Abstract
Three subtypes of vesicular transporters accumulate glutamate into synaptic vesicles to promote its vesicular release. One of the subtypes, VGLUT3, is expressed in neurons, including cholinergic striatal interneurons, that are known to release other classical transmitters. Here we showed that disruption of the Slc17a8 gene (also known as Vglut3) caused an unexpected hypocholinergic striatal phenotype. Vglut3−/− mice were more responsive to cocaine and less prone to haloperidol-induced catalepsy than wild-type littermates, and acetylcholine release was decreased in striatum slices lacking VGLUT3. These phenotypes were associated with a colocalization of VGLUT3 and the vesicular acetylcholine transporter (VAChT) in striatal synaptic vesicles and the loss of a synergistic effect of glutamate on vesicular acetylcholine uptake. We propose that this vesicular synergy between two transmitters is the result of the unbalanced bioenergetics of VAChT, which requires anion co-entry for continuing vesicular filling. Our study reveals a previously unknown effect of glutamate on cholinergic synapses with potential functional and pharmacological implications.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Fremeau, R.T., Jr, Voglmaier, S., Seal, R.P. & Edwards, R.H. VGLUTs define subsets of excitatory neurons and suggest novel roles for glutamate. Trends Neurosci. 27, 98–103 (2004).
Fremeau, R.T., Jr et al. The identification of vesicular glutamate transporter 3 suggests novel modes of signaling by glutamate. Proc. Natl. Acad. Sci. USA 99, 14488–14493 (2002).
Gras, C. et al. A third vesicular glutamate transporter expressed by cholinergic and serotoninergic neurons. J. Neurosci. 22, 5442–5451 (2002).
Schafer, M.K., Varoqui, H., Defamie, N., Weihe, E. & Erickson, J.D. Molecular cloning and functional identification of mouse vesicular glutamate transporter 3 and its expression in subsets of novel excitatory neurons. J. Biol. Chem. 277, 50734–50748 (2002).
Takamori, S., Malherbe, P., Broger, C. & Jahn, R. Molecular cloning and functional characterization of human vesicular glutamate transporter 3. EMBO Rep. 3, 798–803 (2002).
Nickerson Poulin, A., Guerci, A., El Mestikawy, S. & Semba, K. Vesicular glutamate transporter 3 immunoreactivity is present in cholinergic basal forebrain neurons projecting to the basolateral amygdala in rat. J. Comp. Neurol. 498, 690–711 (2006).
Herzog, E. et al. Localization of VGLUT3, the vesicular glutamate transporter type 3, in the rat brain. Neuroscience 123, 983–1002 (2004).
Somogyi, J. et al. GABAergic basket cells expressing cholecystokinin contain vesicular glutamate transporter type 3 (VGLUT3) in their synaptic terminals in hippocampus and isocortex of the rat. Eur. J. Neurosci. 19, 552–569 (2004).
Kawaguchi, Y. Large aspiny cells in the matrix of the rat neostriatum in vitro: physiological identification, relation to the compartments and excitatory postsynaptic currents. J. Neurophysiol. 67, 1669–1682 (1992).
Calabresi, P., Centonze, D., Gubellini, P., Pisani, A. & Bernardi, G. Acetylcholine-mediated modulation of striatal function. Trends Neurosci. 23, 120–126 (2000).
Zhou, F.M., Wilson, C.J. & Dani, J.A. Cholinergic interneuron characteristics and nicotinic properties in the striatum. J. Neurobiol. 53, 590–605 (2002).
Graybiel, A.M., Aosaki, T., Flaherty, A.W. & Kimura, M. The basal ganglia and adaptive motor control. Science 265, 1826–1831 (1994).
Hikida, T. et al. Increased sensitivity to cocaine by cholinergic cell ablation in nucleus accumbens. Proc. Natl. Acad. Sci. USA 98, 13351–13354 (2001).
Kaneko, S. et al. Synaptic integration mediated by striatal cholinergic interneurons in basal ganglia function. Science 289, 633–637 (2000).
Kitabatake, Y., Hikida, T., Watanabe, D., Pastan, I. & Nakanishi, S. Impairment of reward-related learning by cholinergic cell ablation in the striatum. Proc. Natl. Acad. Sci. USA 100, 7965–7970 (2003).
