Cytologic characteristics of "dark" and "light" cells in the brain amygdaloid complex

This paper reports data on cytological peculiarities of neurons of two main zones of sexual dimorphism in brain amygdala (dorsomedial nucleus and anterior cortical nucleus). The main attention was paid to some characteristics of "dark" and "pale" cells found in the amygdaloid complex for the first time. It is supposed that the dark and pale cells are targets for gonadal steroids, whose cyclic changes in concentration in the blood difined their functional states. Though the ultrastructure of dark and pale cells of the amygdaloid complex is similar to that of neurosecretory cells of hypothalamus, there are necessary electron microscopic and cytochemical evidences.     

Electron microscopic-cytochemical study of the enzyme activity of energy metabolism in "dark" and "light" neurons of the rat cerebral cortex

The ultrastructural localization of succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH) activity in "dark" and "light" neurons of the intact rat's frontal brain cortex has been studied. The enzymes' activity was detected with using potassium ferricyanide as artificial acceptor of electrons. In the "light" cells SDH activity is localized in the mitochondria and plasma membranes. LDH activity is localized in the mitochondria, plasma membranes and hyaloplasm. SDH and LDH activity was not found in the "dark" cells.     

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.     

Neuronal response in the CA1 field of the cat dorsal hippocampus to visual stimuli

Responses of 46 neurons of the CA₁ field, of the dorsal hippocampus to visual stimuli were investigated during acute experiments on awake cats following pretrigeminal brainstem action. The receptive field was small in size in 71% of hippocampal neurons. The cells responded both tonically (34%) and phasically (66%) to the presentation of immobile stimuli. All the test cells of the CA₁ field of the dorsal hippocampus responded to moving visual stimuli and 27% of these neurons were directionally tuned. A group of 7% of the neurons displayed particular sensitivity to the movement of a dark spot across the receptive field; these cells frequently reacted more to a moving dark spot than to a bar. Findings indicate the presence of highly specific sensory neurons within the hippocampus.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 17, No. 6, pp. 779–786, November–December, 1985.     

Gel-to-gel phase transition may occur in mammalian cells: Mechanism of formation of "dark" (compacted) neurons

In the course of many diseases, individual non-apoptotic cells that are randomly distributed among undamaged cells in various mammalian tissues become shrunken and hyperbasophilic ("dark"). The light microscopic shrinkage is caused by a potentially reversible, dramatic compaction of all ultrastructural elements inside the affected cells, and escape of the excess water through apparently intact plasma membrane. In the case of neurons, the ultrastructural compaction rapidly involves the soma-dendrite domains in an all-or-nothing manner, and also mm-long axon segments. The present paper demonstrates that such ultrastructural compaction in neurons, which affects the whole soma-dendrite domain or long axon segments, can take place both immediately after an in vivo head injury and in rat brains perfusion-fixed for 30 min., and then chilled to just above the freezing point before the same kind of head injury was inflicted. This argues strongly against any enzyme-mediated compaction mechanism. On the analogy of gel-to-gel phase transitions in polymer chemistry, we hypothesize a pure physico-chemical compaction mechanism. Specifically, after initiation at a single site in each affected cell, the ultrastructural compaction is propelled throughout the whole cell on the domino principle by the free energy stored in the form of non-covalent interactions among the constituents of some cytoplasmic gel structure.     

Transient Morphological Alterations in the Hippocampus After Pentylenetetrazole-Induced Seizures in Rats

