The histopathology of hypothalamic hamartomas: study of 57 cases.

Hypothalamic hamartomas (HHs) are rare developmental tumors that cause seizures or pituitary axis dysfunction, usually beginning in childhood. We analyzed HH tissue from 57 patients whose tumors were resected through recently developed transcallosal interforniceal and transventricular endoscopic surgical approaches. All cases were composed of abnormally distributed but cytologically normal neurons and glia, including fibrillary astrocytes and oligodendrocytes. Neuronal elements predominated in most cases, but a relative increase in astrocytic elements was seen with increasing age. All had various sized nodular foci of neurons as well as areas of diffusely distributed neurons with interspersed glial cells. Smaller neurons predominated, and most cases had only a few interspersed large ganglion cells. Immunohistochemistry demonstrated extensive production of synapse-associated proteins. Immunohistochemistry for phosphorylated and nonphosphorylated neurofilament and alpha-internexin demonstrated staining patterns consistent with mature neurons. In contrast to cortical dysplasia, atypical large ganglion-like balloon cells were almost never seen. In summary, although their number and distribution vary, mature smaller neurons were the most prominent and most consistent histologic feature of HH. Nodules of these small neurons were a universal feature of the microarchitecture of HH lesions associated with epilepsy. Characterization of these neurons may aid in understanding the mechanism of seizure development in HH.

Honeycomb-like mosaic at the border of layers 1 and 2 in the cerebral cortex.

In this report, we present evidence of a small-scale modularity (<100 microm) at the border of layers 1 and 2 in neocortical areas. The modularity is best seen in tangential sections, with double-labeling immunohistochemistry to reveal overlapping or complementary relationships of different markers. The pattern is overall like a reticulum or mosaic but is described as a "honeycomb," in which the walls and hollows are composed of distinct afferent and dendritic systems. We demonstrate the main components of the honeycomb in rat visual cortex. These are as follows: (1) zinc-enriched, corticocortical terminations in the walls, and in the hollows, thalamocortical terminations (labeled by antibody against vesicular glutamate transporter 2 and by cytochrome oxidase); (2) parvalbumin-dense neuropil in the walls that partly colocalizes with elevated levels of glutamate receptors 2/3, NMDAR receptor 1, and calbindin; and (3) dendritic subpopulations preferentially situated within the walls (dendrites of layer 2 neurons) or hollows (dendrites of deeper neurons in layers 3 and 5). Because the micromodularity is restricted to layers 2 and 1b, without extending into layer 3, this may be another indication of a laminar-specific substructure at different spatial scales within cortical columns. The suggestion is that corticocortical and thalamocortical terminations constitute parallel circuits at the level of layer 2, where they are segregated in association with distinct dendritic systems. Results from parvalbumin staining show that the honeycomb mosaic is not limited to rat visual cortex but can be recognized at the layer 1-2 border in other areas and species.

Ontogenetic changes in neuropeptide Y-immunoreactive cerebrospinal fluid-contacting neurons in the hypothalamus of the cloudy dogfish, Scyliorhinus torazame (Elasmobranchii).

Ontogenetic changes in neuropeptide Y-immunoreactive (NPY-ir) cerebrospinal fluid (CSF)-contacting neurons in the dogfish hypothalamus were studied immunohistochemically. NPY-ir CSF-contacting neurons first appeared in the median infundibular floor of the embryo at the 34 mm stage. At the 40 mm stage, similar neurons were found also in the saccus vasculosus (SV). The number of these neurons increased during the 54-80 mm stages, and the cells in the infundibular floor extended their basal processes to the neuropil of the median eminence, whereas the cells in the SV sent their axonal fibers to the tractus sacci vasculosi. After hatching, NPY immunoreactivity in the ventral hypothalamus became less dense, and the labeled CSF-contacting neurons tended to be confined to the nucleus lateralis tuberis, similarly as in the adults. The occurrence of NPY-ir CSF-contacting neurons in the SV was transient during the embryonic periods.

Neural pathways connecting the deutocerebrum and lateral protocerebrum in the brains of decapod crustaceans.

The olfactory and accessory lobes of eureptantian decapod crustaceans are bilateral brain neuropil regions located within the deutocerebrum. Although the olfactory lobe seems to receive only primary olfactory inputs, the accessory lobe receives higher-order multimodal (including olfactory) inputs. The output pathways from both the olfactory and accessory lobes are provided by the axons of a large population of projection neurons, whose somata lie adjacent to the lobes. The axons of these neurons form a large tract that projects bilaterally to the medulla terminalis and hemiellipsoid body in the lateral protocerebrum. To gain insights into the ways in which olfactory information is processed on leaving the deutocerebrum, we examined the neuroanatomy of the projection neuron pathways of three species of eureptantian decapod crustaceans: the freshwater crayfish, Procambarus clarkii and Orconectes rusticus, and the clawed lobster, Homarus americanus. Projection neurons were labeled by focal injections of the lipophilic tracers DiI and DiA into the olfactory and accessory lobes. In all three species, projection neurons innervating the accessory lobe were found to exclusively innervate the neuropils of the hemiellipsoid body. In contrast, projection neurons innervating the olfactory lobes primarily target neuropil regions of the medulla terminalis. The results of this study indicate, therefore, that the projection neuron pathways from the olfactory and accessory lobes project to separate, largely nonoverlapping regions of the lateral protocerebrum. The implications of these findings for our understanding of the processing of olfactory information in the brains of decapod crustaceans are discussed.

Development and connectivity of olfactory pathways in the brain of the lobster Homarus americanus.

