Light and dark Purkinje cells (light microscopic and microautoradiographic studies]

The authors deal with the phenomenon of dualism of pale and dark ganglion cells in general, the phenomenon which, since FLEMMING (1882), still remains an actual problem of neurohistology. They deal with Purkinje cells from a special aspect with the aim to demonstrate the dualism through various staining methods. They directed their attention also to the question of the influence of perfusion and immersion fixation and length of staining with luxol-fast-blue on the production of luxol-positive (chromophilic) Purkinje cells. The authors have found that these methodologic circumstances do not influence the production of luxol-positive Purkinje cells, so it can be hardly spoken about an artifact. Examinations with the labelled leucine have shown that the increased metabolic activity can be observed in those Purkinje cells which in HE-picture are seen as pale ones. In the dark Purkinje cells leucine granulations are located on the surface of plasmatic membrane and they follow to some distance the main dendrite of those cells. The microautoradiographic method evidences for the increased leucine metabolism of the pale Purkinje cells as well.     

Different stainability of Purkinje cells.

Regarding the different stainability when using the Luxol fast blue methods, two kinds of Purkinje cells of the rat are described: Luxol-positive and Luxol-negative cells. Since, by this method, phospholipids are demonstrated, the author suggests the prospective varying functional conditions of these cells. Different tinction of Purkinje cells has been confirmed also by other methods (gallocyanin-chromalaun, thionine, toluidine blue, lithium-haematoxylin, chromalaun-haematoxylin-phloxine and acid phosphatase) in both animal and human material. After 96 h of immobilization the different stainability of Purkinje cells becomes more marked, which penomenon can be as well explained with regard to the functional point of view. Similar differences, though less marked, were found also in neurosecretory cells of the nucelus supra-opticus of the rat and in the nuclear region of the ganglion semilunare Gasseri cells in man. Finally, the author refers to the relations between the Luxol blue staining method and Baker's method employing acid haematoxylin for demonstration of phospholipids in certain kinds of nervous system cells, taking into consideration Kroon's findings.     

Monoclonal antibody to rat brain actin antigenically enhanced with HVJ (Sendai Virus) M protein

Summary Monoclonal antibody to rat brain actin was easily produced using HVJ (Sendai Virus) M protein to enhance the antigenicity of the actin. This monoclonal antibody was determined to be IgM with a kappa light chain. By immunoblot analysis the antibody was also shown to react with rat brain actin but not with HVJ M protein on nitrocellulose sheets. Utilizing the antibody, neuronal cytoplasm in the cerebral cortex, the anterior and posterior horns in the spinal cord, the spinal ganglion and astrocytes showed positive immunohistochemical staining by light microscopy. However, Purkinje cells showed variable staining, some staining intensely, while others were negative. All of neurons in specific anatomical locations showed always positive staining but variable intensities. Vascular walls were stained only faintly. By electron microscopy, neuronal cytoplasm showed diffuse positive staining. Other areas showed a positive reaction, including dendrites, the postsynaptic densities, and a few capillary endothelial cells and arterial smooth muscle cells. The results suggest that the HVJ M protein was effective for producing monoclonal antibody to brain actin, and that the antibody could be utilized for the immunohistochemical study of neuronal elements in both normal and pathological conditions.     

Synaptic inputs to the ganglion cells in the tiger salamander retina

The postsynaptic potentials (PSPs) that form the ganglion cell light response were isolated by polarizing the cell membrane with extrinsic currents while stimulating at either the center or surround of the cell's receptive field. The time-course and receptive field properties of the PSPs were correlated with those of the bipolar and amacrine cells. The tiger salamander retina contains four main types of ganglion cell: "on" center, "off" center, "on-off", and a "hybrid" cell that responds transiently to center, but sustainedly, to surround illumination. The results lead to these inferences. The on-ganglion cell receives excitatory synpatic input from the on bipolars and that synapse is "silent" in the dark. The off-ganglion cell receives excitatory synaptic input from the off bipolars with this synapse tonically active in the dark. The on-off and hybrid ganglion cells receive a transient excitatory input with narrow receptive field, not simply correlated with the activity of any presynaptic cell. All cell types receive a broad field transient inhibitory input, which apparently originates in the transient amacrine cells. Thus, most, but not all, ganglion cell responses can be explained in terms of synaptic inputs from bipolar and amacrine cells, integrated at the ganglion cell membrane.     

