GABA transporter mRNA: in vitro expression and quantitation in neonatal rat and postmortem human brain.
A previously isolated rat cDNA clone encoding the membrane transporter for the neurotransmitter gamma-aminobutyric acid was expressed in transfected COS cells. The resultant transporter protein was characterized kinetically and pharmacologically. The apparent Kt (6.1 microM) and the pharmacological profile of a neuronal-type transporter observed in these mammalian cells were consistent with previous data obtained in Xenopus laevis oocytes. Post-natal levels of gamma-aminobutyric acid transporter mRNA in rat cerebellum, cerebral cortex and striatum (as measured by nuclease protection assay) transiently exceeded levels present in the adult brain. Human gamma-aminobutyric acid transporter mRNA also was measured by nuclease protection assay using as probe a human transporter cDNA homolog obtained by polymerase chain reaction. These studies suggest that quantitation of rat and human gamma-aminobutyric acid transporter mRNAs may provide a useful index of transporter gene expression.
Xia
Y
Y
Department of Psychiatry, Wayne State University School of Medicine, Detroit, Michigan.
Poosch
M S
MS
Whitty
C J
CJ
Kapatos
G
G
Bannon
M J
MJ
D000328:Adult; D000375:Aging / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D002102:Cadaver; D002352:Carrier Proteins / Q000235:genetics*; D002478:Cells, Cultured; D005260:Female; D050485:GABA Plasma Membrane Transport Proteins; D006801:Humans; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000235:genetics*; D026901:Membrane Transport Proteins; D008875:Middle Aged; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D027361:Organic Anion Transporters; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012260:Ribonucleases; D014018:Tissue Distribution; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D002352:Carrier Proteins; D050485:GABA Plasma Membrane Transport Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D015335:Molecular Probes; D027361:Organic Anion Transporters; D012333:RNA, Messenger; D005680:gamma-Aminobutyric Acid; D012260:Ribonucleases