Vesicular glutamate transporter 3 is required for synaptic transmission in zebrafish hair cells

Nikolaus Obholzer, Sean Wolfson, Josef G. Trapani, Weike Mo, Alex Nechiporuk, Elisabeth Busch-Nentwich, Christoph Seiler, Samuel Sidi, Christian Söllner, Robert N. Duncan, Andrea Boehland, Teresa Nicolson

Research output: Contribution to journalArticle

175 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)2110-2118
Number of pages9
JournalJournal of Neuroscience
Volume28
Issue number9
DOIs
StatePublished - Feb 27 2008

Fingerprint

Vesicular Glutamate Transport Proteins
Minor Planets
Zebrafish
Synaptic Transmission
Synaptic Vesicles
Synaptic Potentials
Startle Reflex
Lateral Line System
Ethylnitrosourea
Neurons
Morpholinos
Mutagenesis
Synapses
Genes
Action Potentials
In Situ Hybridization
Ear
Larva
Glutamic Acid
Exons

Keywords

  • Action currents
  • Hair cells
  • Ribbon synapse
  • Synaptic vesicles
  • Vglut1
  • Vglut3
  • Zebrafish

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Vesicular glutamate transporter 3 is required for synaptic transmission in zebrafish hair cells. / Obholzer, Nikolaus; Wolfson, Sean; Trapani, Josef G.; Mo, Weike; Nechiporuk, Alex; Busch-Nentwich, Elisabeth; Seiler, Christoph; Sidi, Samuel; Söllner, Christian; Duncan, Robert N.; Boehland, Andrea; Nicolson, Teresa.

In: Journal of Neuroscience, Vol. 28, No. 9, 27.02.2008, p. 2110-2118.

Research output: Contribution to journalArticle

Obholzer, N, Wolfson, S, Trapani, JG, Mo, W, Nechiporuk, A, Busch-Nentwich, E, Seiler, C, Sidi, S, Söllner, C, Duncan, RN, Boehland, A & Nicolson, T 2008, 'Vesicular glutamate transporter 3 is required for synaptic transmission in zebrafish hair cells', Journal of Neuroscience, vol. 28, no. 9, pp. 2110-2118. https://doi.org/10.1523/JNEUROSCI.5230-07.2008
Obholzer, Nikolaus ; Wolfson, Sean ; Trapani, Josef G. ; Mo, Weike ; Nechiporuk, Alex ; Busch-Nentwich, Elisabeth ; Seiler, Christoph ; Sidi, Samuel ; Söllner, Christian ; Duncan, Robert N. ; Boehland, Andrea ; Nicolson, Teresa. / Vesicular glutamate transporter 3 is required for synaptic transmission in zebrafish hair cells. In: Journal of Neuroscience. 2008 ; Vol. 28, No. 9. pp. 2110-2118.
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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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AU - Obholzer, Nikolaus

AU - Wolfson, Sean

AU - Trapani, Josef G.

AU - Mo, Weike

AU - Nechiporuk, Alex

AU - Busch-Nentwich, Elisabeth

AU - Seiler, Christoph

AU - Sidi, Samuel

AU - Söllner, Christian

AU - Duncan, Robert N.

AU - Boehland, Andrea

AU - Nicolson, Teresa

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N2 - 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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