Monoclonal antibody to rat brain actin antigenically enhanced with HVJ (Sendai virus) M protein

Monoclonal antibody to rat brain actin was easily produced using HVJ (Sendai Virus) M protein to enhance the antigenicity of the actin. This monoclonal antibody was determined to be IgM with a kappa light chain. By immunoblot analysis the antibody was also shown to react with rat brain actin but not with HVJ M protein on nitrocellulose sheets. Utilizing the antibody, neuronal cytoplasm in the cerebral cortex, the anterior and posterior horns in the spinal cord, the spinal ganglion and astrocytes showed positive immunohistochemical staining by light microscopy. However, Purkinje cells showed variable staining, some staining intensely, while others were negative. All of neurons in specific anatomical locations showed always positive staining but variable intensities. Vascular walls were stained only faintly. By electron microscopy, neuronal cytoplasm showed diffuse positive staining. Other areas showed a positive reaction, including dendrites, the postsynaptic densities, and a few capillary endothelial cells and arterial smooth muscle cells. The results suggest that the HVJ M protein was effective for producing monoclonal antibody to brain actin, and that the antibody could be utilized for the immunohistochemical study of neuronal elements in both normal and pathological conditions.     

鲫鱼视网膜锌离子分布的光、电镜观察

本文应用ｎｅｏ－Ｔｉｍｍ染色法,观察了鲫鱼视网膜内锌离子的分布情况以及明,暗适应条件下鲫鱼视网膜内锌离子分布的变化。结果发现,明适应条件下,外网层、部分光感受器、双极细胞、无长突细胞以及神经节细胞胞体锌离子着色明显,含锌光感受器和双极细胞的突起伸入外网层,暗适应条件下,外网层锌离子染色减弱或消失（Ｐ〈０．０１）。外核层胞体锌离子染色阴性,少数散在分布的视锥细胞呈锌离子阳性,上述资料提示,明适应条件     

Biological rhythms in the context of light at night (LAN)

In mammals including man, the most important zeitgeber for endogenous rhythms is the environmental light/dark cycle. Mammals perceive light through the eyes and that perception is relayed to the suprachiasmatic nucleus (SCN) by means of neuronal signals. The SCN, in turn, innervates the pineal gland, resulting in the production and release of melatonin almost exclusively during night-time hours. Thus, besides object recognition, eyes serve as the sensory organ for detecting the presence or absence of light. The way that light entrains the SCN is still a matter of intense research. It has been shown, for example, that the light intensities required for affecting melatonin rhythms are much higher than the intensities needed for object identification. On the other hand, even in rodents who completely lack the "classical" photoreceptors of the retina, their endogenous rhythms still can be synchronized by normal light/dark cycles. These two observations led to the hypothesis that there must be photoreceptors, apart from the known (object-identifying) retinal photoreceptors, which are responsible for the entrainment of internal rhythms. Very recently, a number of reports showed that in fact a completely new type of retinal photoreceptor, located in ganglion cells, may be responsible for entraining the SCN. It contains a photopigment, melanopsin, which shares homologies with rhodopsin, but also is evolutionarily older. Compared to rods or cones, the melanopsin-containing neurons are rare, but evenly distributed within the retina, indicating that they serve as a global, integrating light sensor. These ganglion cells apparently project directly into the SCN. Taken together, these new developments in photo-chronobiology open new areas of research. It will be of special interest, for example, to determine how the photosensitive ganglion cells and their dendrites integrate the environmental light stimuli.     

