Primary cultures of mouse spinal cord express the neonatal isoform of the inhibitory glycine receptor.
Expression of the inhibitory glycine receptor complex was investigated in primary cultures of fetal mouse spinal cord using sensitive immunomethods. In these cells, glycine receptor is predominantly of the neonatal isoform characterized by a low affinity for the antagonist strychnine. It contains a ligand binding subunit that differs from that of the adult receptor in antigenic epitopes and apparent molecular weight. Whereas in vivo the neonatal receptor isoform is completely replaced by the adult isoform within 3 weeks after birth, this exchange of subtypes is not seen in culture. The increased expression of the cytoplasmic glycine receptor-associated polypeptide of 93 kd occurring after birth is also seen under culture conditions. Purification of glycine receptor from cultures yielded polypeptides of 49 kd and 93 kd, suggesting that the membrane-spanning core of the neonatal receptor may be a homooligomer composed of 49 kd subunits. About half of the 49 kd subunit is cleaved by trypsinization of the cultures, indicating a predominant cell surface localization of the receptor. Pulse-labeling experiments revealed the 49 kd subunit to be a metabolically stable glycoprotein (half-life approximately 2 days). After its synthesis, a transition time of 30-45 min is required for acquisition of a strychnine binding conformation.
Hoch
W
W
Zentrum für Molekulare Biologie, Universität Heidelberg, Federal Republic of Germany.
Betz
H
H
Becker
C M
CM
D000818:Animals; D000831:Animals, Newborn; D046508:Culture Techniques; D005333:Fetus / Q000378:metabolism; D005998:Glycine / Q000378:metabolism; D007536:Isomerism; D008715:Methionine; D051379:Mice / Q000196:embryology; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D051381:Rats / Q000196:embryology; D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter / Q000302:isolation & purification / Q000378:metabolism*; D013116:Spinal Cord / Q000378:metabolism*; D013331:Strychnine / Q000378:metabolism
D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter; D008715:Methionine; D013331:Strychnine; D005998:Glycine