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Journal of Neuroscience, Vol 15, 1835-1853, Copyright © 1995 by Society for Neuroscience
Differential expression of two glial glutamate transporters in the rat brain: quantitative and immunocytochemical observations
KP Lehre, LM Levy, OP Ottersen, J Storm-Mathisen and NC Danbolt
Anatomical Institute, University of Oslo, Blindern, Norway.
Glutamate, the major excitatory neurotransmitter in brain, is almost
exclusively intracellular due to the action of the glutamate transporters
in the plasma membranes. To study the localization and properties of these
proteins, we have raised antibodies specifically recognizing parts of the
sequences of two cloned rat glutamate transporters, GLT-1 (Pines et al.,
1992) and GLAST (Storck et al., 1992). On immunoblots the antibodies
against GLT-1 label a broad heterogeneous band with maximum density at
around 73 kDa, while the antibody against GLAST labels a similarly broad
band at around 66 kDa in the cerebellum and a few kilodaltons lower in
other brain regions. GLT-1 is expressed at the highest concentrations in
the hippocampus, lateral septum, cerebral cortex, and striatum, while GLAST
is preferentially expressed in the molecular layer of the cerebellum.
However, both transporters are present throughout the brain, and have
roughly parallel distributions in the cerebral hemispheres and brainstem.
Preembedding light and electron microscopical immunocytochemistry shows
that both GLT-1 and GLAST are restricted to astrocytes, which appear to
express both proteins concomitantly, but in different proportions in
different parts of the brain. Nerve terminal labeling was not observed.
Both the amino and carboxyl terminals of GLT- 1 and GLAST are located
intracellularly, indicating an even number of transmembrane segments.
Antibodies against a synthetic peptide corresponding to amino acid residues
2-11 of the proposed sequence of GLT-1 recognize the native rat brain GLT-1
protein, confirming that the translation initiation site is at the first
ATG.
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