The relationships between seizures, neuronal death, and epilepsy remain one of the most disputed questions in translational neuroscience. Although it is broadly accepted that prolonged and repeated seizures cause neuronal death and epileptogenesis, whether brief seizures can produce a mild but similar effect is controversial. In the present work, using a rat pentylenetetrazole (PTZ) model of seizures, we evaluated how a single episode of clonic–tonic seizures affected the viability of neurons in the hippocampus, the area of the brain most vulnerable to seizures, and morphological changes in the hippocampus up to 1 week after PTZ treatment (recovery period). The main findings of the study were: (1) PTZ-induced seizures caused the transient appearance of massively shrunken, hyperbasophilic, and hyperelectrondense (dark) cells but did not lead to detectable neuronal cell loss. These dark neurons were alive, suggesting that they could cope with seizure-related dysfunction. (2) Neuronal and biochemical alterations following seizures were observed for at least 1 week. The temporal dynamics of the appearance and disappearance of dark neurons differed in different zones of the hippocampus. (3) The numbers of cells with structural and functional abnormalities in the hippocampus after PTZ-induced seizures decreased in the following order: CA1?>?CA3b,c?>?hilus?>?dentate gyrus. Neurons in the CA3a subarea were most resistant to PTZ-induced seizures. These results suggest that even a single seizure episode is a potent stressor of hippocampal neurons and that it can trigger complex neuroplastic changes in the hippocampus.     

Photoperiodic response of serotonin- and galanin-immunoreactive neurons of the paraventricular organ and infundibular nucleus in Japanese quail, Coturnix coturnix japonica

We investigated the photoperiodic response of serotonin- and galanin (GA)- immunoreactive (ir) cells in the paraventricular organ (PVO) and infundibular nucleus (IF) of the Japanese quail and the interaction of these cells with gonadotropin-releasing hormone (GnRH)-ir neurons in the hypothalamus. Serotonin-ir cells were located in series from the PVO to the IF, and were connected with each other. The number of serotonin-ir cells differed significantly between light and dark phases on the short days (SD), but did not differ between light and dark phases on long days (LD). GA-ir cells were also found in the PVO and IF. The number of GA-ir cells under SD conditions was significantly greater than under LD conditions but did not change diurnally. Both serotonin-ir and GA-ir fibers ran along the GnRH-ir cells in the nucleus commissurae pallii. Serotonin-ir and GA-ir fibers were connected with the GnRH-ir fibers in the external layer of the median eminence (ME). We confirmed that GA-ir fibers were closely associated with serotonin-ir neurons in the PVO and IF. GA-ir neurons have at least 2 routes of regulating GnRH neurons directly, and indirectly via the serotonin-ir cells in the PVO and IF.     

Morphological effects of transcardially perfused SDS on the rat brain

Background information. For explanation of the formation of ‘dark’ neurons, an enigmatic phenomenon in neuropathology, we hypothesized recently that all spaces between the ultrastructural elements visible in the traditional transmission electron microscope are filled with a gel structure that stores free energy in the form of non‐covalent interactions, is continuous in the whole soma—dendrite domains of neurons, and is capable of whole‐cell phase transition. This hypothesis was deduced from the fact that ‘dark’ neurons can be formed, even under conditions extremely unfavourable for enzyme‐mediated biochemical processes, if initiated by a physical damage. In order to gain further information on this gel structure, we perfused transcardially rats for 5 min with physiological saline containing 1 mM SDS before the perfusion of a fixative for electron microscopy. Results. Dramatic compaction of visibly intact ultrastructural elements was caused in the whole soma—dendrite domains of thinly scattered neurons (‘dark’ neurons), whereas substantial cytoplasmic swelling and patchy ultrastructural disintegration occurred in numerous other neurons (‘light’ neurons). Similar morphological changes were observed in scattered astrocytes, oligodendrocytes, pericytes and endothelial cells. Conclusions. These observations: (i) support the existence of the above intracellular gel structure in neurons; (ii) allow the conclusion that this gel structure is present in the form of an ubiquitous trabecular network surrounded by a confluent system of fluid cytoplasm; (iii) draw attention to the possibility that the previous two statements also apply to other cell types of the brain tissue; and (iv) suggest that pressure‐induced direct channels exist between neurons and astrocytes.     

Polarization microscopic evidence for an oriented cytoplasmic structure in the "dark" variants of adrenalin-storing cells.

