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Developmental regulation of expression of GABAA receptor alpha 1 and alpha 6 subunits in cultured rat cerebellar granule cells.

Abstract：

:We have studied the postnatal development of GABAA receptor alpha 1 and alpha 6 subunits expressed by primary cultures of cerebellar granule cells originating from 2-day-old (postnatal day 2, P2) and 10-day-old (P10) rat neonates. At these ages, the granule cells are at distinct stages of cerebellar development. In both cases, GABAA receptor alpha 1 and alpha 6 subunit-like immunoreactivities were detected, and displayed temporal expression profiles that were correlated with the maturity of the cerebella from which the cultured granule cells were derived. Using two different specificity anti-alpha 1 subunit-specific antibodies, immunoreactive species with M(r) 53,000 Da and 54,000 Da were detected by immunoblotting. The lower 53,000-Da band co-migrated with the alpha 1 subunit-like immunoreactivity detected in GABAA receptors purified from adult rat forebrain by benzodiazepine affinity chromatography. This 53,000-Da alpha 1 subunit-like immunoreactive species was detected at day 1 in vitro (1 DIV) in P10 cultures and 3-5 DIV in P2 cultures. The GABAA receptor alpha 6 subunit-like immunoreactivity (58,000 Da) was not detected until 5-7 DIV in P10 and 9-11 DIV in P2-derived cultures. The appearance of alpha 6 subunit-like immunoreactivity was paralleled by an up-regulation of alpha 1 subunit expression and a concomitant increase in diazepam-insensitive (DZ-IS) [3H]Ro 15-4513 binding activity, a pharmacological characteristic of alpha 6 and alpha 1 alpha 6-subunit-containing GABAA receptors (Pollard et al. J. Biol. Chem., 270, 21,285-21,290, (1995)). Antagonism of both non-NMDA and NMDA subtypes of ionotropic glutamate receptors did not significantly affect the developmental profile, the level of GABAA receptor alpha 6 subunit or the total DZ-IS or DZ-S [3H]Ro 15-4513 binding activities expressed by these neurons. These results provide further evidence that the expression of specific GABAA receptor subunit genes is subject to differential regulation. Furthermore, developmental expression of the GABAA receptor alpha 6 subunit gene by these neurons is either a preprogrammed event or is initiated by an environmental cue that is received early in granule cell development, and it is not a result of afferent activation of ionotropic glutamate receptors.

journal_name

Neuropharmacology

journal_title

Neuropharmacology

authors

Thompson CL,Pollard S,Stephenson FA

doi

10.1016/s0028-3908(96)00114-1

subject

Has Abstract

pub_date

1996-01-01 00:00:00

pages

1337-46

issue

9-10

eissn

0028-3908

issn

1873-7064

journal_volume

pub_type

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Developmental regulation of expression of GABAA receptor alpha 1 and alpha 6 subunits in cultured rat cerebellar granule cells.