Neuronal types in the striatum of man

Summary Nerve cells of the human striatum were investigated with the use of a newly developed technique that reveals the pattern of pigmentation of individual nerve cells by means of transparent Golgi impregnations of their cell bodies and processes. Five types of neurons are distinguished:Type I is a medium-sized spine-laden neuron with an axon giving off a great number of collateral branches. The vast majority of the cells in the striatum belong to this type. Numerous intensely stained lipofuscin granules are contained in one pole of the cell body and may also extend into adjacent portions of a dendrite.Type II is a medium-sized to large neuron with long intertwining dendrites decorated with spines of uncommon shape. A distinguishing feature of this cell type is the presence of somal spines. This cell type is devoid of pigment or contains only a few tiny lipofuscin granules.Type III is a large multipolar neuron. The cell body generates a few rather extended dendrites that are very sparsely spined. The finely granulated pigment is evenly dispersed within a large portion of the cytoplasm.Type IV is a large aspiny neuron with rounded cell body and richly branching tortuous dendrites. The axon branches frequently in the vicinity of the parent soma. Large pigment granules are concentrated within a circumscribed part of the cell body close to the cell membrane.Type V is a small to medium-sized aspiny neuron. The dendrites break up into a swirling mass of thin branches. More than one axon may be given off from the soma. The axons branch close to the soma into terminal twigs. Cells of this type contain numerous large and well-stained lipofuscin granules.Each of the cell types has a characteristic pattern of pigmentation. The different varieties of nerve cells in the striatum can therefore be distinguished not only in Golgi impregnations but also in pigment-Nissl preparations.     

Neuronal types and their percent distribution within the magnocellular nuclei of the human basal forebrain

Three neuronal types constituting the magnocellular nuclei of the human basal forebrain have been differentiated with the aid of preparations stained for both Nissl material and pigment deposits: type I = large multipolar neurons contain loosely packed and faintly stained lipofuscin granules occupying a large portion of the cell body; type II = large spindle-shaped neurons reveal a densely packed accumulation of coarse and intensely stained lipofuscin granules, and type III = small nerve cells, scattered among these large neuronal components, with only a small number of faintly stained lipofuscin granules. The determination of the projection areas of the somata of the three neuronal types has led to a distribution pattern with three peaks. The ratio of the nerve cell types has been evaluated: 73.6% type I; 8.6% type II, and 17.8% type III neurons.     

On the fine structure of the small,heavily pigmented non-pyramidal cells in lamina II and upper lamina III of the human isocortex

Summary With the aid of a newly developed technique for the successive examination of both the Golgi and pigment picture of individual neurons (Braak, 1974a) Braak (1974b) demonstrated that within lamina II and upper lamina III of the human isocortex, heavily pigmented non-pyramidal cells are distributed irregularly and sparsely. The lipofuscin pigment granules serve as excellent internal markers to identify these non-pyramidal cells in ultrathin sections. This favourable circumstance facilitates the study of these interneurons in the electron microscope.The heavily pigmented non-pyramidal cells are small, spherical to ovoid with diameters of about 12–15 m. One pole of the cell comprising a large cytoplasmic area gives rise to a few dendrites, while the other pole is occupied by the nucleus and in some cases is in close apposition to another nerve cell body. The nucleus is deeply invaginated by the large cytoplasmic area and occasionally displays nuclear inclusions. Among the usual organelles distributed within the large cytoplasmic area the mitochondria with a moderately electron dense matrix are abundant and the coarse lipofuscin pigment granules are the most striking elements. The latter contain densely packed filamentous or tubular material and a single vacuole. The perikaryon rarely receives more than 3 type I and type II synapses per section per cell, whereas the dendrites receive numerous synapses of both type I and type II. Within the apposition zone to another nerve cell body (which in no case is a heavily pigmented non-pyramidal cell) puncta adhaerentia occur and also contacts in which the cleft of 8 nm is intersected by a dense stratum.Some of the ultrastructural findings are summarized in the schematic drawing of Figure 15.     

Percentage of projection neurons and various types of interneurons in the human claustrum

Each neuronal type of the human claustrum is differently pigmented. This has been demonstrated by combining a transparent Golgi technique with the specific staining of lipofuscin deposits. One type of projection cells and four varieties of interneurons have been distinguished. In this study the percentage of these different neuronal types has been evaluated using preparations stained for both Nissl material and pigment deposits.     

