Lipid Metabolites Enhance Secretion Acting on SNARE Microdomains and Altering the Extent and Kinetics of Single Release Events in Bovine Adrenal Chromaffin Cells

Lipid molecules such as arachidonic acid (AA) and sphingolipid metabolites have been implicated in modulation of neuronal and endocrine secretion. Here we compare the effects of these lipids on secretion from cultured bovine chromaffin cells. First, we demonstrate that exogenous sphingosine and AA interact with the secretory apparatus as confirmed by FRET experiments. Examination of plasma membrane SNARE microdomains and chromaffin granule dynamics using total internal reflection fluorescent microscopy (TIRFM) suggests that sphingosine production promotes granule tethering while arachidonic acid promotes full docking. Our analysis of single granule release kinetics by amperometry demonstrated that both sphingomyelinase and AA treatments enhanced drastically the amount of catecholamines released per individual event by either altering the onset phase of or by prolonging the off phase of single granule catecholamine release kinetics. Together these results demonstrate that the kinetics and extent of the exocytotic fusion pore formation can be modulated by specific signalling lipids through related functional mechanisms.

[1]  Cristina J. Torregrosa-Hetland,et al.  Cortical F-actin affects the localization and dynamics of SNAP-25 membrane clusters in chromaffin cells. , 2013, The international journal of biochemistry & cell biology.

[2]  I. López-Font,et al.  t‐SNARE cluster organization and dynamics in chromaffin cells , 2010, Journal of neurochemistry.

[3]  B. Davletov,et al.  α‐Synuclein sequesters arachidonic acid to modulate SNARE‐mediated exocytosis , 2010, EMBO reports.

[4]  F. Thol,et al.  Erythrocytes serve as a reservoir for cellular and extracellular sphingosine 1‐phosphate , 2010, Journal of cellular biochemistry.

[5]  A. Landar,et al.  Critical Methods in Free Radical Biology & Medicine Methods for imaging and detecting modification of proteins by reactive lipid species , 2009 .

[6]  M. Kreft,et al.  Sphingosine Facilitates SNARE Complex Assembly and Activates Synaptic Vesicle Exocytosis , 2009, Neuron.

[7]  R. Bittman,et al.  Sphingosine 1-phosphate lyase enzyme assay using a BODIPY-labeled substrate. , 2009, Biochemical and biophysical research communications.

[8]  S. Viniegra,et al.  Vesicle Motion and Fusion are Altered in Chromaffin Cells with Increased SNARE Cluster Dynamics , 2009, Traffic.

[9]  Michael J. Saxton,et al.  SINGLE-PARTICLE TRACKING , 2009 .

[10]  T. Lang,et al.  Interplay between lipids and the proteinaceous membrane fusion machinery. , 2008, Progress in lipid research.

[11]  M. Lindau,et al.  The role of the C terminus of the SNARE protein SNAP-25 in fusion pore opening and a model for fusion pore mechanics , 2008, Proceedings of the National Academy of Sciences.

[12]  Yusuf A. Hannun,et al.  Principles of bioactive lipid signalling: lessons from sphingolipids , 2008, Nature Reviews Molecular Cell Biology.

[13]  A. Nakano,et al.  Phospholipid mediated plasticity in exocytosis observed in PC12 cells , 2007, Brain Research.

[14]  B. Davletov,et al.  Regulation of SNARE fusion machinery by fatty acids , 2007, Cellular and Molecular Life Sciences.

[15]  B. Davletov,et al.  Mechanism of arachidonic acid action on syntaxin–Munc18 , 2007, EMBO reports.

[16]  Ute Becherer,et al.  Primed Vesicles Can Be Distinguished from Docked Vesicles by Analyzing Their Mobility , 2007, The Journal of Neuroscience.

[17]  F. Meunier,et al.  Arachidonic acid potentiates exocytosis and allows neuronal SNARE complex to interact with Munc18a , 2006, Journal of neurochemistry.

