physiol 460, 901–914 (2010). https://doi.org/10.1007/s00424-010-0861-x

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  • DOI: https://doi.org/10.1007/s00424-010-0861-x

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