Morphological characteristics of different types of neurons in the ventrooralis anterior,ventrooralis internus,and ventrooralis posterior nuclei in the human thalamus

1. Golgi-Kopsch preparations of the oral ventral nuclei of human thalamus were analyzed in an attempt to classify the neuronal types. 2. Three types of neurons are described for the first time in humans. Type I neurons are large or medium in size and bear dendrites with protrusions, spines, and short hair-like appendages. Some have a radiate dendritic arbor and others have dendrites grouped in tufts. The dendritic trees of these neurons are dense. 3. Type II neurons are medium or small in size with less dense dendritic trees. These cells have somatic as well as dendritic appendages of different forms. 4. Relatively rare is a type of very small neurons, type III, with few and sparsely branching dendrites.     

Quantitative morphological characteristics of developing neurons of the sensory trigeminal nuclei in kittens

Five types of neurons were distinguished in the sensory nuclei of the trigeminal nerve, stained by Golgi's method, in kittens aged 1–5 days and 30 days: reticular and short-dendritic cells (with few branches), and multipolar giant cells, arborescent, and bushy neurons (densely branching). Yet another special type of cell, with a few short dendrites and one long dendrite, was distinguished in preparations from the brain of newborn kittens. Analysis of the dimensions of the bodies, the number, length, and ramification of the dendrites, and the total ramification of the cell yielded quantitative morphological characteristics of these neurons at different times of development. These types of neurons differed in their qualitative and quantitative parameters and in the features of their maturation.Bushy neurons underwent regressive changes during development. Foci of maximal ramification of dendrites of densely branched neurons changed their location during the first months of life relative to the cell body, moving into the more distal regions of the dendrites. Differences in orientation of dendrites with foci of maximal ramification were found relative to neighboring brain formations, which depended on the types of cells and the animal's age. The high level of maturity of trigeminal neurons at birth was demonstrated.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Brain Institute, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 14, No. 6, pp. 592–600, November–December, 1982.     

AN UNUSUAL CONFIGURATION OF THE GOLGI COMPLEX IN PIGMENT-PRODUCING "TEST" CELLS OF THE OVARY OF THE TUNICATE,STYELA

The test cell in the ovary of the tunicate Styela contains a large and robust Golgi complex which demonstrates a regional structural differentiation. In one of the regions, branching of the lamellae occurs resulting in a honeycomb or lattice-type arrangement. Small, dense granules or homogeneous material of moderate density may be present within certain of the Golgi cisternae. The close association, or continuity in some cases, between elements of the Golgi complex and immature forms of pigment suggests that the Golgi complex in these cells is involved in pigment formation. These relationships are shown and discussed in terms of possible functional significance.     

The pyramidal cells of Betz within the cingulate and precentral gigantopyramidal field in the human brain

Summary It can be demonstrated with the aid of Golgi-, Nissl-, and pigment preparations that the Betz cells represent a homogeneous class of giant cells within the human brain, which can readily be distinguished from other large pyramids by their densely aggregated lipofuscin deposits. In addition to the primary motor field (4, Brodmann), there exists only a small area on the medial surface of the hemisphere in front of the central sulcus which also contains large Betz pyramids in layer Vb. This recently discovered gigantopyramidal field is almost totally buried in the depth of the cingulate sulcus (Braak, 1976b). Compared with the Betz cells of the primary motor field (4, Brodmann), those of the cingulate area display numerous primitive traits. A small number of short basal dendrites springs off from the cell body. The apical dendrite forks in a short distance from the perikaryon repeatedly but issues only few side branches. A spine-free proximal dendritic segment is poorly developed or lacking. Moreover, numerous spines are encountered along the surface of the soma. In view of their primitive features the large pyramids of the cingulate gigantopyramidal area are interpreted as the forerunners of the precentral Betz pyramids.Supported by the Deutsche Forschungsgemeinschaft (Br 317/7). Dedicated to Prof. Dr. med. Drs. h.c. W. Bargmann in honour of this 70th birthday     

Ligh- and electron-microscopic immunocytochemistry of somatotropes in the anterior pituitary gland of European ferret,Mustela putorius furo

