GABA-synthesizing neurons were identified in the medulla of the rat by peroxidase-antiperoxidase (PAP) immunohistochemistry for glutamic acid decarboxylase (GAD). Using diaminobenzidine (DAB) either alone or intensified with silver, a relatively large number of GAD-immunoreactive neurons were evident within the reticular formation, raphe nuclei and vestibular nuclei. In all these areas, profuse GAD-immunoreactive varicosities appeared to contact the soma and dendrites of both non-GABA and GABA neurons. These observations suggest that GABA neurons may act as interneurons or local projection neurons within the medulla and accordingly exert a potent inhibitory and/or disinhibitory control on bulbar projection neurons. Within the ventral reticular formation (pars alpha and ventralis of the gigantocellular reticular field) and raphe magnus, large numbers of prominent GAD-immunoreactive neurons resembled in size and morphology and overlapped in distribution the serotonin-immunoreactive neurons of the same regions. However, by sequential double immunostaining utilizing DAB as a chromogen for serotonin (5-HT) and benzidine dihydrochloride (BDHC) for GAD, it was found that GAD-containing neurons were distinct from 5-HT-containing neurons. Following injections of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) into the upper cervical spinal cord and combined processing for WGA-HRP (using tetramethylbenzidine [TMB] with cobalt) and immunohistochemistry (with DAB), a contingent of spinally projecting neurons were found to contain GAD. The GAD-immunoreactive reticulo- and raphe-spinal neurons were most frequent within the pars alpha and ventralis of the gigantocellular reticular fields and the raphe magnus, where they were approximately equal in number to the coexistent, but distinct 5-HT spinally projecting neurons. GABA neurons of the medulla may thus contribute directly to the bulbar inhibitory influence upon spinal sensory and motor systems.

The brainstem mechanisms for the generation of paradoxical sleep are under considerable debate. Previous experiments in cats have demonstrated that injections of the cholinergic agonist carbachol into the oral pontine tegmentum elicit paradoxical sleep behaviour and its polygraphic correlates. The different results on the pontine structures that mediate this effect do not agree. We report here that limited microinjections of a carbachol solution into the ventral part of the oral pontine reticular nucleus in the cat induce, with a short latency, a dramatic, long-lasting increase in paradoxical sleep. Moreover, neuronal tracing experiments show that this pontine site is connected with brain structures responsible for the different bioelectric events of paradoxical sleep. These two facts suggest that the ventral part of the oral pontine reticular nucleus is a nodal link in the neuronal network underlying paradoxical sleep mechanisms.

Double bouquet cells (DBCs) are neocortical gamma-aminobutyric acid (GABA)ergic interneurons characterized by the vertical bundling of its axon, which are generally termed "bundles" or "horse-tails." Using immunocytochemistry for the calcium binding protein calbindin, we have analyzed the morphology, density, and distribution of DBC horse-tails in different cortical areas of the human cortex (Brodmann's areas 10, 4, 3b, 22, 18, and 17). Although DBC horse-tails were very numerous and regularly distributed in all cortical areas, variations were observed both in terms of morphology and density. We distinguished two major classes of DBC horse-tails: the thicker complex type (type I) that had more axon collaterals; and the simple type (type II). The density of DBC horse-tails was significantly higher in areas 17, 18, 22, and 4 than in areas 3b and 10. Moreover, the proportion of type I and type II DBC horse-tails varied in the cortical areas studied. We also examined the distribution of DBC horse-tails in frontal, parietal, and occipital areas of different mammalian species. We found DBCs to be present in carnivores but not in rodents, lagomorphs, or artiodactyls. In carnivores, relatively few DBC horse-tails can be identified and they were generally found in the occipital cortex. Therefore, there is significant variability in the morphology and distribution of DBC horse-tails in different species and cortical areas. We conclude that, although these interneurons may be an important element in the organization of cortical microcolumns in primates, this is not the case in other mammalian species.

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