Regulation of Exocytosis in Chromaffin Cells by Trans‐Insertion of Lysophosphatidylcholine and Arachidonic Acid into the Outer Leaflet of the Cell Membrane

Vesicular exocytosis is an important complex process in the communication between cells in organisms. It controls the release of chemical and biochemical messengers stored in an emitting cell. In this report, exocytosis is studied amperometrically (at carbon fiber ultramicroelectrodes) at adrenal chromaffin cells, which release catecholamines after appropriate stimulation, while testing the effects due to trans‐insertion of two exogenous compounds (lysophosphatidylcholine (LPC) and arachidonic acid (AA)) on the kinetics of exocytotic events. Amperometric analyses showed that, under the present conditions (short incubation times and micromolar LPC or AA solutions), LPC favors catecholamine release (rate, event frequency, charge released) while AA disfavors the exocytotic processes. The observed kinetic features are rationalized quantitatively by considering a stalk model, for the fusion pore formation, and the physical constraints applied to the cell membrane by the presence of small fractions of LPC and AA diluted in its external leaflet (trans‐insertion). We also observed that the detected amount of neurotransmitters in the presence of LPC was larger than under control conditions, while the opposite trend is observed with AA.

[1]  A. Ewing,et al.  Correlation between vesicle quantal size and fusion pore release in chromaffin cell exocytosis. , 2005, Biophysical journal.

[2]  T. McIntosh,et al.  Melittin-induced bilayer leakage depends on lipid material properties: evidence for toroidal pores. , 2005, Biophysical journal.

[3]  K. Stiasny,et al.  Effect of Membrane Curvature-Modifying Lipids on Membrane Fusion by Tick-Borne Encephalitis Virus , 2004, Journal of Virology.

[4]  A. Ewing,et al.  The Effects of Vesicular Volume on Secretion through the Fusion Pore in Exocytotic Release from PC12 Cells , 2004, The Journal of Neuroscience.

[5]  M. Kozlov,et al.  Protein-lipid interplay in fusion and fission of biological membranes. , 2003, Annual review of biochemistry.

[6]  Alan Morgan,et al.  Secretory granule exocytosis. , 2003, Physiological reviews.

[7]  M. Erard,et al.  Dynamics of full fusion during vesicular exocytotic events: release of adrenaline by chromaffin cells. , 2003, Chemphyschem : a European journal of chemical physics and physical chemistry.

[8]  J. Barclay,et al.  Splitting the quantum: regulation of quantal release during vesicle fusion , 2002, Trends in Neurosciences.

[9]  Yonathan Kozlovsky,et al.  Stalk model of membrane fusion: solution of energy crisis. , 2002, Biophysical journal.

[10]  H. Palfrey,et al.  Quantal Size Is Dependent on Stimulation Frequency and Calcium Entry in Calf Chromaffin Cells , 2001, Neuron.

[11]  R. Rand,et al.  The influence of lysolipids on the spontaneous curvature and bending elasticity of phospholipid membranes. , 2001, Biophysical journal.

[12]  Daniel Axelrod,et al.  Restriction of Secretory Granule Motion near the Plasma Membrane of Chromaffin Cells , 2001, The Journal of cell biology.

[13]  Richard H. Scheller,et al.  SNARE-mediated membrane fusion , 2001, Nature Reviews Molecular Cell Biology.

[14]  Misuzu Baba,et al.  Geranylgeranylated Snares Are Dominant Inhibitors of Membrane Fusion , 2000, The Journal of cell biology.

[15]  J. Rothman,et al.  Compartmental specificity of cellular membrane fusion encoded in SNARE proteins , 2000, Nature.

[16]  S. Chasserot-Golaz,et al.  Regulation of exocytosis in chromaffin cells by phosducin‐like protein, a protein interacting with G protein βγ subunits , 2000 .

[17]  D. Zenisek,et al.  Transport, capture and exocytosis of single synaptic vesicles at active zones , 2000, Nature.

[18]  R. Wightman,et al.  Adrenaline Release by Chromaffin Cells: Constrained Swelling of the Vesicle Matrix Leads to Full Fusion , 2000 .

[19]  R. Wightman,et al.  Interplay between membrane dynamics, diffusion and swelling pressure governs individual vesicular exocytotic events during release of adrenaline by chromaffin cells. , 2000, Biochimie.

[20]  Y. Humeau,et al.  How botulinum and tetanus neurotoxins block neurotransmitter release. , 2000, Biochimie.

[21]  J. Zimmerberg,et al.  Dynamics of fusion pores connecting membranes of different tensions. , 2000, Biophysical journal.

[22]  C. Amatore,et al.  Time-Resolved Dynamics of the Vesicle Membrane During Individual Exocytotic Secretion Events, as Extracted from Amperometric Monitoring of Adrenaline Exocytosis from Chromaffin Cells , 1999 .

[23]  T. McIntosh,et al.  Membrane fusion promoters and inhibitors have contrasting effects on lipid bilayer structure and undulations. , 1999, Biophysical journal.

[24]  P. Ascher,et al.  Opposite modulation of NMDA receptors by lysophospholipids and arachidonic acid: common features with mechanosensitivity , 1998, The Journal of physiology.

[25]  Reinhard Jahn,et al.  Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 Å resolution , 1998, Nature.

[26]  Benedikt Westermann,et al.  SNAREpins: Minimal Machinery for Membrane Fusion , 1998, Cell.

[27]  R. Schlegel,et al.  A Subfamily of P-Type ATPases with Aminophospholipid Transporting Activity , 1996, Science.

[28]  J. A. Jankowski,et al.  Monitoring an oxidative stress mechanism at a single human fibroblast. , 1995, Analytical chemistry.

[29]  J. Zimmerberg,et al.  The hemifusion intermediate and its conversion to complete fusion: regulation by membrane composition. , 1995, Biophysical journal.

[30]  J. M. Fernández,et al.  Release of secretory products during transient vesicle fusion , 1993, Nature.

[31]  Steven S. Vogel,et al.  Lysolipids reversibly inhibit Ca2+‐, GTP‐ and pH‐dependent fusion of biological membranes , 1993, FEBS letters.

[32]  D. Leckband,et al.  Role of calcium in the adhesion and fusion of bilayers. , 1993, Biochemistry.

[33]  J. A. Jankowski,et al.  Analysis of diffusional broadening of vesicular packets of catecholamines released from biological cells during exocytosis. , 1992, Analytical chemistry.

[34]  B. Malewicz,et al.  Lysophosphatidylcholine stabilizes small unilamellar phosphatidylcholine vesicles. Phosphorus-31 NMR evidence for the "wedge" effect. , 1989, Biophysical journal.

[35]  E. W. Thompson,et al.  Asymmetry of lysophosphatidylcholine/cholesterol vesicles is sensitive to cholesterol modulation. , 1988, Biochemistry.

[36]  G. Giacometti,et al.  Isocyanide binding kinetics to monomeric hemoproteins. A study on the ligand partition between solvent and heme pocket. , 1987, Biophysical journal.

[37]  B. Livett Adrenal medullary chromaffin cells in vitro. , 1984, Physiological reviews.

[38]  P. Cullis,et al.  The role of nonbilayer lipid structures in the fusion of human erythrocytes induced by lipid fusogens. , 1981, Biochimica et biophysica acta.

[39]  J. K. Thomas,et al.  Interaction of lysophosphatidylcholine with phosphatidylcholine bilayers. A photo-physical and NMR study. , 1980, Biochimica et biophysica acta.

[40]  B. Ninham,et al.  Theory of self-assembly of hydrocarbon amphiphiles into micelles and bilayers , 1976 .