Differences between GABA Receptor Binding to Membranes from Cerebellum during Postnatal Development and from Cultured Cerebellar Granule Cells

Abstract

GABA receptor binding sites were determined in membranes from cerebella of 7-, 15- and 60-day-old rats and cerebellar granule cells derived from 7-day-old rats and cultured for 8 days using [³H]GABA as the ligand and nonradioactive GABA to measure non-specific binding. Membranes from cerebellum at all postnatal stages exhibited at least two binding sites for GABA with binding constants around 7–9 and 150–750 nM, respectively. The total number of binding sites expressed on the basis of total protein increased 10 times between 7 and 60 days of age and 2 times between 15 and 60 days. The latter increase could be quantitatively accounted for by an increase in the number of low affinity sites. In contrast to this, membranes from cultured granule cells only exhibited the high affinity binding site (K_D 7.1±0.5 nM) and the number of binding sites was comparable to that of cerebellar membranes from rats of the corresponding age (15 days old). GABA analogues such as muscimol, piperidine-4-sulphonic acid, homo-β-proline, THIP, and isoguvacine were potent displacers of GABA binding to cultured granule cells, whereas nipecotic acid and guvacine had no effect.

Cell type‐specific regulation of major histocompatibility complex (MHC) Class I gene expression in astrocytes, oligodendrocytes, and neurons

“…Neuronal cultures were produced from either embryonic day-15 cerebral hemispheres or from 8-day postnatal mouse cerebellum of BIO.A mice as previously described (Meier and Schousboe, 1982;Yu et al, 1984). Neurons were analyzed at 8 days of culture.…”

Pharmacological in vitro characterization of the arecoline bioisostere, Lu 25-109-T, a muscarinic compound with M1-agonistic and M2/M3-antagonistic properties

“…in 24 multi-well poly-L-lysine-coated tissue culture dishes as described by Meier and Schousboe [1982] except that the medium was not changed after addition of 20 µM of the mitotic inhibitor, cytosine-arabinoside. The cultures were used for experiments after 5-9 days in culture.…”

Experimental studies of the influence of vigabatrin on the GABA system.

“…Interneurons from cerebral cortex of 15 day old mouse embryos were cultured as previously described (Dichter, 1978;Larsson et al, 1981;Yu et al, 1984) with the modifications described by Meier & Schousboe (1982). After 48 h in culture the cells were exposed to cytosine arabinoside, which leads to the disappearance of astroblasts (Dichter, 1978;Yu et al, 1984).…”