Ultrastructural architecture of pulmonary small-granule cell clusters in adult Syrian golden hamster

Throughout the epithelial lining of the respiratory system is a class of cells with characteristics similar to Amine Precursor Uptake and Decarboxylation (APUD) polypeptide hormone-producing cells. In the intrapulmonary airways, these small-granule cells (SGCs) occur either singly or in organized clusters. Although no specific peptide has yet been identified, subclasses have been postulated based on granule geometry or light microscopic staining. The present study characterizes the architectonic and cellular organization of clustered SGCs in the adult Syrian golden hamster. Two morphologically distinct cells can be defined in such clusters, "light" and "dark." Thid distinction was based primarily on differences in the electron density of the cytoplasmic matrix rather than on the remarkable variations in cellular organelles or dense-core secretory vesicles. Both cell types were normal as judged by uniform spherical nuclei, chromatin organization, and distribution of cellular organelles. The "dark" cells, however, presented the profile of a cell actively involved in synthesis with a markedly dilated perinuclear cisterna and endoplasmic reticulum. Additionally, the "dark" cells contained membrane-delimited structures containing concentric membranous whorls, clear vacuoles, and lipofuscin granules. Occasionally, cells were observed to contain features of both cell types, suggesting that they may represent a continuum of common cell lineage. Accordingly, in the absence of additional morphologic or biochemical data, the "light" and "dark" cells most probably correspond to different stages of functional activity or age-related changes of a single type of cell. Unmyelinated nerve endings were occasionally interposed between cells, but synaptic specializations were not observed. Beneath the clusters, nerve fibers were also present, but they were never observed to penetrate the basal lamina or contact any of the SGCs. Of equal occurrence were elements of the vascular system and smooth muscle, suggesting that some SGCs in the adult hamster may function in a paracrine or endocrine manner. Such knowledge is essential to any study attempting to delineate the functional role or roles of these enigmatic organoids.     

Simplified staining in situ of neurosecretory cells of the supraoesophageal ganglion of the female Culex pipiens molestus (Forskal) (Diptera: Culicidae)

The author describes the morphology and distribution of the neurosecretory cells in the supraoesophageal ganglion of the adult female Culex pipiens molestus, using paraldehyde fuchsin and paraldehyde thionine-paraldehyde fuchsin as vital staining techniques. The brain of Culex pipiens molestus has three regions (the proto-, deuto- and tritocerebrum) in which principally two types of neurosecretory cells (A and B) can be detected. Both A (dark) and B (light) cells are to be found in the protocerebrum, where they are termed medial neurosecretory cells, as they are distributed in the pars intercerebralis and only a few occur more laterally. A small group of type A and B neurosecretory cells is to be found in the posterior part of the brain (the tritocerebrum). These cells display characteristics differences in their tinctorial affinity.     

胚胎与成人三叉神经节细胞形态对比观察

目的：探讨胚胎与成人三叉神经节细胞的形态学特征。方法：采用常规HE染色,运用电子显微超微结构技术及特殊染色,对比观察二者的形态学特征。结果：胚胎与成人三叉神经节细胞大体分布及形态一致,与胚胎三叉神经节细胞相比,成人最突出的特点是节细胞大,核仁清晰,居中。电镜下节细胞的细胞器较丰富,发育完善。且成人三又神经节神细胞明暗区别明显。结论t三叉神经节细胞结构与功能完善是在成年之后,成人三又神经节细胞存在明暗两类细胞。     

Immunocytochemical localization of a chondroitin sulfate proteoglycan in nervous tissue. I. Adult brain, retina, and peripheral nerve

Monospecific antibodies were prepared to a previously characterized chondroitin sulfate proteoglycan of brain and used in conjunction with the peroxidase-antiperoxidase technique to localize the proteoglycan by immunoelectron microscopy. The proteoglycan was found to be exclusively intracellular in adult cerebellum, cerebrum, brain stem, and spinal cord. Some neurons and astrocytes (including Golgi epithelial cells and Bergmann fibers) showed strong cytoplasmic staining. Although in the central nervous system there was heavy axoplasmic staining of many myelinated and unmyelinated fibers, not all axons stained. Staining was also seen in retinal neurons and glia (ganglion cells, horizontal cells, and Muller cells), but several central nervous tissue elements were consistently unstained, including Purkinje cells, oligodendrocytes, myelin, optic nerve axons, nerve endings, and synaptic vesicles. In sympathetic ganglion and peripheral nerve there was no staining of neuronal cell bodies, axons, myelin, or Schwann cells, but in sciatic nerve the Schwann cell basal lamina was stained, as was the extracellular matrix surrounding collagen fibrils. Staining was also observed in connective tissue surrounding the trachea and in the lacunae of tracheal hyaline cartilage. These findings are consistent with immunochemical studies demonstrating that antibodies to the chondroitin sulfate proteoglycan of brain also cross-react to various degrees with certain connective tissue proteoglycans.     

