Fife organizes synaptic vesicles and calcium channels for high-probability neurotransmitter release

The strength of synaptic connections varies significantly and is a key determinant of communication within neural circuits. Mechanistic insight into presynaptic factors that establish and modulate neurotransmitter release properties is crucial to understanding synapse strength, circuit function, and neural plasticity. We previously identified Drosophila Piccolo-RIM-related Fife, which regulates neurotransmission and motor behavior through an unknown mechanism. Here, we demonstrate that Fife localizes and interacts with RIM at the active zone cytomatrix to promote neurotransmitter release. Loss of Fife results in the severe disruption of active zone cytomatrix architecture and molecular organization. Through electron tomographic and electrophysiological studies, we find a decrease in the accumulation of release-ready synaptic vesicles and their release probability caused by impaired coupling to Ca2+ channels. Finally, we find that Fife is essential for the homeostatic modulation of neurotransmission. We propose that Fife organizes active zones to create synaptic vesicle release sites within nanometer distance of Ca2+ channel clusters for reliable and modifiable neurotransmitter release.

[1]  H. Zhan,et al.  Three-dimensional imaging of Drosophila motor synapses reveals ultrastructural organizational patterns* , 2016, Journal of neurogenetics.

[2]  Christina A. Beis,et al.  Active zone scaffolds differentially accumulate Unc13 isoforms to tune Ca2+ channel–vesicle coupling , 2016, Nature Neuroscience.

[3]  E. Neher.  Merits and Limitations of Vesicle Pool Models in View of Heterogeneous Populations of Synaptic Vesicles , 2015, Neuron.

[4]  E. F. Stanley.  Single calcium channel domain gating of synaptic vesicle fusion at fast synapses; analysis by graphic modeling , 2015, Channels.

[5]  C. Stigloher,et al.  Presynaptic architecture of the larval zebrafish neuromuscular junction , 2015, The Journal of comparative neurology.

[6]  C. Garner,et al.  Presynaptic active zones in invertebrates and vertebrates , 2015, EMBO reports.

[7]  S. Schoch,et al.  The presynaptic active zone: A dynamic scaffold that regulates synaptic efficacy. , 2015, Experimental cell research.

[8]  H. Zhan,et al.  Advances in imaging ultrastructure yield new insights into presynaptic biology , 2015, Front. Cell. Neurosci..

[9]  G. Davis,et al.  RIM-Binding Protein Links Synaptic Homeostasis to the Stabilization and Replenishment of High Release Probability Vesicles , 2015, Neuron.

[10]  Yukihiro Nakamura,et al.  Ca2+ Channel to Synaptic Vesicle Distance Accounts for the Readily Releasable Pool Kinetics at a Functionally Mature Auditory Synapse , 2015, The Journal of Neuroscience.

[11]  R. Angus Silver,et al.  Nanoscale Distribution of Presynaptic Ca2+ Channels and Its Impact on Vesicular Release during Development , 2015, Neuron.

[12]  T. Südhof,et al.  The Morphological and Molecular Nature of Synaptic Vesicle Priming at Presynaptic Active Zones , 2014, Neuron.

[13]  J. Eilers,et al.  Munc13-3 Superprimes Synaptic Vesicles at Granule Cell-to-Basket Cell Synapses in the Mouse Cerebellum , 2014, The Journal of Neuroscience.

[14]  Uri Ashery,et al.  Quantitative super-resolution imaging of Bruchpilot distinguishes active zone states , 2014, Nature Communications.

[15]  C. Rubinstein,et al.  Highly Specific and Efficient CRISPR/Cas9-Catalyzed Homology-Directed Repair in Drosophila , 2014, Genetics.

[16]  Jeffrey P. Gavornik,et al.  Spontaneous and Evoked Release Are Independently Regulated at Individual Active Zones , 2013, The Journal of Neuroscience.

[17]  W. Ho,et al.  Superpriming of synaptic vesicles after their recruitment to the readily releasable pool , 2013, Proceedings of the National Academy of Sciences.

[18]  Manuela Schmidt,et al.  The Bruchpilot cytomatrix determines the size of the readily releasable pool of synaptic vesicles , 2013, The Journal of cell biology.