Light-microscopic immunocytochemistry of ferret anterior pituitary revealed the localization of somatotropes in the pars distalis, but no immunoreactive cells were detected in the pars tuberalis. Ultrastructural studies by superimposition immunocytochemistry and immuno-electron microscopy, clucidated the morphological heterogeneity of these somatotropic cells. They were classified into 2 subtypes on the basis of size of the secretory granules. Type-I cells with small granules (mean diameter, 192 nm), were considered to be the immature somatotrop, while Type-II cells, with comparatively larger secretory granules (mean diameter, 257 nm), were considered to be the matured form of Type-I cells and the typical somatotropic cell-type, and were much more predominant than the Type-I cells. The fact that Type-II cells had a distinct Golgi zone and many mitochondria, while in Type-I cells the intracellular organelles were generally less developed, supports this suggestion. In addition to these two extreme subtypes, several intermediate forms were also encountered that may represent different transitional phases during the conversion of Type I to Type II. Protein A-gold immuno-electron microscopy illustrated the specific localization of growth hormone over the granules, with no labelling over any other cytoplasmic organelles of the 2 somatotrope subtypes.     

The cytology,histochemistry, and ultrastructure of the cell types found in the digestive gland of the slug,Agriolimax reticulatus (Müller)

Summary Four cell types have been identified in the digestive glands from light and electron microscope studies. The possible functions of each cell type are discussed.Thin cells are undifferentiated. Calcium cells contain spherules of calcium salts which have a characteristic ultrastructure. Different protein granules are found apically. Digestive cells are present as two distinct forms. One form is believed to be absorbing food material and digesting it intracellularly, and the other form is a secreting cell. Both forms contain green and yellow granules and histochemistry shows these granules to be distinct. Protein granules also occur apically.Excretory cells are distinguished by having a large central vacuole containing excretory granules. Histochemistry shows these granules, like the yellow granules of digestive cells, to be composed mainly of lipofuscin.It is suggested that digestive cells form excretory cells.     

A topography and ultrastructural characterization ofin vivo 5,7-dihydroxytryptamine-labeled serotonin-containing neurons in the central nervous system ofAplysia californica

1. Several weeks after administration of 5,7-dihydroxytryptamine (5,7-DHT) to Aplysia, a dark pigmentation appears in serotonin-containing neurons, and this pigmentation allows visual identification of serotonergic neurons but does not appear to alter their physiology. 2. We have determined the distribution of labeled nerve cell bodies in the various ganglia of Aplysia and have characterized the pigment containing structures in both control and labeled neurons. 3. All neurons in this preparation, whether or not they utilize serotonin as a transmitter, contain pigment granules, and three types of pigment granules can be distinguished. After 5,7-DHT a new type of granule appears in serotonergic neurons, probably reflecting lysosomes that have accumulated serotonergic synaptic vesicles that contain the oxidized 5,7-DHT. 4. It remains unclear why this substance does not cause neurotoxicity in mollusks as it does in mammalian preparations.     

Somatostatin-like immunoreactivity in non-pyramidal neurons of the human entorhinal region

Summary The distribution of somatostatin-immunoreactive cells and processes throughout the human entorhinal region and subjacent white matter was examined either by the unlabelled antibody-enzyme method or by the avidin-biotin method. The brain slices were obtained at autopsy with a short post-mortem delay. The majority of somatostatin immunoreactive nerve cells was found in the inner principal layer and subjacent white matter. In addition, individually scattered immunoreactive neurons were observed in both the outer principal layer and lamina dissecans. The immunoreactive perikarya varied in shape and ranged in size from 10 to 30 m. Without exception the neurons could be classified as belonging to the group of non-pyramidal neurons. Each neuron gave rise to a few thick dendrites and a thin axon with a beaded appearance. In the adult human brain, the pattern formed by lipofuscin granules deposited in the nerve cells can be considered characteristic for the type of the neuron. Therefore, immunoreactive perikarya were documented, destained of chromogen and restained to demonstrate lipofuscin pigment and basophilic substance. It became evident from these studies that the previously immunoreactive cells were characterized by a large rounded and eccentrically located nucleus, sparse basophilic substance and, in most cases, a lack of lipofuscin granules. A few of the immunoreactive cells were laden with coarse pigment granules. The findings permit classification of entorhinal somatostatin-immunoreactive neurons as either non-pigmented or pigment-laden non-pyramidal neurons.Dedicated to Prof. Dr. J. Lang, Würzburg, on the occasion of his 65th birthdayA portion of the results has been presented at the annual meeting of the European Neuroscience Association 1986 in Marseille, France (Friederich-Ecsy et al. 1986)     