The chicken trigeminal ganglion. I. An anatomical analysis of the neuron types in the adult

An anatomical analysis of the chicken trigeminal ganglion was made using light microscopy on specimens prepared by usual chemical fixation or freeze-drying methods and by electron microscopy. Two types of neurons were consistently seen, dark and light cells. Dark cells contained a dense cytoplasm with Nissl substance distributed evenly throughout, whereas light cells had a less dense cytoplasm containing clumps of Nissl substance. The Nissl bodies in light cells contained only a few small cisternae of granular endoplasmic reticulum as compared with many stacked cisternae in Nissl bodies of dark cells. The ratio of dark to light cells was approximately 62:38 in all regions of the ganglion. Dark cells were consistently smaller than light cells. In the seven-day old chick, the mean diameters of the dark and light neurons were 21.4 μ and 29.5 μ respectively; in the adult the values were 29.9 μ and 39.7 μ respectively. It is concluded that the dark and light cells belong to two distinct neuronal cell populations.     

Light Exposue Activates Retina Ganglion Cell Lysophosphatidic Acid Acyl Transferase and Phosphatidic Acid Phosphatase by a c-fos-Dependent Mechanism

We previously reported that the biosynthesis of phospholipids in the avian retina is altered by light stimulation, increasing significantly in ganglion cells in light and in photoreceptor cells in dark. In the present work, we have determined that light significantly increases the incorporation of [3H]glycerol into retina ganglion cell glycerophospholipids in vivo by a Fos-dependent mechanism because an oligonucleotide antisense to c-fos mRNA substantially blocked the light-dark differences. We also studied in vitro the enzyme activities of phosphatidate phosphohydrolase (PAPase), lysophosphatidate acyl transferase (AT II), and phosphatidylserine synthase from retinas of chickens exposed to light or dark. Higher PAPase I and AT II activities were found in incubations of retinal ganglion cells from animals exposed to light; no increase was observed in preparations obtained from light-exposed animals treated with the c-fos antisense oligonucleotide. No light-dark differences were found in phosphatidylserine synthase activity. These findings support the idea that a coordinated photic regulation of PAPase I and AT II is taking place in retina ganglion cells. This constitutes a reasonable mechanism to obtain an overall increased synthesis of glycerophospholipids in stimulated cells that is mediated by the expression of Fos-like proteins.     

胚胎与成人三叉神经节细胞形态对比观察*

摘要目的：探讨胚胎与成人三叉神经节细胞的形态学特征。方法：采用常规HE 染色,运用电子显微超微结构技术及特殊染色,对 比观察二者的形态学特征。结果：胚胎与成人三叉神经节细胞大体分布及形态一致,与胚胎三叉神经节细胞相比,成人最突出的 特点是节细胞大,核仁清晰,居中。电镜下节细胞的细胞器较丰富,发育完善。且成人三叉神经节神细胞明暗区别明显。结论：三 叉神经节细胞结构与功能完善是在成年之后,成人三叉神经节细胞存在明暗两类细胞。     

Fluctuations in amino acid-H3 incorporation into the proteins of the isolated retina]

The rate of amino acid incorporation into the mouse retina proteins was measured in dynamics of dark adaptation and at different terms of their stimulation by light. It was found that variations in dry weight of the ganglion retinal cells and in the incorporation of amino acids into their proteins occurred rhythmically, with the period of about one hour.     