[19]  R. Angus Silver,et al.  Sustaining rapid vesicular release at active zones: potential roles for vesicle tethering , 2013, Trends in Neurosciences.

[20]  J. Eilers,et al.  Nanodomain Coupling at an Excitatory Cortical Synapse , 2013, Current Biology.

[21]  Pablo Ariel,et al.  Intrinsic variability in Pv, RRP size, Ca2+ channel repertoire, and presynaptic potentiation in individual synaptic boutons , 2012, Front. Syn. Neurosci..

[22]  Kate M. O’Connor-Giles,et al.  Fife, a Drosophila Piccolo-RIM Homolog, Promotes Active Zone Organization and Neurotransmitter Release , 2012, The Journal of Neuroscience.

[23]  Zhihua Liu,et al.  RIM Promotes Calcium Channel Accumulation at Active Zones of the Drosophila Neuromuscular Junction , 2012, The Journal of Neuroscience.

[24]  G. Davis,et al.  RIM Controls Homeostatic Plasticity through Modulation of the Readily-Releasable Vesicle Pool , 2012, The Journal of Neuroscience.

[25]  R. Tsien,et al.  Synaptic vesicle pools and dynamics. , 2012, Cold Spring Harbor perspectives in biology.

[26]  T. Südhof.  The Presynaptic Active Zone , 2012, Neuron.

[27]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[28]  Masahiko Watanabe,et al.  Release probability of hippocampal glutamatergic terminals scales with the size of the active zone , 2012, Nature Neuroscience.

[29]  D. Sharp,et al.  Transmission electron microscopy of thin sections of Drosophila: high-pressure freezing and freeze-substitution. , 2012, Cold Spring Harbor protocols.

[30]  P. Jonas,et al.  Nanodomain coupling between Ca2+ channels and sensors of exocytosis at fast mammalian synapses , 2011, Nature Reviews Neuroscience.

[31]  Stephan J. Sigrist,et al.  RIM-Binding Protein, a Central Part of the Active Zone, Is Essential for Neurotransmitter Release , 2011, Science.

[32]  P. Verstreken,et al.  ELP3 Controls Active Zone Morphology by Acetylating the ELKS Family Member Bruchpilot , 2011, Neuron.

[33]  P. Simmons,et al.  Structural Organization of the Presynaptic Density at Identified Synapses in the Locust Central Nervous System , 2012 .

[34]  J. Eilers,et al.  Rapid Active Zone Remodeling during Synaptic Plasticity , 2011, The Journal of Neuroscience.

[35]  E. Isacoff,et al.  Optical quantal analysis of synaptic transmission in wild-type and rab3-mutant Drosophila motor axons , 2011, Nature Neuroscience.

[36]  Mei Zhen,et al.  The Presynaptic Dense Projection of the Caenorhabiditis elegans Cholinergic Neuromuscular Junction Localizes Synaptic Vesicles at the Active Zone through SYD-2/Liprin and UNC-10/RIM-Dependent Interactions , 2011, The Journal of Neuroscience.

[37]  Thomas C. Südhof,et al.  RIM Proteins Activate Vesicle Priming by Reversing Autoinhibitory Homodimerization of Munc13 , 2011, Neuron.

[38]  Thomas C. Südhof,et al.  RIM Proteins Tether Ca2+ Channels to Presynaptic Active Zones via a Direct PDZ-Domain Interaction , 2011, Cell.

[39]  O. Shupliakov,et al.  Two pools of vesicles associated with the presynaptic cytosolic projection in Drosophila neuromuscular junctions. , 2010, Journal of structural biology.

[40]  A. Egner,et al.  Bassoon and the Synaptic Ribbon Organize Ca2+ Channels and Vesicles to Add Release Sites and Promote Refilling , 2010, Neuron.

[41]  J. Eilers,et al.  Bassoon Speeds Vesicle Reloading at a Central Excitatory Synapse , 2010, Neuron.

[42]  T. Branco,et al.  Examining size–strength relationships at hippocampal synapses using an ultrastructural measurement of synaptic release probability , 2010, Journal of structural biology.

