Functional properties of strychnine-sensitive glycine receptors expressed in Xenopus oocytes injected with a single mRNA.
Mature rat spinal cord cDNA libraries constructed in lambda gt10 and lambda ZAPII were screened with an oligonucleotide probe (39 mer), and 4 clones that possess DNA-inserts encoding a glycine receptor subunit were obtained. The cloned cDNAs were used to reconstruct the nucleotide sequence of the full-length open reading frame consisting of 1350 base pairs (bp) as well as the 5'-(184 bp) and 3'-(591 bp) non-coding regions. Synthetic RNA transcribed in vitro from the glycine receptor cDNA induced Xenopus oocytes to synthesize functional glycine receptor that generated large Cl- currents. The electrophysiological properties of the wild-type receptor and some mutant receptors produced by site-directed mutagenesis were analyzed.
Akagi
H
H
Department of Neuroscience, Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Hirai
K
K
Hishinuma
F
F
D000109:Acetylcholine / Q000494:pharmacology; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D002712:Chlorides / Q000378:metabolism*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005998:Glycine / Q000378:metabolism*; D015640:Ion Channel Gating / Q000187:drug effects; D007473:Ion Channels / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000378:metabolism; D016366:Open Reading Frames; D010852:Picrotoxin / Q000494:pharmacology; D051381:Rats / Q000235:genetics; D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D013331:Strychnine / Q000494:pharmacology*; D014982:Xenopus laevis / Q000378:metabolism
D000109:Acetylcholine; D000596:Amino Acids; D002712:Chlorides; D004247:DNA; D005998:Glycine; D007473:Ion Channels; D010852:Picrotoxin; D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter; D011993:Recombinant Fusion Proteins; D013331:Strychnine