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Journal of Neuroscience, Vol 14, 5302-5324, Copyright © 1994 by Society for Neuroscience
Switch in the expression of rat GABAA-receptor subtypes during postnatal development: an immunohistochemical study
JM Fritschy, J Paysan, A Enna and H Mohler
University of Zurich, Institute of Pharmacology, Switzerland.
The involvement of GABA in neuronal differentiation and maturation precedes
its role as inhibitory neurotransmitter in the brain. It was therefore
investigated whether GABAA receptors mediating the actions of GABA in
neonatal and adult brain can be distinguished by their molecular structure
and cellular location. Immunohistochemistry with subunit-specific
antibodies was employed to analyze changes in the distribution of
GABAA-receptor subunits during postnatal development. In particular,
subunit association patterns, as evidenced by colocalization of subunits
within individual neurons, were analyzed by confocal laser microscopy. The
subunits analyzed include the alpha 1- and alpha 2-subunits, which are
associated with pharmacologically distinct GABAA-receptor subtypes, and the
beta 2,3-subunits, which are a major constituent of GABAA receptors in both
immature and adult rat brain. Each of these subunits exhibited
age-dependent changes in their distribution, indicative of a differential
maturation process. The alpha1-subunit immunoreactivity (-IR) was low at
birth, restricted to a few areas, and increased dramatically during the
first postnatal weeks. By contrast, the alpha 2-subunit-IR displayed a
widespread distribution throughout the brain at birth, and disappeared from
numerous areas soon after the appearance of the alpha 1-subunit.
Double-immunofluorescence staining demonstrated the coexistence of both
subunits in many individual neurons during a short time window, indicating
that receptors containing the alpha 1-subunit gradually replace receptors
containing the alpha 2-subunit in these cells. Staining for the beta
2,3-subunits was prominent and ubiquitous at every developmental age,
indicating that these subunits are present in both neonatal and adult GABAA
receptors. Indeed, double-immunofluorescence staining revealed an extensive
colocalization of the alpha 2- and beta 2,3-subunits in neurons from
neonatal rats, whereas the beta 2,3-subunits were associated with the alpha
1-subunit at later stages. Thus, the onset of alpha 1-subunit staining in
maturing brain is indicative for the expression of a new, prevalent
receptor subtype, presumably involved in synaptic inhibition. These
findings demonstrate a switch in the subunit composition of GABAA receptors
during postnatal development, suggesting the existence of molecularly
distinct immature and adult forms of GABAA receptors in rat CNS.
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