[43]  E. F. Stanley,et al.  N-type Ca2+ channels carry the largest current: implications for nanodomains and transmitter release , 2010, Nature Neuroscience.

[44]  D. Owald,et al.  Naked Dense Bodies Provoke Depression , 2010, The Journal of Neuroscience.

[45]  J. Richmond,et al.  Differential Regulation of Synaptic Vesicle Tethering and Docking by UNC-18 and TOM-1 , 2010, Front. Syn. Neurosci..

[46]  Jacob Matz,et al.  Rapid structural alterations of the active zone lead to sustained changes in neurotransmitter release , 2010, Proceedings of the National Academy of Sciences.

[47]  S. Hallermann,et al.  Mechanisms of short‐term plasticity at neuromuscular active zones of Drosophila , 2010, HFSP journal.

[48]  V. Lučić,et al.  Quantitative analysis of the native presynaptic cytomatrix by cryoelectron tomography , 2010, The Journal of cell biology.

[49]  Robert W. Burgess,et al.  Rab3 Dynamically Controls Protein Composition at Active Zones , 2009, Neuron.

[50]  Greta Hultqvist,et al.  A Protein Interaction Node at the Neurotransmitter Release Site: Domains of Aczonin/Piccolo, Bassoon, CAST, and Rim Converge on the N-Terminal Domain of Munc13-1 , 2009, The Journal of Neuroscience.

[51]  D. Owald,et al.  Maturation of active zone assembly by Drosophila Bruchpilot , 2009, The Journal of cell biology.

[52]  O. Pascual,et al.  A common molecular basis for membrane docking and functional priming of synaptic vesicles , 2009, The European journal of neuroscience.

[53]  M. Frotscher,et al.  Nanodomain Coupling between Ca2+ Channels and Ca2+ Sensors Promotes Fast and Efficient Transmitter Release at a Cortical GABAergic Synapse , 2008, Neuron.

[54]  S. Schoch,et al.  Structure and evolution of RIM-BP genes: identification of a novel family member. , 2007, Gene.

[55]  Takeshi Sakaba,et al.  The Coupling between Synaptic Vesicles and Ca2+ Channels Determines Fast Neurotransmitter Release , 2007, Neuron.

[56]  E. Réal,et al.  Analysis of synaptic ultrastructure without fixative using high‐pressure freezing and tomography , 2006, The European journal of neuroscience.

[57]  C. A. Frank,et al.  Mechanisms Underlying the Rapid Induction and Sustained Expression of Synaptic Homeostasis , 2006, Neuron.

[58]  Wonhwa Cho,et al.  Membrane binding and subcellular targeting of C2 domains. , 2006, Biochimica et biophysica acta.

[59]  Stephan J. Sigrist,et al.  Bruchpilot Promotes Active Zone Assembly, Ca2+ Channel Clustering, and Vesicle Release , 2006, Science.

[60]  Stephan J. Sigrist,et al.  Bruchpilot, a Protein with Homology to ELKS/CAST, Is Required for Structural Integrity and Function of Synaptic Active Zones in Drosophila , 2006, Neuron.

[61]  T. Südhof,et al.  Rab3 Superprimes Synaptic Vesicles for Release: Implications for Short-Term Synaptic Plasticity , 2006, The Journal of Neuroscience.

[62]  T. Moser,et al.  Few CaV1.3 Channels Regulate the Exocytosis of a Synaptic Vesicle at the Hair Cell Ribbon Synapse , 2005, The Journal of Neuroscience.

[63]  David N Mastronarde,et al.  Automated electron microscope tomography using robust prediction of specimen movements. , 2005, Journal of structural biology.

[64]  Alexander Borst,et al.  Heterogeneity in synaptic transmission along a Drosophila larval motor axon , 2005, Nature Neuroscience.

[65]  E. Neher,et al.  Distinct Kinetic Changes in Neurotransmitter Release After SNARE Protein Cleavage , 2005, Science.

[66]  A. Egner,et al.  Hair cell synaptic ribbons are essential for synchronous auditory signalling , 2005, Nature.

