Accumulation of an antidepressant in vesiculogenic membranes of yeast cells triggers autophagy

PLoS One. 2012;7(4):e34024. doi: 10.1371/journal.pone.0034024. Epub 2012 Apr 18.

Abstract

Many antidepressants are cationic amphipaths, which spontaneously accumulate in natural or reconstituted membranes in the absence of their specific protein targets. However, the clinical relevance of cellular membrane accumulation by antidepressants in the human brain is unknown and hotly debated. Here we take a novel, evolutionarily informed approach to studying the effects of the selective-serotonin reuptake inhibitor sertraline/Zoloft® on cell physiology in the model eukaryote Saccharomyces cerevisiae (budding yeast), which lacks a serotonin transporter entirely. We biochemically and pharmacologically characterized cellular uptake and subcellular distribution of radiolabeled sertraline, and in parallel performed a quantitative ultrastructural analysis of organellar membrane homeostasis in untreated vs. sertraline-treated cells. These experiments have revealed that sertraline enters yeast cells and then reshapes vesiculogenic membranes by a complex process. Internalization of the neutral species proceeds by simple diffusion, is accelerated by proton motive forces generated by the vacuolar H(+)-ATPase, but is counteracted by energy-dependent xenobiotic efflux pumps. At equilibrium, a small fraction (10-15%) of reprotonated sertraline is soluble while the bulk (90-85%) partitions into organellar membranes by adsorption to interfacial anionic sites or by intercalation into the hydrophobic phase of the bilayer. Asymmetric accumulation of sertraline in vesiculogenic membranes leads to local membrane curvature stresses that trigger an adaptive autophagic response. In mutants with altered clathrin function, this adaptive response is associated with increased lipid droplet formation. Our data not only support the notion of a serotonin transporter-independent component of antidepressant function, but also enable a conceptual framework for characterizing the physiological states associated with chronic but not acute antidepressant administration in a model eukaryote.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antidepressive Agents / metabolism*
  • Antidepressive Agents / pharmacology
  • Autophagy* / drug effects
  • Cell Membrane Permeability
  • Drug Resistance, Fungal / genetics
  • Intracellular Membranes / metabolism*
  • Intracellular Membranes / ultrastructure
  • Kinetics
  • Lysosomes / metabolism
  • Mutation
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Serotonin Plasma Membrane Transport Proteins / genetics
  • Sertraline / metabolism
  • Sertraline / pharmacology

Substances

  • Antidepressive Agents
  • Serotonin Plasma Membrane Transport Proteins
  • Sertraline