A splice variant of glutamate transporter GLT1/EAAT2 expressed in neurons: cloning and localization in rat nervous system.
Rapid uptake of synaptically released glutamate via the high affinity glutamate transporter 1 (GLT1; EAAT2) is important for limiting transmitter signalling and prevents a harmful receptor overstimulation. So far, in the adult brain GLT1 protein has only been detected in astrocytes. Here, we describe the cDNA cloning of a variant of GLT1 from rat brain which is generated by alternative splicing at the 3'-end of the GLT1 cDNA. Reverse transcription-polymerase chain reaction revealed that the GLT1 variant message was present not only in brain, but also in peripheral organs. Northern blot analysis showed that in brain the mRNA of GLT1 (approximately 11 kb) is predominant while in the retina the mRNA of GLT1 variant (approximately 12.5 kb) prevails. In situ hybridization using cRNA and oligonucleotide probes, and immunocytochemistry using an antibody against a synthetic GLT1v peptide were applied in order to identify the cell types expressing GLT1 variant in the adult rat nervous system. GLT1 variant is preferentially expressed in neurons of the CNS and PNS, but is also detected in glial cells (oligodendrocytes, ependymal cells, epithelial cells of the plexus choroideus, satellite cells, and Schwann cells). In contrast to GLT1, GLT1 variant was only occasionally detected in astrocytes. Immunolabelling revealed a preferentially cytoplasmic (frequently granular) staining of neurons and glial cells, suggesting a localization of GLT1 variant protein in vesicle membranes. The studies provide evidence that the cellular expression of the GLT1 variant in the CNS is almost complementary to that of GLT1 and that the GLT1 variant does not seem to be restricted to the CNS.
Schmitt
A
A
Institute of Anatomy and Cell Biology, University of Würzberg, Koellikerstrasse 6, D-97070 Würzberg, Germany.
Asan
E
E
Lesch
K-P
KP
Kugler
Peter
P
D017398:Alternative Splicing / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002831:Choroid Plexus / Q000166:cytology / Q000378:metabolism; D003001:Cloning, Molecular; D027342:Excitatory Amino Acid Transporter 2 / Q000235:genetics* / Q000378:metabolism*; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism*; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D009197:Myenteric Plexus / Q000166:cytology / Q000378:metabolism; D009420:Nervous System / Q000166:cytology / Q000378:metabolism*; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000166:cytology / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012583:Schwann Cells / Q000166:cytology / Q000378:metabolism; D012584:Sciatic Nerve / Q000166:cytology / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism
D027342:Excitatory Amino Acid Transporter 2; D020033:Protein Isoforms; D012333:RNA, Messenger; D018698:Glutamic Acid