[18]  J. Kornhuber,et al.  Light‐induced exocytosis in cell development and differentiation , 2006, Journal of cellular biochemistry.

[19]  D. Sulzer,et al.  Analysis of exocytotic events recorded by amperometry , 2005, Nature Methods.

[20]  B. Davletov,et al.  Arachidonic acid allows SNARE complex formation in the presence of Munc18. , 2005, Chemistry & biology.

[21]  A. Villarroel,et al.  New Roles of Myosin II during Vesicle Transport and Fusion in Chromaffin Cells* , 2004, Journal of Biological Chemistry.

[22]  Kees Jalink,et al.  Correcting confocal acquisition to optimize imaging of fluorescence resonance energy transfer by sensitized emission. , 2004, Biophysical journal.

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

[24]  E. Neher,et al.  Differential Control of the Releasable Vesicle Pools by SNAP-25 Splice Variants and SNAP-23 , 2003, Cell.

[25]  M. Criado,et al.  Modifications in the C Terminus of the Synaptosome-associated Protein of 25 kDa (SNAP-25) and in the Complementary Region of Synaptobrevin Affect the Final Steps of Exocytosis* , 2002, The Journal of Biological Chemistry.

[26]  B. Davletov,et al.  Vesicular restriction of synaptobrevin suggests a role for calcium in membrane fusion , 2002, Nature.

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

[28]  M. Criado,et al.  A single amino acid near the C terminus of the synaptosomeassociated protein of 25 kDa (SNAP-25) is essential for exocytosis in chromaffin cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[29]  E. Neher,et al.  Early requirement for α‐SNAP and NSF in the secretory cascade in chromaffin cells , 1999 .

[30]  E. Neher,et al.  Early requirement for alpha-SNAP and NSF in the secretory cascade in chromaffin cells. , 1999, The EMBO journal.

[31]  A. Gil,et al.  Dual effects of botulinum neurotoxin A on the secretory stages of chromaffin cells , 1998, The European journal of neuroscience.

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

[33]  D. Loerke,et al.  The last few milliseconds in the life of a secretory granule , 1998, European Biophysics Journal.

[34]  A. Ferrer-Montiel,et al.  A Peptide That Mimics the C-terminal Sequence of SNAP-25 Inhibits Secretory Vesicle Docking in Chromaffin Cells* , 1997, The Journal of Biological Chemistry.

[35]  S Falkow,et al.  FACS-optimized mutants of the green fluorescent protein (GFP). , 1996, Gene.

[36]  N. Barton,et al.  SNAP-25, a t-SNARE which binds to both syntaxin and synaptobrevin via domains that may form coiled coils. , 1994, The Journal of biological chemistry.

[37]  Wilson Mc,et al.  Human cDNA clones encoding two different isoforms of the nerve terminal protein SNAP-25 , 1994 .

[38]  M. Wilson,et al.  Human cDNA clones encoding two different isoforms of the nerve terminal protein SNAP-25. , 1994, Gene.

[39]  Mark K. Bennett,et al.  A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion , 1993, Cell.

[40]  R. Scheller,et al.  The molecular machinery for secretion is conserved from yeast to neurons. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[41]  J. A. Jankowski,et al.  Temporally resolved catecholamine spikes correspond to single vesicle release from individual chromaffin cells. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[42]  H. Qian,et al.  Single particle tracking. Analysis of diffusion and flow in two-dimensional systems. , 1991, Biophysical journal.

[43]  J. Zimmerberg,et al.  A Lipid/Protein Complex Hypothesis for Exocytotic Fusion Pore Formation a , 1991, Annals of the New York Academy of Sciences.

[44]  P. Zahler,et al.  Arachidonic Acid Liberated by Diacylglycerol Lipase Is Essential for the Release Mechanism in Chromaffin Cells from Bovine Adrenal Medulla , 1990, Journal of neurochemistry.