An antiserum to gamma-aminobutyric acid (GABA) was used in a light and electron microscopic immunocytochemical study to determine the morphology and distribution of GABA-containing neurons in the rat visual cortex and to ascertain whether all classes of nonpyramidal neurons in this cortex are GABAergic. The visual cortex used for light microscopy was prepared in such a way that the antibody penetrated completely through tissue sections, and in these sections large numbers of GABA immunoreactive neurons were apparent. The labeled neurons could be identified as being either multipolar, bitufted, bipolar, or horizontal neurons. In layers II through VIa, GABA immunostained cells were distributed uniformly and accounted for approximately 15% of all neurons, but in layer I all neurons appeared to be immunostained. Electron microscopy of GABA immunostained visual cortex prepared to ensure good fine structural preservation confirmed the presence in layers II through VIa of numerous immunoreactive bipolar neurons, both small and large varieties, as well as multipolar and bitufted neurons. Additionally, electron microscopy reveals that astrocytes are frequently GABA immunoreactive. From a correlated light and electron microscopic evaluation of neurons in GABA immunostained visual cortex, it was possible to confirm which kinds of neurons are GABAergic and what proportion of the neuronal population they represent. Thus, from an analysis of some 950 neurons, it was found that pyramidal neurons were never immunoreactive and that except for 20% of the bipolar cell population, all examples of other types of nonpyramidal neurons encountered in this material were GABA immunoreactive.

Several lines of evidence suggest that the transient subplate zone of the embryonic mammalian telencephalon could influence cortical development through synaptic or trophic interactions with growing cortical afferents and migrating neurons. Since such interactions may involve neurotransmitters and their receptor molecules, we have examined the expression of GABA and subunits of the GABAA/benzodiazepine receptor complex in the occipital lobe of embryonic rhesus monkeys by immunochemistry and in situ hybridization. We found that during the second half of gestation, when the subplate zone reaches peak maturity in this species, many neurons can be immunolabeled with both GABA antisera and monoclonal antibodies against GABAA receptor subunits. The most robust labeling occurs at approximately embryonic day (E)125 (birth is at E165). Electron microscopic observations of receptor subunit-immunolabeled material confirmed that subunits of the GABAA receptor are localized in the subplate neurons and their dendritic processes. In many instances the reaction product is associated with the plasma membranes of labeled processes, some of which form symmetrical synapses with small unlabeled axon terminals. The results of in situ hybridization are in accord with the results of receptor subunit immunochemistry. From E80 to E141, hybridization signal for GABAA receptor subunit mRNA occurs in the subplate zone and increases steadily to peak levels between E125 and E141. The present results reveal that all the elements necessary for the formation of functional GABAergic synaptic circuitry are present in the subplate zone. Further, the ages showing the most pronounced receptor and transmitter expression in this primate coincide with the ingrowth of major cortical afferent systems. Taken together, these findings suggest that GABAergic local neuronal circuits in the subplate may be involved in the development of long tract connections stationed in this zone prior to their transfer to the overlying cortical plate.

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Light and electron microscopic immunohistochemistry was used to examine the time of emergence and distribution of GABA-containing cells in an association (prefrontal) and primary sensory (visual) region of the telencephalon at progressive fetal and postnatal stages of cortical maturation in the rhesus monkey. Thirty fetuses and six postnatal monkeys were examined beginning at embryonic day 41 (E41), the start of cortical neurogenesis, to birth (E165) and proceeding to maturity (greater than 5 years of age). The emergence and major developmental modifications in the distribution of immunoreactive neurons in both areas examined were nearly identical. GABA-immunolabeled neurons were present throughout the full thickness of the cerebral wall, including the cortical plate and the developmentally transient marginal, subplate, and ventricular zones, as early as E41. An important and surprising result was that a subset of bipolar migrating neurons in the subplate zone also contained GABA at these early stages. GABA-containing neurons in the ventricular and subventricular zones disappeared after E100, when neurogenesis is completed. In contrast, the number of immunoreactive multipolar and bipolar neurons within the subplate zone diminished between E100 and E131. By the first postnatal week, the distribution and density of GABA-containing neurons in the cortex appeared qualitatively similar to that observed in mature monkeys. The early appearance of GABA in cortical neurons and its expression by a population of migrating neurons suggest that a subset of cortical neurons may be committed to a transmitter phenotype independent of synaptic interactions and prior to attaining their adult positions in the maturing cerebral cortex.

When cat visual cortex (area 17) is reacted with an antibody to vasoactive intestinal polypeptide (VIP) a variety of neuronal types is labelled. Many of the labelled neurons are bipolar in form and are most common in layers II and III, although significant numbers of bipolar neurons are also encountered in layer V. Multipolar cells are also labelled. These are most frequent in layer IV and have a variety of shapes. In layer I, the labelled cells are of three varieties, i.e. horizontal bipolar cells, horizontal bitufted cells and multipolar neurons, while in layer VI the few VIP-positive neurons are horizontal bipolar cells. This suggests that all of the VIP-labelled neurons in cat area 17 are non-pyramidal in form, and this has been confirmed by electron microscopy. In these preparations, axon terminals are also labelled and under the light microscope it can be seen that these terminals occur both within the neuropil and around the cell bodies of some neurons, particularly neurons in layers II and III. Electron microscopy has shown that all of the labelled axon terminals form symmetric synapses and that those in the neuropil synapse with the shafts of smooth dendrites. These axodendritic synapses account for about 90% of the synapses formed by the labelled axon terminals. The remainder of the labelled axon terminals synapse with the cell bodies of pyramidal neurons. Parallels are drawn between these results and those previously obtained by examining those neuronal elements labelled with VIP antibodies in rat visual cortex.