期刊
EUROPEAN JOURNAL OF NEUROSCIENCE
卷 19, 期 9, 页码 2475-2487出版社
WILEY
DOI: 10.1111/j.0953-816X.2004.03349.x
关键词
clustering; cortex; development; hippocampus; GABA(A) receptors; synaptogenesis
We compared the expression and co-expression of alpha1, alpha2, alpha3, and alpha5-subunit protein clusters of the gamma-aminobutyric acid (GABA)(A) receptor in the neocortex and hippocampus of rat at postnatal days (PND) 5-10 and 30-40 in order to understand how inhibitory receptors reorganize during brain maturation. The size, intensity, density and pattern of co-localization of fluorescently tagged subunit clusters were determined in deconvolved digital images using a novel 2D cross-correlational analysis. The cross-correlation analysis allowed an unbiased identification of GABA(A) receptor subunit clusters based on staining intensity. Cluster size increased through development; only the alpha2 clusters in dentate gyrus (DG) decreased in size. alpha5-subunit cluster density either increased or decreased with maturation depending on the brain region. For the other subunits, the cluster density remained rather constant, with noted exceptions (increase in alpha2 clusters in cortical layer 5 but a decrease of alpha3 clusters in hilus). The co-localization of alpha1-subunit with the others was unique and not correlated to overall changes in subunit abundance between developmental epoques. So, although alpha2-subunit expression went up in the DG, the clusters became less co-localized with alpha1. In contrast, alpha5-subunit clusters became more co-localized with alpha1 as the alpha5-subunit expression declined in cortex and CA1. The co-localization of alpha3 with alpha1 also became greater in layer 6. In the adult brain not all clustering was associated with synapses, as many alpha-subunit clusters did not co-localize with synaptophysin. Overall, these data indicate that the regulation of GABA(A) receptor clustering is both synaptic and extrasynaptic, presumably reflecting complex cellular trafficking mechanisms.
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