The main output pathways from the olfactory lobes (primary olfactory centers) and accessory lobes (higher-order integrative areas) of decapod crustaceans terminate within both of the main neuropil regions of the lateral protocerebrum: the medulla terminalis and the hemiellipsoid body. The present study examines the morphogenesis of the lateral protocerebral neuropils of the lobster, Homarus americanus, and the development of their neuronal connections with the paired olfactory and accessory lobes. The medulla terminalis was found to emerge during the initial stages of embryogenesis and to be the target neuropil of the output pathway from the olfactory lobe. In contrast, the hemiellipsoid body is first apparent during mid-embryonic development and is innervated by the output pathway from the accessory lobe. The dye injections used to elucidate these pathways also resulted in the labeling of a previously undescribed pathway linking the olfactory lobe and the ventral nerve cord. To increase our understanding of the morphology of the olfactory pathways in H. americanus we also examined the connectivity of the lateral protocerebral neuropils of embryonic lobsters. These studies identified several interneuronal populations that may be involved in the higher-order processing of olfactory inputs. In addition, we examined the neuroanatomy of ascending pathways from the antenna II and lateral antenna I neuropils (neuropils involved in the processing of chemosensory and tactile inputs). These studies showed that the ascending pathways from these neuropils innervate the same regions of the medulla terminalis and that these regions are different from those innervated by the olfactory lobe output pathway.

Distribution of GABA(B) receptor protein in somatosensory cortex and thalamus of adult rats and during postnatal development.

In the present study we report the immunolocalisation of gamma-aminobutyric acid (GABA)(B) receptors within the cerebral somatosensory cortex (S1) and thalamus of adult and young (1-22 postnatal days) rats. The antibody used recognises a peptide in the carboxy-terminal domain and therefore did not distinguish between the different isoforms GABA(B)1a or GABA(B)1b. The results showed that GABA(B) receptor protein was widely distributed in the brain of both adult and young rats, with different degrees of labelling in separate cerebral nuclei. Antibody labelling was localised both on cells and the neuropil. In the cerebral cortex of adult animals the highest immunolabelling was evident in layers V and VIb, although immunoreactivity was also present in the superficial layers. The strongest signal was evident in the medial habenula.The thalamus showed labelling in the reticular, ventrobasal and geniculate nuclei. In the first postnatal days GABA(B) expression was evident in the cortical cells of layer V, VIb and in the cortical plate. The pattern of labelling in the cerebral cortex of young rats became indistinguishable from that of adult rats by day 12. In the thalamus, the main difference compared to the adult pattern was observed in the mediodorsal nucleus which, in early development, showed a high immunosignal, however, by postnatal day 22 the immunoreactivity decreased with only some scattered cells labelled in the adult brain.

A new look at embryonic development of the visual system in decapod crustaceans: neuropil formation, neurogenesis, and apoptotic cell death.

In recent years, comparing the structure and development of the central nervous system in crustaceans has provided new insights into the phylogenetic relationships of arthropods. Furthermore, the structural evolution of the compound eyes and optic ganglia of adult arthropods has been discussed, but it was not possible to compare the ontogeny of arthropod visual systems, owing to the lack of data on species other than insects. In the present report, we studied the development of the crustacean visual system by examining neurogenesis, neuropil formation, and apoptotic cell death in embryos of the American lobster, Homarus americanus, the spider crab, Hyas araneus, and the caridean shrimp, Palaemonetes argentinus, and compare these processes with those found in insects. Our results on the patterns of stem cell proliferation provide evidence that in decapod crustaceans and hemimetabolous insects, there exist considerable similarities in the mechanisms by which accretion of the compound eyes and growth of the optic lobes is achieved, suggesting an evolutionary conservation of these mechanisms.

Molecular cloning of a lobster Gbeta subunit and Gbeta expression in olfactory receptor neuron dendrites and brain neuropil.

We have isolated from the olfactory organ of the American lobster (Homarus americanus) two cDNA clones with homology to beta subunits of G proteins. LobGbeta1 contained a complete open reading frame that predicted an amino acid sequence with >80% identity to Gbeta sequences from other species. LobGbeta2 was a fragment of an open reading frame whose predicted amino acid sequence had 65-69% identity to other Gbeta sequences. LobGbeta2 mRNA was not detectable in the brain, eye plus eyestalk, leg, dactyl, olfactory organ, or tail muscle. In contrast, lobGbeta1 was expressed in all these tissues as a single mRNA species of 6.4 kb and a protein of 37 kD. In the brain and olfactory organ, Gbeta immunoreactivity was almost exclusively confined to neurites: the neuropil regions of the brain and the outer dendrites of the olfactory receptor neurons. Coimmunoprecipitation revealed that lobster Gbeta interacted with both Galpha s and Galpha q. LobGbeta1 is likely to be involved in a wide range of signaling events including olfactory transduction and synaptic transmission in the brain.

Morphological organization of neuropile glial cells in the central nervous system of the medicinal leech (Hirudo medicinalis).

Neuropile glial (NPG) cells in the central nervous system of the medicinal leech, Hirudo medicinalis, were studied by histological, histochemical and immunocytochemical techniques. The NPG cells are often surrounded by electron-dense microglial cells. The central cytoplasm of NPG cells shows a significant zonation. The zone around the nucleus contains mitochondria, glycogen and vesicles. The cytoplasm also contains many ribosomes, a few dictyosomes and distinct inclusions up to 2 microns in diameter. A second zone around the perinuclear region is marked by the occurrence of bundles of intermediate filaments that correspond in thickness to glial filaments of vertebrates. We found a positive reaction with polyclonal antibodies against human glial fibrillary acidic protein (GFAP), and the areas of intense fluorescence correspond to the regions where intermediate filaments were found to be abundant. The peripheral zone contains numerous membrane stacks that could not be contrasted by lanthane nitrate or tannic acid. Therefore, the membrane stacks could be part of an extensive smooth endoplasmic reticulum, which is characteristic of cells with active lipid metabolism.