[67]  H. Bellen,et al.  The architecture of the active zone in the presynaptic nerve terminal. , 2004, Physiology.

[68]  R. Tsien,et al.  Presynaptic Ca2+ Channels Compete for Channel Type-Preferring Slots in Altered Neurotransmission Arising from Ca2+ Channelopathy , 2004, Neuron.

[69]  Philippe Rostaing,et al.  Preservation of Immunoreactivity and Fine Structure of Adult C. elegans Tissues Using High-pressure Freezing , 2004, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[70]  Josef Ammermüller,et al.  The Presynaptic Active Zone Protein Bassoon Is Essential for Photoreceptor Ribbon Synapse Formation in the Retina , 2003, Neuron.

[71]  T. Südhof,et al.  Genomic definition of RIM proteins: evolutionary amplification of a family of synaptic regulatory proteins. , 2003, Genomics.

[72]  T. Shibasaki,et al.  Piccolo, a Ca2+ sensor in pancreatic beta-cells. Involvement of cAMP-GEFII.Rim2. Piccolo complex in cAMP-dependent exocytosis. , 2002, The Journal of biological chemistry.

[73]  D. Reiff,et al.  Differential Regulation of Active Zone Density during Long-Term Strengthening of Drosophila Neuromuscular Junctions , 2002, The Journal of Neuroscience.

[74]  F. Kawasaki,et al.  Synaptic Calcium-Channel Function in Drosophila: Analysis and Transformation Rescue of Temperature-Sensitive Paralytic and Lethal Mutations of Cacophony , 2002, The Journal of Neuroscience.

[75]  T. Schikorski,et al.  Inactivity Produces Increases in Neurotransmitter Release and Synapse Size , 2001, Neuron.

[76]  Erik M. Jorgensen,et al.  A post-docking role for active zone protein Rim , 2001, Nature Neuroscience.

[77]  G. Schiavo,et al.  Direct Interaction of the Rab3 Effector RIM with Ca2+Channels, SNAP-25, and Synaptotagmin* , 2001, The Journal of Biological Chemistry.

[78]  Kendal Broadie,et al.  Drosophila Unc-13 is essential for synaptic transmission , 1999, Nature Neuroscience.

[79]  A. C. Meyer,et al.  Released Fraction and Total Size of a Pool of Immediately Available Transmitter Quanta at a Calyx Synapse , 1999, Neuron.

[80]  C. Goodman,et al.  Genetic Analysis of Glutamate Receptors in Drosophila Reveals a Retrograde Signal Regulating Presynaptic Transmitter Release , 1997, Neuron.

[81]  T. Sejnowski,et al.  Heterogeneous Release Properties of Visualized Individual Hippocampal Synapses , 1997, Neuron.

[82]  J. Renger,et al.  Improved stability of Drosophila larval neuromuscular preparations in haemolymph-like physiological solutions , 1994, Journal of Comparative Physiology A.

[83]  Christian Rosenmund,et al.  Nonuniform probability of glutamate release at a hippocampal synapse. , 1993, Science.

[84]  KM Harris,et al.  Dendritic spines of CA 1 pyramidal cells in the rat hippocampus: serial electron microscopy with reference to their biophysical characteristics , 1989, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[85]  T. Reese,et al.  The organization of cytoplasm at the presynaptic active zone of a central nervous system synapse , 1988, Neuron.

[86]  R L Berger,et al.  A stopped-flow investigation of calcium ion binding by ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid. , 1984, Analytical biochemistry.

[87]  P. Jonas,et al.  A small number of open Ca2+ channels trigger transmitter release at a central GABAergic synapse , 2010, Nature Neuroscience.

[88]  W. Betz,et al.  Synaptic vesicle pools , 2005, Nature Reviews Neuroscience.

[89]  A. Hudspeth,et al.  RIM Binding Proteins ( RBPs ) Couple Rab 3-Interacting Molecules ( RIMs ) to Voltage-Gated Ca 2 , 2002 .

[90]  J R Kremer,et al.  Computer visualization of three-dimensional image data using IMOD. , 1996, Journal of structural biology.