Herbin, M., Gasc, J.P. & Renous, S. Symmetrical and asymmetrical gaits in the mouse: patterns to increase velocity. J. Comp. Physiol. A. Neuroethol. Sens. Neural. Behav. Physiol. 190, 895–906 (2004).
Hitzemann, R., Qian, Y. & Hitzemann, B. Dopamine and acetylcholine cell density in the neuroleptic responsive (NR) and neuroleptic nonresponsive (NNR) lines of mice. J. Pharmacol. Exp. Ther. 266, 431–438 (1993).
Hikida, T., Kitabatake, Y., Pastan, I. & Nakanishi, S. Acetylcholine enhancement in the nucleus accumbens prevents addictive behaviors of cocaine and morphine. Proc. Natl. Acad. Sci. USA 100, 6169–6173 (2003).
Tzavara, E.T. et al. Procholinergic and memory enhancing properties of the selective norepinephrine uptake inhibitor atomoxetine. Mol. Psychiatry 11, 187–195 (2006).
Roseth, S., Fykse, E.M. & Fonnum, F. Uptake of l-glutamate into synaptic vesicles: competitive inhibition by dyes with biphenyl and amino- and sulphonic acid–substituted naphthyl groups. Biochem. Pharmacol. 56, 1243–1249 (1998).
Besson, M.J., Cheramy, A., Feltz, P. & Glowinski, J. Release of newly synthesized dopamine from dopamine-containing terminals in the striatum of the rat. Proc. Natl. Acad. Sci. USA 62, 741–748 (1969).
Scatton, B. & Lehmann, J. N-methyl-C-aspartate–type receptors mediate striatal 3H-acetylcholine release evoked by excitatory amino acids. Nature 297, 422–424 (1982).
Nguyen, M.L., Cox, G.D. & Parsons, S.M. Kinetic parameters for the vesicular acetylcholine transporter: two protons are exchanged for one acetylcholine. Biochemistry 37, 13400–13410 (1998).
Jentsch, T.J., Poet, M., Fuhrmann, J.C. & Zdebik, A.A. Physiological functions of CLC Cl− channels gleaned from human genetic disease and mouse models. Annu. Rev. Physiol. 67, 779–807 (2005).
Bankston, L.A. & Guidotti, G. Characterization of ATP transport into chromaffin granule ghosts. Synergy of ATP and serotonin accumulation in chromaffin granule ghosts. J. Biol. Chem. 271, 17132–17138 (1996).
Scherman, D. & Henry, J.P. Role of the proton electrochemical gradient in monoamine transport by bovine chromaffin granules. Biochim. Biophys. Acta 601, 664–677 (1980).
Hioki, H. et al. Chemically specific circuit composed of vesicular glutamate transporter 3– and preprotachykinin B–producing interneurons in the rat neocortex. Cereb. Cortex 14, 1266–1275 (2004).
Kawano, M. et al. Particular subpopulations of midbrain and hypothalamic dopamine neurons express vesicular glutamate transporter 2 in the rat brain. J. Comp. Neurol. 498, 581–592 (2006).
Yamaguchi, T., Sheen, W. & Morales, M. Glutamatergic neurons are present in the rat ventral tegmental area. Eur. J. Neurosci. 25, 106–118 (2007).
Gasnier, B. The loading of neurotransmitters into synaptic vesicles. Biochimie 82, 327–337 (2000).
Contant, C., Umbriaco, D., Garcia, S., Watkins, K.C. & Descarries, L. Ultrastructural characterization of the acetylcholine innervation in adult rat neostriatum. Neuroscience 71, 937–947 (1996).
Fujiyama, F. et al. Presynaptic localization of an AMPA-type glutamate receptor in corticostriatal and thalamostriatal axon terminals. Eur. J. Neurosci. 20, 3322–3330 (2004).
Bonsi, P. et al. Striatal metabotropic glutamate receptors as a target for pharmacotherapy in Parkinson's disease. Amino Acids 32, 189–195 (2007).
Consolo, S., Baldi, G., Giorgi, S. & Nannini, L. The cerebral cortex and parafascicular thalamic nucleus facilitate in vivo acetylcholine release in the rat striatum through distinct glutamate receptor subtypes. Eur. J. Neurosci. 8, 2702–2710 (1996).
Calabresi, P., Picconi, B., Parnetti, L. & Di Filippo, M. A convergent model for cognitive dysfunctions in Parkinson's disease: the critical dopamine-acetylcholine synaptic balance. Lancet Neurol. 5, 974–983 (2006).
