The RNA of the glutamate transporter EAAT2 is variably spliced in amyotrophic lateral sclerosis and normal individuals.
Impaired re-uptake of synaptic glutamate, and a reduced expression of the glutamate transporter EAAT2 have been found in the motor cortex of patients with amyotrophic lateral sclerosis (ALS). Two splice forms of the EAAT2 RNA resulting from retention of intronic sequences (EAAT2/Int) and deletion of one protein coding exon (EAAT2/C1) have been reported to account for the EAAT2 protein loss in ALS. In this study we investigated the presence of two known (EAAT2/C1; EAAT2/Int) and three novel (EAAT2/C2-4) EAAT2 RNA in motor cortex of 17 ALS cases and 11 controls. Reverse transcription and PCR were carried out to amplify the complementary DNA of the complete and variably spliced EAAT2 transcripts. Nested PCR was followed to generate amplicons specific for EAAT2/C1-4 and EAAT2/Int. EAAT2/Int was detected in 59% of ALS specimens as compared to 36% of controls showing a trend but no statistical significance of a more frequent expression in ALS (Type I error 24.6%). EAAT2/C1-4 were found to be equally expressed in ALS patients and controls. Our results indicate that the involvement of EAAT2 transcripts in ALS is unlikely to be primary, and more complex than previously recognized. Alterations of quantitative expression of distinct EAAT2 splice forms in ALS cannot be excluded from this study and remain to be investigated.
Meyer
T
T
Department of Neurology, University of Ulm, Albert-Einstein-Allee 11, D-89081, Ulm, Federal Republic of Germany. thomas.meyer@medizin.uni-ulm.de
Fromm
A
A
Münch
C
C
Schwalenstöcker
B
B
Fray
A E
AE
Ince
P G
PG
Stamm
S
S
Grön
G
G
Ludolph
A C
AC
Shaw
P J
PJ
D000368:Aged; D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics* / Q000378:metabolism; D002102:Cadaver; D027342:Excitatory Amino Acid Transporter 2; D014644:Genetic Variation; D006801:Humans; D009044:Motor Cortex / Q000378:metabolism; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D012326:RNA Splicing; D017981:Receptors, Neurotransmitter / Q000235:genetics*; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction
D027342:Excitatory Amino Acid Transporter 2; D017981:Receptors, Neurotransmitter; D012313:RNA