Sensory SystemResearch PaperHeterogeneity of glutamatergic and GABAergic release machinery in cerebral cortex: analysis of synaptogyrin, vesicle-associated membrane protein, and syntaxin
Section snippets
Animals and tissue preparation
Adult male Sprague–Dawley rats (190–220 g; Charles River, Milan, Italy) were used. All experiments were carried out in accordance with the European Community Council Directive dated November 24, 1986 (86/609 EEC) and were approved by the local authority veterinary service. All efforts were made to minimize animal suffering and the number of animals used. Animals were kept under a dark-light cycle of 12 h and permitted food and water ad libitum.
For Western blotting, rats were anesthetized with
Preliminary studies
In cortical crude membrane fractions, all antibodies recognized bands of predicted molecular mass (Fig. 2;Belizaire et al., 2004, Baumert et al., 1989, Baumert et al., 1990, Archer et al., 1990, Ruiz-Montasell et al., 1996, Bennett et al., 1992, Bellocchio et al., 1998, Chaudhry et al., 1998, Varoqui et al., 2002). The distribution of VGLUT1, VGLUT2, VGAT, SGYR1, SGYR3, VAMP1, VAMP2, STX1A and STX1B immunoreactivities was as previously described (Belizaire et al., 2004, Ruiz-Montasell et al.,
Discussion
The present studies showed that diverse proteins participating in neurotransmitter release are differentially expressed in glutamatergic and GABAergic terminals. In addition, they also showed that in terminals expressing a given protein participating in transmitter release, the percentage of synaptic vesicles expressing or interacting with that protein is variable.
Previous localization studies have shown that all VGAT+ puncta (e.g., Chaudhry et al., 1998, Minelli et al., 2003) and the vast
Acknowledgments
This work was supported by grants from Ministero Istruzione Università e Ricerca (PRIN) to FC and FB, and from Compagnia di San Paolo (2005.1964) and Telethon-Italy to FB. We thank R. Janz (University of Texas Medical School at Houston) for synaptogyrin antibodies, and M. Melone and G. Fattorini for help and insightful comments.
References (66)
- et al.
Perisomatic glutamatergic axon terminals: a novel feature of cortical synaptology revealed by vesicular glutamate transporter 1 immunostaining
Neuroscience
(2004) - et al.
Structures and chromosomal localizations of two human genes encoding synaptobrevins 1 and 2
J Biol Chem
(1990) - et al.
Heterogeneity of glutamatergic and GABAergic release machineries in cerebral cortex
Neuroscience
(2007) - et al.
GABA and glycine in synaptic vesicles: storage and transport characteristics
Neuron
(1991) - et al.
Generating diversity at GABAergic synapses
Trends Neurosci
(2001) - et al.
GABA transporters in the mammalian cerebral cortex: localization, development and pathological implications
Brain Res Brain Res Rev
(2004) - et al.
Shaping excitation at glutamatergic synapses
Trends Neurosci
(1999) - et al.
SNAP-25/syntaxin 1A complex functionally modulates neurotransmitter gamma-aminobutyric acid reuptake
J Biol Chem
(2006) - et al.
Syntaxin 1A is required for normal in utero development
Biochem Biophys Res Commun
(2008) - et al.
