Bacterial lipopolysaccharide-induced intestinal microvascular lesions leading to acute diarrhea.

Subcutaneous challenge of mice with lipopolysaccharide (LPS) from gram negative bacteria, produced an intestinal microvascular lesion causing fluid exudation into the lumen of the intestine and diarrhea. The microvascular lesion was characterized by endothelial cell damage and microthrombi in the venules and capillaries of the intestinal lamina propria. Marker organisms, given orally to challenged mice, grew in the exuded fluid and could invade the mucosa. Intravenous transfer of postchallenge plasma produced the lesion in normal mice and absorption of such plasma by Sepharose coupled to LPS-antibody abolished this effect. Instillation of large quantities of LPS into the lumen of the intestine produced scattered microvascular lesions, although none of these animals developed diarrhea. Since a similar microvascular lesion has been described in the rectal mucosal lamina propria of adults with acute diarrhea, it is suggested that LPS-induced vascular damage may be a novel mechanism in the pathogenesis of acute diarrhea.

The 'marginal division': a new subdivision in the neostriatum of the rat.

Using a combination of anterograde and retrograde (Phaseolus vulgaris leucoagglutinin; PHA-L and wheat germ agglutinin conjugated horseradish peroxidase; WGA-HRP) tract-tracing methods and histochemical techniques, a new subdivision of the neostriatum, the marginal division, has been found in the rat brain. The marginal division is approximately 120 microns wide and is located at the caudal extent of the neostriatum and surrounds the rostral edge of the globus pallidus. The neuronal somata of the marginal division are mostly fusiform in shape, with their long axes running parallel to the border between the striatum and the globus pallidus. Histochemically, the marginal division is lighter in AChE staining, is more densely filled with Met-enkephalin-immunoreactive terminals, and has fewer choline acetyltransferase (ChAT)-immunoreactive neurons than does the rest of the neostriatum. Injections of PHA-L or WGA-HRP demonstrated that the projections of the marginal division differ from those of the main body of the striatum. The striatopallidal projection from the marginal division terminates in the caudal-most part of the globus pallidus which is rich in cholinergic neurons. In contrast, the projection from the main region of the neostriatum terminates in two bands in the globus pallidus, both of which are rostral to the area of termination of the fibres from the marginal division. The striatonigral fibres from the marginal division terminate in the caudal part of the substantia nigra pars reticulata whereas the rest of neostriatum projects to a more rostral region. Based on its cellular morphology, immunohistochemistry and projection pattern, we conclude that the marginal division of the striatum is a distinct subdivision of the neostriatum.

Relationship of the axonal and dendritic geometry of spiny projection neurons to the compartmental organization of the neostriatum.

Intracellular injection of HRP combined with immunocytochemistry for [Leu]enkephalin was used to demonstrate striatal spiny neuron dendritic and local axonal arborizations in the same section as enkephalin-rich patches (striosomes). Cobalt intensification of the first DAB reaction prior to the immunoperoxidase steps resulted in good contrast between the black reaction product in the intracellularly labeled cells and the brown staining for [Leu]enkephalin. Serial reconstructions of the labeled cells and nearby boundaries between the enkephalin-rich striosomes and enkephalin-poor matrix allowed the relationship between the arborizations of the labeled cells and these boundaries to be established. It was also possible to examine the relationship to compartmental boundaries of a second neuronal class consisting of large, pallidallike neurons whose somatodendritic morphology was outlined by immunoperoxidase-labeled terminals. We found that spiny projection neurons in both compartments have dendritic arbors and local axonal collaterals that are confined by compartmental boundaries. The termination or recurvature of dendrites at such boundaries suggests that the cellular basis of striatal compartmental organization is provided by this class of striatal neuron. On the other hand, large pallidumlike striatal neurons were found to have dendrites that extend across compartmental boundaries. These results support previous reports that striatal spiny projection neurons preserve the compartmental segregation of parallel striatal input-output systems, whereas other classes of striatal neurons may serve to provide limited integration between compartments.

Enkephalinergic-cholinergic interaction in the rat globus pallidus: a pre-embedding double-labeling immunocytochemistry study.

The synaptic relationships between leucine-enkephalin containing axon terminals and cholinergic neurons in the rat globus pallidus were studied at both light and electron microscopic levels using a high resolution pre-embedding double-labeling immunocytochemical method. Results indicated that leucine-enkephalin terminals very rarely form monosynaptic connections with cholinergic neurons in the rat globus pallidus, suggesting that enkephalinergic neostriatal efferents probably have little monosynaptic influences on the activities of pallidal cholinergic neurons.