Changes in the cell population of taste buds during degeneration and regeneration of their sensory innervation

Summary Taste buds of rabbit foliate papillae were observed in control, after denervation and during reinnervation by the glossopharyngeal nerve. In control, taste bud cells could be divided into three groups according to their shapes and staining characteristics. Most of the cells were identified as either dark (corresponding to gustatory) or light (corresponding to supporting) cells. However, some cells were encountered which could not readily be placed in either group; they have been termed intermediate cells. Nine to twelve hours after axotomy, wandering cells were observed in many of the taste buds. Thereafter taste buds gradually decreased in size and disappeared, for the most part, by the 14th postoperative day. It was found that dark cells disappeared first, then at a later stage the light cells also disappeared. During reinnervation, dark cells were first to appear about 40 days after the operation and light cells were not seen till about 9 days later.From the observations, it is concluded that the dark cells of the taste bud differentiate from epithelial cells under the influence of nerves and mature into light cells through intermediate cells.     

Autoradiographic research on the dark and light Purkinje cells in the cerebellum of rats

Intensity of the 3H-sodium acetate and 3H-leucine incorporation into dark and light Purkinje cells of the rat cerebellum was studied. The intensity of incorporation into light Purkinje cells was found to be 1.5 times higher than into the dark ones.     

Heterogeneity in the myoglobin content of chicken heart Purkinje fibers

Summary The myoglobin content of chicken myocardial cells was studied using indirect-immunoperoxidase histochemistry. While ordinary myocardial cells exhibited a homogeneous reaction pattern, the reactions for ventricular Purkinje fibers were remarkably heterogeneous. On the basis of the degree of staining, three types of cells, i.e., dark, intermediate, and clear, were distinguishable. In addition, the cytological heterogeneity of Purkinje cells was confirmed using conventional and immunological electron microscopy. The dark cells contained more myofibrils, mitochondria, and other organelles (e.g., ribosomes) than the clear cells.A part of this study was supported by a scientific research grant (59440037) from the Ministry of Education, Japan     

The demonstration of phospholipids in certain cells in neurosecretory nuclei in the rat with Baker's acid haematein after fixation in glutaraldehyde-formol-calcium

Summary With Baker's acid haematein test certain ganglion cells in the brain, their processes and, at some sites, glial cells around blood vessels stain dark blue. This article describes a study of the Baker-positive cells which occur in and around the neurosecretory nuclei. By substituting formol-calcium fixation with glutaraldehyde-formol-calcium fixation shrinkage in brain tissue is completely avoided. If such fixation is used the argument that positive staining of ganglion cells with Baker's method only indicates that these are shrunken neurons can no longer be maintained. A comparative histological study, especially of Baker's technique and controlled chromation (Elftman) showed that the Baker-positive cells contain a phospholipid, probably bound to a protein, as a labile compound, which is easily lost. We found that to immobilize and localize this labile compound in the ganglion cells the technique of fixation and the pH during chromation (which should be around 3.8) are of fundamental importance. Only under these conditions is the complex sufficiently immobilized to allow of its demonstration with acid haematein. These requirements are now completely met if Baker's acid haematein technique is used. The article stresses that only prefixed and chromated frozen sections can be used for this method, thus avoiding shrinkage and non-specific staining of proteins. The modified Baker method as used by us gives constant and reproducible staining and is described in this article. The functional significance of the Baker-positive reaction in some ganglion cells in the n. s. nuclei or glial cells around blood vessels is not dealt with in this article.     

Change in the staining properties of pyriform neurocytes in the mouse cerebellum during protein-calorie deficiency and subsequent rehabilitation

Quantitative analysis has been carried out on semithin sections of cerebellum cortex to investigate the relation between Purkinje cells with different dyeing properties. The number of dark Purkinje cells was found to increase after a month-long food rehabilitation of ill-fed mice. At the same time addition of carnitine to the mouse food has resulted in a significant decline in the number of dark Purkinje cells, as compared to control animals. The data obtained suggest that the rising number of dark Purkinje cells in the cerebellum cortex under conditions of malnutrition is probably due to the increased intracellular accumulation of free fatty acids.