Neuroglia in the corpus callosum of the primate, slow loris (Nycticebus coucang coucang)

The corpus callosum of adult slow loris consists of a mixed population of several cell types, i.e. free subependymal cells, oligodendrocytes, astrocytes and microglia. The free subependymal cell is rather small and slender with a somewhat patchy nucleus. It shows scanty cytoplasm with free ribosomes. Oligodendrocytes vary both in nuclear and cytoplasmic densities and can be divided into three classes: light, medium dense and dark types. Their cytoplasm contains microtubules, rough endoplasmic reticulum and Golgi saccules. Astrocytes are pale cells with large amount of filaments in their cytoplasm. Microglia are small cells with granulated nuclei. The cells often show large cytoplasmic protrusions containing the usual cell organelles and lipofuscin bodies in their cytoplasm. Lastly, cells with typical features of neurons are occasionally encountered among the white matter.     

The lipofuscin in neuroglial cells of the optic nerve of Formosan rock-monkey (Macaca cyclopis)

The ultrastructure of lipofuscin granules in neuroglial cells of the optic nerve of the Formosan Rock-Monkey was investigated by electron microscopy. In the cytoplasm of astroglial cells, numerous irregular lipofuscin granules were characterized by the presence of large lipid droplets, small electron-dense pigment granules, and some lamellar structures. The lipofuscin granules of the oligodendroglial cells were composed largely of dense, coarse pigment granules, multilinear structures, and a few small lipid droplets. The lipofuscin granules in microglial cells were characterized by numerous lipid droplets in various sizes, small electron-dense pigment granules, and prominent lamellar structures. It was reported that the lipofuscin granules are wear-and-tear materials and products from the cells in lower functional activity. However, our observations suggest that the presence of lipofuscin granules in the neuroglial cells of the optic nerve is likely a characteristic product of active phagocytosis.     

The amygdala of the cat: A golgi study

Summary The amygdaloid complex in the cat was studied in a series of Golgi preparations. Both the lateral and the basal nucleus are composed of the same two cell types, one of which (type P) resembles the pyramidal and the other (type S) the stellate neuron of the cortex. The cortical nucleus can be divided into three layers (I, II, and III–IV) which are made up of cells similar to those in the periamygdaloid cortex. In addition, there are sufficient differences in the organization of these layers to justify a subdivision of the cortical nucleus into lateral and medial parts. The dendrites of neurons in the medial part of the central nucleus, the medial nucleus and the anterior amygdaloid area undergo less branching and carry fewer spines than those of the type P cell. The neurons in the nucleus of the lateral olfactory tract are all of the pyramidal or modified pyramidal type. These findings are discussed in relation to those of previous investigators who employed the Nissl and Golgi methods.This investigation was supported by the Medical Research Council of Canada, Grant M.T. 870. The author wishes to thank Miss Elizabeth Korzeniowski for her technical assistance.     

Shapes of nerve cells in the myenteric plexus of the guinea-pig small intestine revealed by the intracellular injection of dye

Summary The shapes of myenteric neurons in the guineapig small intestine were determined after injecting living neurons with the dye Lucifer yellow via a microelectrode. The cells were fixed and the distribution of Lucifer yellow rendered permanent by an immunohistochemical method. Each of 204 nerve cells was examined in whole-mount preparations of the myenteric plexus and drawn using a camera lucida at 1250 x magnification. Four cell shapes were distinguished: (1) neurons with several long processes corresponding to type II of Dogiel; (2) neurons with a single long process and lamellar dendrites corresponding to type I of Dogiel; (3) neurons with numerous filamentous dendrites; and (4) small neurons with few processes. About 15% of the neurons could not be placed into these classes or into any single class. The type II neurons (39% of the sample) had generally smooth somata and up to 7 (average 3.3) long processes, most of which ran circumferentially. Dogiel type I neurons (34% of sampled neurons) had characteristic lamellar dendrites, i.e., broad dendrites that were flattened in the plane of the plexus. The filamentous neurons (7% of the sample), had, on average, 14 fine processes up to about 50 m in length. Small neurons with smooth outlines and a few fine processes made up 5% of the neurons encountered. We conclude that myenteric neurons that have been injected with dye can be separated into morphologically distinct classes and that the different morphological classes probably correspond to different functional groupings of neurons.     

