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Evidence for a third vesicular glutamate transporter isoform vglut3 cloning,functional characterization and distribution in mouse brain

Evidence for a third vesicular glutamate transporter isoform vglut3 cloning,functional characterization and distribution in mouse brain

Society for Neuroscience Abstract Viewer & Itinerary Planner : Abstract No 343 1

We have cloned and functionally identified a novel third isoform of the vesicular glutamate transporter (VGLUT3) from mouse brain. Studies using intracellular vesicles from transiently transfected PC12 cells indicate that uptake by VGLUT3 is highly selective for glutamate, H+-linked, and stimulated by Cl-ion. Northern Blot analysis revealed a low abundant VGLUT3 mRNA species of 4.8 kB expressed in brain. By Real Time PCR, low levels of VGLUT3 transcripts were detected throughout the adult mouse brain. Using in situ hybridization VGLUT3 mRNA was found at low cellular abundance in most brain areas with relatively high expression in distinct telencephalic, diencephalic and brain stem regions. Immunohistochemistry with affinity-purified polyclonal antisera revealed a low to moderate number of VGLUT3 immunopositive puncta and varicosities distributed throughout brain gray matter. Stained areas included basal forebrain nuclei, cerebral cortex, hippocampus, striatum and especially the diencephalon and brain stem. VGLUT3 synapses were clearly different from VGLUT1 or VGLUT2 stained synapses as shown by confocal double immunofluorescence. In addition, VGLUT3 expression was observed in the ependymal cells of the ventricles both by ISH and ICC pointing to a role of VGLUT3 at the brain liquor barrier. The molecular and functional characterization and the unique distribution of VGLUT3 strongly suppports the new concept of functional diversity of glutamatergic neurotransmission operated by at least three specific VGLUT isoforms.

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Accession: 034888016

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