Postnatal development of the vesicular GABA transporter in rat cerebral cortex
Neuroscience
(2003)
Expression and functional role of syntaxin 1/HPC-1 in pancreatic beta cellsSyntaxin 1A, but not 1B, plays a negative role in regulatory insulin release pathway
J Biol Chem
Munc 18a binding to syntaxin 1A and 1B isoforms defines its localization at the plasma membrane and blocks SNARE assembly in a three-hybrid system assay
Mol Cell Neurosci
Distribution of synaptobrevin/VAMP 1 and 2 in rat brain
J Chem Neuroanat
An isolated pool of vesicles recycles at rest and drives spontaneous neurotransmission
Neuron
Cellular and subcellular localization of syntaxin-like immunoreactivity in the rat striatum and cortex
Neuroscience
Molecular anatomy of a trafficking organelle
Cell
SNAP-25 modulation of calcium dynamics underlies differences in GABAergic and glutamatergic responsiveness to depolarization
Neuron
Munc13-1 is essential for fusion competence of glutamatergic synaptic vesicles
Nature
Synaptobrevin: an integral membrane protein of 18,000 daltons present in small synaptic vesicles of rat brain
EMBO J
P29: a novel tyrosine-phosphorylated membrane protein present in small clear vesicles of neurons and endocrine cells
J Cell Biol
Protein kinase C regulates the interaction between a GABA transporter and syntaxin 1A
J Neurosci
Characterization of synaptogyrin 3 as a new synaptic vesicle protein
J Comp Neurol
The localization of the brain-specific inorganic phosphate transporter suggests a specific presynaptic role in glutamatergic transmission
J Neurosci
Syntaxin: a synaptic protein implicated in docking of synaptic vesicles at presynaptic active zones
Science
Astrocytes contain a vesicular compartment that is competent for regulated exocytosis of glutamate
Nat Neurosci
GAT-1 regulates both tonic and phasic GABAA receptor-mediated inhibition in the cerebral cortex
J Neurochem
Clozapine up-regulates synaptophysin expression in rat frontal cortex
J Neurochem
Sustained neurotransmitter release: new molecular clues
Eur J Neurosci
The vesicular GABA transporter, VGAT, localizes to synaptic vesicles in sets of glycinergic as well as GABAergic neurons
J Neurosci
Heterogeneity of axon terminals expressing VGLUT1 in the cerebral neocortex
Arch Ital Biol
Molecular heterogeneity of central synapses: afferent and target regulation
Nat Neurosci
Localization of the GLYT1 glycine transporter at glutamatergic synapses in the rat brain
Cereb Cortex
Transport rates of GABA transporters: regulation by the N-terminal domain and syntaxin 1A
Nat Neurosci
Cited by (37)
Interneuronal exchange and functional integration of synaptobrevin via extracellular vesicles
2021, NeuronCitation Excerpt :However, syb2-pHluorin from EVs is equally incorporated by glutamatergic and GABAergic presynaptic terminals, indicating that another factor underlies the selectivity of effects. It was previously proposed for cerebellar and cortical neurons that syb2, along with other synaptic vesicle proteins, may be expressed at lower levels in GABAergic presynaptic boutons in comparison to glutamatergic counterparts (Benagiano et al., 2011; Bragina et al., 2010; but also see Grønborg et al., 2010). Therefore, a ceiling effect in excitatory synapses, where the endogenous high levels of syb2 occlude the effects of EVs, may underlie this discrepancy.
Seizing the moment: Zebrafish epilepsy models
2020, Neuroscience and Biobehavioral ReviewsThe Control of Neuronal Calcium Homeostasis by SNAP-25 and its Impact on Neurotransmitter Release
2019, NeuroscienceCitation Excerpt :The different SNARE-dependent modulation of VGCCs was also brought in to explain the tight coupling between the Ca2+ influx and transmitter release at basket cell (BC)-granule cell (GC) synapse in the dentate gyrus (Bucurenciu et al., 2008) and the differential regulation of spontaneous vesicle fusion in cholinergic and GABAergic synapses (Liu et al., 2018). Together, these data support the view that the lower levels of SNAP-25, (and even possibly Syntaxin (Bragina et al., 2010), in inhibitory neurons, through the differential control of VGCCs in the two neuron populations, is at the basis of the distinctive properties of excitatory versus inhibitory neurotransmission. Mutations in the Snap-25 gene have been identified in different neurological and psychiatric disorders such as attention deficit/hyperactivity disorder (ADHD), schizophrenia (Barr et al., 2000; Brophy et al., 2002; Kustanovich et al., 2003) and bipolar disorder (Etain et al., 2010; Houenou et al., 2017).
Synaptic vesicle protein 2: A multi-faceted regulator of secretion
2019, Seminars in Cell and Developmental Biology