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ZIRC
ZFIN ID: ZDB-PUB-080309-24
Vesicular glutamate transporter 3 is required for synaptic transmission in zebrafish hair cells
Obholzer, N., Wolfson, S., Trapani, J.G., Mo, W., Nechiporuk, A., Busch-Nentwich, E., Seiler, C., Sidi, S., Söllner, C., Duncan, R.N., Boehland, A., and Nicolson, T.
Date: 2008
Source: The Journal of neuroscience : the official journal of the Society for Neuroscience   28(9): 2110-2118 (Journal)
Registered Authors: Busch-Nentwich, Elisabeth, Mo, Weike, Nechiporuk, Alex, Nicolson, Teresa, Obholzer, Nikolaus, Seiler, Christoph, Sidi, Samuel, Söllner, Christian, Trapani, Josef, Wolfson, Sean
Keywords: vglut3, zebrafish, action currents, hair cells, synaptic vesicles, ribbon synapse, vglut1
MeSH Terms:
  • Acoustic Stimulation/methods
  • Animals
  • Animals, Genetically Modified
  • Basic Helix-Loop-Helix Transcription Factors/genetics
  • Gene Expression Regulation, Developmental/genetics
  • Gene Expression Regulation, Developmental/physiology
  • Green Fluorescent Proteins/genetics
  • Hair Cells, Auditory/physiology*
  • Larva
  • Microscopy, Electron, Transmission/methods
  • Mutation/physiology
  • Nerve Tissue Proteins/genetics
  • Physical Stimulation/methods
  • Pyridinium Compounds/metabolism
  • Quaternary Ammonium Compounds/metabolism
  • Reflex, Vestibulo-Ocular/physiology
  • Synapses/metabolism
  • Synapses/ultrastructure
  • Synaptic Transmission/physiology*
  • Vesicular Glutamate Transport Proteins/genetics
  • Vesicular Glutamate Transport Proteins/metabolism*
  • Zebrafish
PubMed: 18305245 Full text @ J. Neurosci.
FIGURES
ABSTRACT
Hair cells detect sound and movement and transmit this information via specialized ribbon synapses. Here we report that asteroid, a gene identified in an ethylnitrosourea mutagenesis screen of zebrafish larvae for auditory/vestibular mutants, encodes vesicular glutamate transporter 3 (Vglut3). A splice site mutation in exon 2 of vglut3 results in a severe truncation of the predicted protein product and morpholinos directed against the vglut3 ATG start site or the affected splice junction replicate the asteroid phenotype. In situ hybridization shows that vglut3 is exclusively expressed in hair cells of the ear and lateral line organ. A second transporter gene, vglut1, is also expressed in zebrafish hair cells, but the level of vglut1 mRNA is not increased in the absence of Vglut3. Antibodies against Vglut3 label the basal end of hair cells and labeling is not present in asteroid/vglut3 mutants. Based on the localization of Vglut3 in hair cells, we suspected that the lack of vestibulo-ocular and acoustic startle reflexes in asteroid/vglut3 mutants was attributable to a defect in synaptic transmission in hair cells. In support of this notion, action currents in postsynaptic acousticolateralis neurons are absent in asteroid/vglut3 mutants. At the ultrastructural level, mutant asteroid/vglut3 hair cells show a decrease in the number of ribbon-associated synaptic vesicles, indicating a role for Vglut3 in synaptic vesicle biogenesis and/or tethering to the ribbon body. Lack of postsynaptic action currents in the mutants suggests that the remaining hair-cell synaptic vesicles contain insufficient levels of glutamate for generation of action potentials in first-order neurons.
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