Terminations of individual optic tract fibers in the lateral geniculate nuclei of Galago crassicaudatus and Tupaia belangeri.

The morphology and laminar distribution of individual optic fibers projecting to the lateral geniculate nucleus (GL) of Galago and Tupaia were studied following iontophoretic injections of horseradish peroxidase (HRP) into the optic tract. In Galago the GL is composed of three functionally matched pairs of layers, each characterized by cells of a given size, one large, one medium-sized, and one small. The results show that there is a close correspondence between the size of the afferent fibers and the size of the neurons in the target layer: large axons project to the magnocellular layers, medium-sized axons project to the parvicellular layers, and small fibers project to the intercalated layers. In Tupaia the GL is composed of two functionally matched pairs and two unmatched layers. Optic fibers that project to the medial matched pair (1 and 2) are only slightly larger than those that project to the lateral matched pair (4 and 5), but both are larger than those that project to the unmatched layers (3 and 6). In both species terminal arbors and the distribution of terminal boutons within layers corresponded closely with the organization of dendritic processes of cells in the target layer. This correspondence was particularly evident in the parvicellular layers in Galago and in layer 6 in Tupaia: parvicellular terminal arbors, like the dendrites of parvicellular cells, are organized in narrow columns oriented along lines of projection, whereas layer 6 terminal arbors, like the dendrites of layer 6 cells, are oriented in elongated strips perpendicular to lines of projection. In both species there was evidence for sublaminar terminations in some layers. These were restricted to the parvicellular layers in Galago and layers 4 and 5 in Tupaia. With the exception of a small number of fine fibers in the intercalated layers in Galago, optic fibers in both species terminated in one and only one layer in a set. The significance of this result depends on the relation between ganglion cell classes and what is being segregated in different GL layers. Lateral geniculate lamination varies even in closely related species and has evolved independently in such distantly related lines as carnivores and primates. It is not surprising, therefore, that what is being segregated varies from species to species.

The glutamate decarboxylase-, leucine enkephalin-, methionine enkephalin- and substance P-immunoreactive neurons in the neostriatum of the rat and cat: evidence for partial population overlap.

We have examined the populations of neurons in the neostriatum of both rat and cat that are immunoreactive for glutamate decarboxylase, [Leu]enkephalin, [Met]enkephalin and substance P. Neurons that were immunoreactive for glutamate decarboxylase made up 47% of the neurons in our samples from the rat and ranged from 39 to 49% of the neurons in the cat. Those immunoreactive for [Leu]enkephalin made up 44-49% of the neurons in rat neostriatum, and 38-47% in the cat, and those immunoreactive for [Met]enkephalin made up 36-41% of the neurons in rat and 43-49% of the neurons in the cat. Substance P-immunoreactive neurons made up 30-38% of neurons in rat and 32-39% in cat. Most substance P neurons (particularly the most darkly staining ones) were, however, clustered such that they were most numerous in the patch compartment of neostriatum; within the patches the substance P neurons comprised 59% of neurons in the rat and 55% in cat, but in the matrix substance P neurons comprised only 32% of neurons in the rat and 25% in the cat. Samples taken from sections processed for two-color double labeling immunocytochemistry revealed that 12% of neurons label for both glutamate decarboxylase and [Leu]enkephalin, 12% for both glutamate decarboxylase and [Met]enkephalin, 11-12% for both glutamate decarboxylase and substance P, and 17% for both [Met]enkephalin and substance P. These results provide evidence for chemical heterogeneity within the medium-sized neostriatal neurons, and provide the first evidence for coexistence of glutamate decarboxylase and substance P within a single neuron, and the first evidence for the coexistence for substance P and [Met]enkephalin within single neurons of the central nervous system.

Substance P-immunoreactive neurons in the neocortex of the rat: a subset of the glutamic acid decarboxylase-immunoreactive neurons.

Substance P-immunoreactive (SP-IR) neurons are found throughout the layers of the rat neocortex. Within the somatosensory (SI) cortex these cells make up about 2% of all neurons and within area 17 about 3%. Colocalization studies reveal that the SP-IR neurons are a subset of the population immunoreactive for glutamic acid decarboxylase.

The distribution of glutamic acid decarboxylase immunoreactivity in the diencephalon of the opossum and rabbit.

