Seven monkey-specific GABAA receptor subunit cDNAs were isolated and cloned; radioactive cDNA and cRNA probes derived from them were used for Northern blot analysis and in situ hybridization histochemistry of the primary visual cortex (area 17), with comparative observations on other cortical areas. cDNAs corresponding to alpha 1, alpha 2, alpha 4, alpha 5, beta 1, beta 2, and gamma 2 GABAA receptor subunits were isolated and had sequences unique to the monkey but recognized mRNAs of distinct molecular weights consistent with those reported in other species. mRNAs for the alpha 1, beta 2, and gamma 2 subunits were expressed at much higher levels in area 17 than in motor, somatosensory, or temporal association cortex, possibly reflecting the greater density of GABA cells and synapses in area 17. In areas 17 and 18, each of the seven subunit mRNAs showed individually distinct patterns of laminar distribution. alpha 1, beta 2, and gamma 2 subunit mRNAs, which are thought to form the basis of receptors with the full range of classical GABAA receptor properties in the adult, were particularly enriched in layers II-III, IVC, and VI of area 17, following patterns of receptor distribution previously demonstrated by radioligand binding and immunocytochemistry. alpha 2, alpha 4, alpha 5, and beta 1 transcripts had quite different localization patterns that did not match the antoradiographic or immunocytochemical receptor localization patterns. alpha 2 and alpha 5 subunit mRNAs, which are thought to be the subunits mainly expressed in development, were enriched in layer VI and the underlying white matter, possibly reflecting the involvement of receptors formed from alpha 2 and alpha 5 polypeptides in trophic interactions in the cortical subplate zone during development of the cerebral cortex. Following 8–21 d periods of monocular deprivation induced by intravitreal injection of TTX, levels of alpha 1, beta 2, and gamma 2 subunit mRNAs were substantially reduced in deprived ocular dominance columns of layer IVC in area 17. The effect was greatest for the alpha 1 subunit; for both alpha 1 and gamma 2 subunit mRNAs, it extended into deprived rows of cytochrome oxidase-identified periodicities in other layers. Apart from the alpha 5 subunit mRNA, which showed reduced levels in layer VI, the other subunit mRNAs were unaffected by monocular deprivation. These results demonstrate the heterogeneity of GABAA receptor subunit expression in a complex, multilaminar cortical area. They suggest that receptors with different functional properties may be assembled from different combinations of subunit polypeptides in different layers and show that subunit expression is differentially regulated under activity-dependent conditions.