Developmental changes of agonist affinity at GABABR1 receptor variants in rat brain.
Recently, two N-terminal splice variants of the metabotropic receptor for GABA (gamma-amino-butyric acid) were cloned. Here, we describe an antiserum that recognizes the two receptor variants. We demonstrate that these proteins are identical with GABAB receptors that are photoaffinity labeled with [125I]CGP71872 in rat brain. The C-terminal epitopes recognized by the antiserum are conserved in several vertebrate species but not in chicken. No hints for the existence of additional closely related receptor subtypes or variants are found in double-labeling experiments with antibody and photoaffinity ligand. Western blot analysis reveals widespread expression of the GABABR1 receptor proteins in rat brain with the highest level of expression at early postnatal stages. The binding affinity of the GABAB receptor agonist L-baclofen at native R1a and R1b variants is similar. In early postnatal development the affinity at R1a and R1b is 10-fold lower than in adult brain and gradually increases with aging.
TA Nervous System, Novartis Pharma AG, Basel, CH-4002, Switzerland.
B
B
Malitschek
D
D
Rüegg
J
J
Heid
K
K
Kaupmann
H
H
Bittiger
W
W
Fröstl
B
B
Bettler
R
R
Kuhn
D000818:Animals; D001386:Azides / Q000494:pharmacology*; D001418:Baclofen / Q000494:pharmacology; D001923:Brain Chemistry / Q000502:physiology*; D002460:Cell Line; D002645:Chickens; D018755:GABA Agonists / Q000494:pharmacology; D018756:GABA Antagonists / Q000494:pharmacology*; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007457:Iodine Radioisotopes; D007536:Isomerism; D007668:Kidney / Q000166:cytology; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D011233:Precipitin Tests; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D017434:Protein Structure, Tertiary; D011869:Radioligand Assay; D051381:Rats; D018080:Receptors, GABA-B / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology; D014162:Transfection
D001386:Azides; C104980:CGP 71872; D018755:GABA Agonists; D018756:GABA Antagonists; D007457:Iodine Radioisotopes; D009943:Organophosphorus Compounds; D018080:Receptors, GABA-B; D001418:Baclofen