Rogers, D.C. et al. Behavioral and functional analysis of mouse phenotype: SHIRPA, a proposed protocol for comprehensive phenotype assessment. Mamm. Genome 8, 711–713 (1997).
Crawley, J.N. Behavioral phenotyping of transgenic and knockout mice: experimental design and evaluation of general health, sensory functions, motor abilities and specific behavioral tests. Brain Res. 835, 18–26 (1999).
Fleming, S.M. et al. Early and progressive sensorimotor anomalies in mice overexpressing wild-type human α-synuclein. J. Neurosci. 24, 9434–9440 (2004).
Hamon, M. et al. Alterations of central serotonin and dopamine turnover in rats treated with ipsapirone and other 5-hydroxytryptamine1A agonists with potential anxiolytic properties. J. Pharmacol. Exp. Ther. 246, 745–752 (1988).
Kemel, M.L., Desban, M., Glowinski, J. & Gauchy, C. Distinct presynaptic control of dopamine release in striosomal and matrix areas of the cat caudate nucleus. Proc. Natl. Acad. Sci. USA 86, 9006–9010 (1989).
Paxinos, G. & Watson, C. The Mouse Brain in Stereotaxic Coordinates. (Academic Press, New York, 1997).
Blanchet, F. et al. Distinct modifications by neurokinin1 (SR140333) and neurokinin2 (SR48968) tachykinin receptor antagonists of the N-methyl-D-aspartate-evoked release of acetylcholine in striosomes and matrix of the rat striatum. Neuroscience 85, 1025–1036 (1998).
Huttner, W.B., Schiebler, W., Greengard, P. & De Camilli, P. Synapsin I (protein I), a nerve terminal-specific phosphoprotein. III. Its association with synaptic vesicles studied in a highly purified synaptic vesicle preparation. J. Cell Biol. 96, 1374–1388 (1983).
Herzog, E. et al. The existence of a second vesicular glutamate transporter specifies subpopulations of glutamatergic neurons. J. Neurosci. 21, RC181 (2001).
Ferguson, S.M. et al. Vesicular localization and activity-dependent trafficking of presynaptic choline transporters. J. Neurosci. 23, 9697–9709 (2003).
Kashani, A., Betancur, C., Giros, B., Hirsch, E. & El Mestikawy, S. Altered expression of vesicular glutamate transporters VGLUT1 and VGLUT2 in Parkinson disease. Neurobiol. Aging 28, 568–578 (2007).
Duplus, E. et al. Phosphorylation and transcriptional activity regulation of retinoid-related orphan receptor α1 by protein kinases C. J. Neurochem. published online, doi:10.1111/j.1471-4159.2007.05074.x (14 November 2007).
Acknowledgements
We thank S. Pérez, E. Etienne and L. Hillard for their excellent technical assistance with release experiments, confocal microscopy and animal care, respectively. This work was supported by grants from INSERM, Fédération pour la Recherche sur le Cerveau and Agence Nationale pour la Recherche. C.G. was supported by a fellowship from Fondation pour la Recherche Médicale and E.M.L. by France Alzheimer.
Author information
Authors and Affiliations
Corresponding author
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–4, Text and Methods (PDF 486 kb)
Rights and permissions
About this article
Cite this article
Gras, C., Amilhon, B., Lepicard, È. et al. The vesicular glutamate transporter VGLUT3 synergizes striatal acetylcholine tone. Nat Neurosci 11, 292–300 (2008). https://doi.org/10.1038/nn2052
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nn2052
This article is cited by
-
A conserved transcriptional fingerprint of multi-neurotransmitter neurons necessary for social behavior
BMC Genomics (2022)
-
Continuous cholinergic-dopaminergic updating in the nucleus accumbens underlies approaches to reward-predicting cues
Nature Communications (2022)
-
Opponent vesicular transporters regulate the strength of glutamatergic neurotransmission in a C. elegans sensory circuit
Nature Communications (2021)
-
Molecular, Structural, Functional, and Pharmacological Sites for Vesicular Glutamate Transporter Regulation
Molecular Neurobiology (2020)
-
Contribution of Vesicular Glutamate Transporters to Stress Response and Related Psychopathologies: Studies in VGluT3 Knockout Mice
Cellular and Molecular Neurobiology (2018)