We have examined the distribution of neurons and terminals immunoreactive for glutamic acid decarboxylase (GAD) in the thalamus and adjacent structures of the opossum (Didelphis virginiana) and the rabbit and have compared this distribution with the distributions we described previously for the cat and bushbaby (Galago senegalensis). The significance of these experiments depends, first, on the fact that GAD is the synthetic enzyme for GABA, and therefore that GAD immunoreactivity is a marker for GABAergic inhibitory neurons, and second, on previous findings that suggest that GABAergic neurons in the dorsal thalamus are local circuit neurons. In both cat and Galago, GAD-immunoreactive neurons are distributed essentially throughout the entire thalamus. In the opossum, GAD neurons are chiefly confined to the dorsal lateral geniculate nucleus and the lateral extremity of the lateral posterior nucleus. The distribution of GAD neurons in the rabbit is intermediate between that found in the opossum on the one hand and cat and Galago on the other. Like opossum, about 25% of the neurons in the lateral geniculate nucleus of rabbit are GAD immunoreactive. Unlike opossum, however, as many as 18% of the cells in the ventral posterior nucleus of the rabbit are GAD immunoreactive, and scattered cells are also labeled in other thalamic areas, such as the medial geniculate and the lateral group. Aside from the findings in the dorsal thalamus, the chief observation is that GAD-immunoreactive neurons and/or terminals densely fill all principal targets of the optic tract, including the ventral lateral geniculate nucleus; the superficial gray layer of the superior colliculus; the anterior, posterior, and olivary pretectal nuclei; the nucleus of the optic tract; and the medial and lateral terminal nuclei of the accessory optic tract. These results support the idea first put forward by Cajal that local circuit neurons increase in number during the course of the evolution of complex mammalian brains. If we can assume that the conservative opossum retains characteristics reflecting an early stage of mammalian evolution, the results suggest that thalamic local circuit neurons arose first in the visual system and only later in evolution spread throughout the thalamus.

Glutamic acid decarboxylase-immunoreactive neurons and terminals in the lateral geniculate nucleus of the cat.

We have examined the distribution of neurons and terminals that are immunoreactive for glutamic acid decarboxylase (GAD), the synthesizing enzyme for the inhibitory neurotransmitter gamma-aminobutyric acid within the lateral geniculate nucleus of the cat. We estimate that GAD-positive neurons constitute approximately one-fourth of the neurons in all layers of the lateral geniculate nucleus and in the medial interlaminar nucleus (MIN). In addition, almost all of the neurons within the perigeniculate nucleus are GAD-positive. The mean size of GAD-positive cell bodies is significantly smaller than the mean size of unlabeled neurons in all subdivisions of the lateral geniculate nucleus. GAD-positive neurons have thick primary dendrites which are associated with thin lightly immunoreactive processes that give rise to clusters of GAD-positive terminals. Clusters of GAD-positive terminals are prominent in lamina A, A1, magnocellular C, and MIN but are rare in the parvocellular C laminae. Within the A laminae, GAD immunoreactivity is found within vesicle-containing profiles of the synaptic glomerulus lying postsynaptic to optic axon terminals and presynaptic to unlabeled dendritic profiles. GAD-positive neurons in the A laminae are distinguished from other small to medium-sized neurons by their failure to label following injections of HRP into visual cortex and by their lack of cytoplasmic laminated body. These results support the idea that GAD-positive neurons constitute a distinct population of neurons in the lateral geniculate nucleus of the cat; a population which has a number of features in common with previous descriptions of presumed local circuit neurons based on Golgi staining.

Glutamic acid decarboxylase-immunoreactive neurons and horseradish peroxidase-labeled projection neurons in the ventral posterior nucleus of the cat and Galago senegalensis.

Immunocytochemical methods were used to identify neurons in the ventral posterior nucleus of the cat and Galago senegalensis that contain glutamic acid decarboxylase (GAD), the synthetic enzyme for the inhibitory neurotransmitter, GABA. In both species GAD-immunoreactive neurons make up about 30% of the total neurons in the ventral posterior nucleus and form a distinct class of small cells. After cortical injections of horseradish peroxidase (HRP), GAD-immunoreactive cells are not labeled with HRP and may, therefore, be GABAergic local circuit neurons. Comparison of the dendritic morphology of GAD-immunoreactive neurons with that of HRP-filled projection neurons reveals that the morphology of the GAD-containing neurons is distinct and, in particular, that the GAD-immunoreactive neurons display fewer primary dendrites. The relay neurons, in turn, can be divided into classes based on dendritic morphology and cell body size.