Identification and characterization of Drosophila genes for synaptic vesicle proteins

DiAntonio, Aaron; Burgess, RW; Chin, AC; Deitcher, David L.; Scheller, RH; Schwarz, TL

doi:10.1523/jneurosci.13-11-04924.1993

Cited by 143 publications

(96 citation statements)

References 49 publications

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“…By contrast, Sh Kch are abundant in most axonal fibers of the adult brain, whereas only a few synaptic regions appear to have these channels. This nonuniform localization in synaptic areas is supported further by comparison with the distribution patterns of synaptic proteins such as syntaxin (Cerezo et al, 1995), synaptotagmin, and synaptobrevin (DiAntonio et al, 1993). Together, this demonstrates a restricted expression of Sh Kch to certain neuronal populations, as suggested previously from electrical recordings performed in primary cultures (Baker and Salkoff, 1990) and studies performed with Sh transformant flies (Zhao et al, 1995).…”

Section: Diverse Functional Roles Can Be Inferred From the Distributisupporting

confidence: 82%

Diverse Expression and Distribution ofShakerPotassium Channels during the Development of theDrosophilaNervous System

1997

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The spatio-temporal expression ofShaker(Sh) potassium channels (Kch) in the developing and adult nervous system ofDrosophilahas been studied at the molecular and histological level using specific antisera.ShKch are distributed in most regions of the nervous system, but their expression is restricted to only certain populations of cells.ShKch have been found in the following three locations: in synaptic areas of neuropile, in axonal fiber tracks, and in a small number of neuronal cell bodies. This wide subcellular localization, together with a diverse distribution, implicatesShKch in multiple neuronal functions.Experiments performed withShmutants that specifically eliminate a few of theShKch splice variants clearly demonstrate an abundant differential expression and usage of the wide repertoire ofShisoforms, but they do not support the idea of extensive segregation of these isoforms among different populations of neurons.ShKch are predominantly expressed at late stages of postembryonic development and adulthood. Strikingly, wide changes in the repertoire ofShsplice isoforms occur some time after the architecture of the nervous system is complete, indicating that the expression ofShKch contributes to the final refinements of neuronal differentiation. These late changes in the expression and distribution ofShKch seem to correlate with activity patterns suggesting thatShKch may be involved in adaptative mechanisms of excitability.

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Section: Diverse Functional Roles Can Be Inferred From the Distributisupporting

confidence: 82%

Diverse Expression and Distribution ofShakerPotassium Channels during the Development of theDrosophilaNervous System

1997

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“…The hydrophobic transmembrane anchor (boxed) is followed by a long intravesicular domain. Similar (but not homologous) domains also are found in other invertebrate synaptobrevins such as those of Drosophila and Aplysia, but not in mammalian synaptobrevins (Südhof et al, 1989;DiAntonio et al, 1993;Yamasaki et al, 1994). (Elferink et al, 1989), Torpedo (Trimble at al., 1988), Loligo (Hunt et al, 1994), Drosophila (DiAntonio et al, 1993), and Aplysia .…”

Section: Leech Synaptobrevin and Snap-25 Are Homologous To Their Mammmentioning

confidence: 74%

“…Leech synaptobrevin is highly homologous to isoforms from other invertebrates and vertebrates (Elferink et al, 1989;Südhof et al, 1989;Archer et al, 1990;DiAntonio et al, 1993;Hunt et al, 1994;Yamasaki et al, 1994), including conservation of the toxin cleavage site. Like its mammalian homolog (Söllner et al, 1993b;Hayashi et al, 1994) Hirudo synaptobrevin is able to form a stable ternary complex with the leech isoforms of SNAP-25 and syntaxin.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Transmitter Release Correlates with the Proteolytic Activity of Tetanus Toxin and Botulinus Toxin A in Individual Cultured Synapses ofHirudo medicinalis

et al. 1997

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We have studied the effects of tetanus toxin and botulinus toxin A on neurotransmitter release in the Retzius3 P-cell synapse of the leech and exploited the unique properties of this system, which allow for combined physiological and biochemical analyses in single-cell pairs. The sequences of Hirudo medicinalis synaptobrevin and synaptosomal-associated protein of 25 kDa , deduced by cDNA cloning, are 61 and 55% identical, respectively, to their corresponding mammalian homologs. Whereas Hirudo synaptobrevin is proteolyzed by tetanus toxin, its SNAP-25 isoform is resistant to botulinus toxin A cleavage because of amino acid substitutions within and around the putative cleavage site. In close correlation, microinjection of tetanus toxin into the presynaptic neuron produced a block of transmitter release, whereas botulinus toxin A had no effect on synaptic transmission. Subsequent immunoblotting of single-cell pairs demonstrated directly that the tetanus toxinmediated block of exocytosis is accompanied by cleavage of synaptobrevin in the injected neuron, resulting in the generation of a detectable C-terminal cleavage product. Immunoblotting also confirmed the resistance of SNAP-25 to botulinus toxin A cleavage in vivo. Using recombinant proteins, we show that the N-terminal fragment of synaptobrevin released by tetanus toxin, but not its C-terminal membrane-anchored cleavage product, participates with syntaxin and SNAP-25 in synaptic SNAP receptor (SNARE) ternary complex formation in Hirudo. Our data demonstrate a direct correlation between the inhibition of transmitter release and the ability of the neurotoxin to proteolyze its target protein and support the view that SNARE ternary complex formation is an important step leading to synaptic vesicle exocytosis. Key words: transmitter release; synaptobrevin; VAMP; SNAP-25; SNARE; tetanus toxin; botulinus toxin A; single-cell immunoblotting; Hirudo medicinalisCalcium-regulated exocytosis of synaptic vesicles is the primary means of communication between neurons. A complex of conserved cytosolic proteins known as NSF (N-ethylmaleimidesensitive fusion protein) and SNAP proteins (soluble NSF attachment proteins) is required for various intracellular fusion events in eukaryotic cells (Ferro-Novick and Jahn, 1994;Südhof, 1995;Calakos and Scheller, 1996;Rothman and Wieland, 1996). The search for neuronal membrane receptors (SNAREs, SNAP receptors) for these soluble "fusion" factors led to the identification of the vesicle protein synaptobrevin (VAMP) and the plasma membrane proteins, syntaxin and SNAP-25 (synaptosomal-associated protein of 25 kDa) (Söllner et al., 1993a).Synaptobrevins are class II integral membrane proteins that expose most of their sequence at the cytoplasmic face of secretory vesicles (Südhof, 1995). SNAP-25 is an abundant highly conserved membrane-associated protein (Oyler et al., 1989;Risinger et al., 1993). It is post-translationally modified by palmitoyl side chains that mediate membrane binding (Hess et al., 1992;Veit et al., 1996).Syntaxin is a simi...

