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Glutamate vesicular transporter bNPI and putative excitatory amino acid vesicular transporter dNPI show differential distribution in neurons projecting to cerebellar cortex in rat



Glutamate vesicular transporter bNPI and putative excitatory amino acid vesicular transporter dNPI show differential distribution in neurons projecting to cerebellar cortex in rat



Society for Neuroscience Abstracts 27(1): 1506



The bNPI gene encodes a vesicular glutamate transporter and is expressed in brainstem. The dNPI gene is 81% identical to bNPI and encodes a putative EAA vesicular transporter. The present study seeks to determine whether neurons projecting to cerebellar cortex contain bNPI or dNPI mRNA. cDNAs for bNPI and dNPI (apprx1000 bp) were obtained by RT-PCR using specific primers for bNPI or dNPI sequence with rat brain polyA+ RNA as template. The PCR product specific for each gene was subcloned into pCRII-Topo(R) plasmid vector used as template for synthesizing digoxigenin-tagged riboprobes for detection of bNPI or dNPI mRNA by in situ hybridization. Fluoro-Gold (FG) was injected in cerebellar cortex of 4 rats. The vast majority of FG retrogradely labeled neurons in brainstem and spinal cord contained bNPI mRNA except the inferior olivary nucleus where FG was exclusively colocalized with dNPI mRNA. bNPI mRNA and FG colocalized in neurons in the following nuclei: linearis, prepositus, lateral reticular, spinal trigeminal (interpolar), external cuneate, pontine, reticular tegmental pontine and Clarke's column in spinal cord. All these nuclei are sources of mossy fibers known to use an EAA transmitter, probably glutamate. The inferior olive is the source of climbing fibers known to use an EAA. The current data support the idea that dNPI mRNA is a marker for a specific class of EAA-releasing neurons and that climbing fibers use a different EAA vesicular transporter and possibly different transmitter than mossy fibers.

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