On the structure of the human striate area. Lamina IVcβ

Summary Layer IVc of the human striate area consists mainly of a great number of small spinous local circuit neurons which store numerous characteristic lipofuscin granules. Since the neurons of the neighbouring layers are almost devoid of pigment deposits the boundaries of lamina IVc are easily traceable. Hence, the pigment granules can be used as internal markers to unequivocally identify these small pigmented spinous local circuit neurons of lamina IVc in ultrathin sections. They have a large spherical nucleus surrounded by a narrow cytoplasmic rim poor in organelles, and very scarcely receive axosomatic symmetric synapses.Within layer IVc four types of synaptic boutons can be distinguished. Type-1-boutons are large, contain a few and loosely arranged round vesicles and make asymmetric synaptic contacts with dendrites and dendritic spines. The type-2-boutons which are also large are filled with densely packed round vesicles which accumulate at the presynaptic membrane. The large type-3-boutons are characterized by elongated vesicles and symmetric synaptic contact zones. These boutons generate several fingerlike protrusions. Small profiles which contain elongated vesicles and form symmetric synaptic contacts, are most probably parts of these protrusions. The large amount of small boutons with round vesicles and asymmetric synaptic contact zones are tentatively described as type-4-boutons although it is far from certain that they represent a uniform class. The presumable origins of the different types of boutons are discussed.Supported by the Deutsche Forschungsgemeinschaft (Br. 634/1)Dedicated to Prof. Dr. med. H. Leonhardt in honor of his 60th birthday     

Retinal pigment epithelium lipofuscin proteomics

Lipofuscin accumulates with age in the retinal pigment epithelium (RPE) in discrete granular organelles and may contribute to age-related macular degeneration. Because previous studies suggest that lipofuscin contains protein that may impact pathogenic mechanisms, we pursued proteomics analysis of lipofuscin. The composition of RPE lipofuscin and its mechanisms of pathogenesis are poorly understood in part because of the heterogeneity of isolated preparations. We purified RPE lipofuscin granules by treatment with proteinase K or SDS and showed by light, confocal, and transmission electron microscopy that the purified granules are free of extragranular material and associated membranes. Crude and purified lipofuscin preparations were quantitatively compared by (i) LC MS/MS proteomics analyses, (ii) immunoanalyses of oxidative protein modifications, (iii) amino acid analysis, (iv) HPLC of bisretinoids, and (v) assaying phototoxicity to RPE cells. From crude lipofuscin preparations 186 proteins were identified, many of which appeared to be modified. In contrast, very little protein ( approximately 2% (w/w) by amino acid analysis) and no identifiable protein were found in the purified granules, which retained full phototoxicity to cultured RPE cells. Our analyses showed that granules in purified and crude lipofuscin preparations exhibit no statistically significant differences in diameter or circularity or in the content of the bisretinoids A2E, isoA2E, and all-trans-retinal dimer-phosphatidylethanolamine. The finding that the purified granules contain minimal protein yet retain phototoxic activity suggests that RPE lipofuscin pathogenesis is largely independent of associated protein. The purified granules also exhibited oxidative protein modifications, including nitrotyrosine generated from reactive nitrogen oxide species and carboxyethylpyrrole and iso[4]levuglandin E(2) adducts generated from reactive lipid fragments. This finding is consistent with previous studies demonstrating RPE lipofuscin to be a potent generator of reactive oxygen species and supports the hypothesis that such species, including reactive fragments from lipids and retinoids, contribute to the mechanisms of RPE lipofuscin pathogenesis.     

Phenotypes of dendritic cells in central lymph of healthy rabbits and during correction of experimental atherosclerosis]

Dendritic cells of central lymph of rabbits have been identified according to the form of the cell body, characteristics of formation and branchiness of its processes in health, in atherosclerosis, its correction with radon, polyphenol preparations made of Sanguisorba officinalis and in combination of the latter. Two main types of dendritic cells have been distinguished. Type I is characterized by a rounded body with clear outlines, protrusions and one compact process. Such cells are often found in lymph of intact animals. Type II has a cell body of various forms with two and more compact or branching processes. This type is mainly detected in atherosclerosis and its correction. The prevalence of the above phenotypes of dendritic cells is attributed to the response of the immune system to atherosclerosis and its correction.