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“…Thus, either dDAB does not participate in dSYT1 sorting or sufficient redundancy is provided by interactions of SYT1 with at least two other CLATHRIN-associated adaptor proteins, AP-2 and Stonins (4,36). Furthermore, the distributions of other synaptic vesicle proteins at dab mutant synapses, including neuronal SYN-APTOBREVIN (38) and the vesicular glutamate transporter (39), were similar to those of WT (Fig. S6 K-T and U-D′, respectively).…”

Section: Discussionmentioning

confidence: 90%

The DISABLED protein functions in CLATHRIN-mediated synaptic vesicle endocytosis and exoendocytic coupling at the active zone

Kawasaki

Iyer

Posey

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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Members of the DISABLED (DAB) family of proteins are known to play a conserved role in endocytic trafficking of cell surface receptors by functioning as monomeric CLATHRIN-associated sorting proteins that recruit cargo proteins into endocytic vesicles. Here, we report a Drosophila disabled mutant revealing a novel role for DAB proteins in chemical synaptic transmission. This mutant exhibits impaired synaptic function, including a rapid activitydependent reduction in neurotransmitter release and disruption of synaptic vesicle endocytosis. In presynaptic boutons, Drosophila DAB and CLATHRIN were highly colocalized within two distinct classes of puncta, including relatively dim puncta that were located at active zones and may reflect endocytic mechanisms operating at neurotransmitter release sites. Finally, broader analysis of endocytic proteins, including DYNAMIN, supported a general role for CLATHRIN-mediated endocytic mechanisms in rapid clearance of neurotransmitter release sites for subsequent vesicle priming and refilling of the release-ready vesicle pool.I ntensive study of chemical synaptic transmission has led to detailed molecular models of synaptic vesicle endocytosis (1-4). Ongoing efforts seek to define the underlying molecular components and interactions further as well as their spatial organization with respect to neurotransmitter release sites. The present study reveals novel molecular mechanisms and interactions in synaptic vesicle endocytosis involving a member of the DISABLED family of Phosphotyrosine Binding (PTB) domain proteins. In Drosophila, the disabled (dab) gene, the founding member of the dab gene family, was first identified in a screen for genetic modifiers of abelson (5). Although putative dab mutations were subsequently mapped to a different gene (6), recent studies have established functional interactions of Drosophila DISABLED (dDAB) and ABELSON (7). Proteins closely related to dDAB, including mouse DAB-2 (mDAB-2) and the single Caenorhabditis elegans family member ceDAB-1, function as CLATHRIN-associated adaptors in endocytic trafficking of cell surface receptors (8-10). These DABs have been shown to interact with CLATHRIN, the α-ADAPTIN (AP-2) adaptor complex, and other components of the endocytic machinery through conserved binding motifs (8-10) (Fig. 1A). Notably, mDAB-2, ceDAB-1, and dDAB share a PTB/DAB Homology (DH) domain near the N terminus (Fig. 1B), which interacts with specific vesicle cargo proteins and phosphatidylinositol-4,5-diphosphate (11, 12). Neither DAB proteins nor PTB domain interactions have been previously implicated in synaptic vesicle trafficking.The present study reveals a role for DAB proteins in synaptic vesicle endocytosis and extends previous work on interactions of endocytic and exocytic mechanisms in neurotransmitter release. Here, we use a glutamatergic neuromuscular synapse of the Drosophila adult as a model for genetic analysis of molecular mechanisms determining conserved properties of glutamatergic synapse function (13,14). Previous analysis i...

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Identification and characterization of Drosophila genes for synaptic vesicle proteins

Cited by 143 publications

References 49 publications

Diverse Expression and Distribution ofShakerPotassium Channels during the Development of theDrosophilaNervous System

Diverse Expression and Distribution ofShakerPotassium Channels during the Development of theDrosophilaNervous System

Inhibition of Transmitter Release Correlates with the Proteolytic Activity of Tetanus Toxin and Botulinus Toxin A in Individual Cultured Synapses ofHirudo medicinalis

The DISABLED protein functions in CLATHRIN-mediated synaptic vesicle endocytosis and exoendocytic coupling at the active zone

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