A diffuse cytoplasmic birefringence confined to "dark" adrenalinstoring cells has been described. The main optical characteristics of the birefringence factor include: regular orientation of birefringence relative to the base-apex axis of cells; additive anisotropic staining with methods based on the principle of topo-optical staining reactions; dependence of birefringence on labile morphologic properties. On the basis of the capacity of the macromolecular matrix of chromaffin granules to form lamellar liposomal structures in vitro it has been proposed that a reorientation of molecular organization in the matrix of chromaffin cells is responsible for the observed optical phenomenon. The direction of birefringence was explained by a preferential direction of contractile forces acting during "dark" cell formation.     

Influence of continuous light and darkness on the secretory pinealocytes of<Emphasis Type="Italic">Heteropneustes fossilis</Emphasis>

In an earlier study onHeteropneustes fossilis, evidence of secretory activity in the pinealocytes had been demonstrated at the electron microscopic (EM) level and it was found to exist in two phases: a secretory phase (light cells) and a storage phase (dark cells). In the present investigation,H. fossilis was subjected to artificial photoperiods of continuous illumination and continuous darkness for a period of ten days and the effect on the secretory pinealocytes was studied at the EM level. Marked results were observed within the short period of ten days emphasizing the role of environmental photoperiod on the secretory activity of the pinealocytes. During continuous illuminated phase, both light and dark cells were observed: the light cells showed intense secretory activity and dark cells a storage one. During the dark phase both types of cells were present but in different metabolic states and neither of the cells demonstrated synthetic nor storage activity. Light cells were metabolically active but not secretory active and dark cells showed a necrotic condition. Phagocytotic activity of the dark cells was also seen. Intense neural activity was also observed during exposure to both the artificial photoperiods. The results highlight the role of light on the secretory activities of the pinealocytes of the catfish pineal organ.     

Neuronal organization of the medial part of the ventral horn of the cat spinal cord

An electron-microscopic study was made of the normal structure of the medial part of the ventral horn (Rexed's laminae VII and VIII) in the cervical portion of the cat's spinal cord, the region where fibers of reticulospinal and vestibulospinal tracts terminate. Neurons of this region can be divided on the basis of the density of their cytoplasmic matrix into "light" and "dark," the dark being much more numerous in this area (26% of the total number counted) than in other parts of the gray matter of the spinal cord. The mean diameter of the soma of the dark cells is smaller than that of the light cells, and it usually is 15–20 µ. Dendrites of the neurons can also be subdivided into "light" and "dark" respectively. The surface of the former is comparatively simple in shape with a small number of appendages and spine-like structures. On the surface of the dark dendrites there are many projections and irregularly shaped lacunae. The glial cells and their processes often completely cover the surface of the soma of the small neurons, and synaptic endings are found on it only where the dendrites leave the soma. Analysis of 1000 randomly chosen synaptic endings showed that 76.1% of them form axo-dendritic synapses, 14.2% axo-somatic, and 9.7% axo-axonal synapses. Of the total number of endings 50.9% contain spherical and 40.9% flattened synaptic vesicles. Some synaptic endings contain special structures under the postsynaptic membrane and have osmiophilic synaptic vesicles. The possible functional role of the pattern of neuronal organization revealed in this region is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 4, No. 2, pp. 176–183, March–April, 1972.     

Stratification and synaptogenesis in the mushroom body of the honeybee,Apis mellifera

Stratification is a basic anatomical feature of central brain in both vertebrates and many invertebrates. The aim of this study was to investigate the relationship between stratification and synaptogenesis in the developing mushroom bodies of the honeybee. During metamorphosis, the vertical lobe of mushroom body shows progressive stratification with three thick primary strata and more secondary strata and laminae. Three primary strata are formed at the metamorphic stage P1, before the youngest generation of the mushroom body intrinsic neurons, Kenyon cells, is produced. Thus, the primary strata within the lobe are unlikely to represent three major subpopulations of the Kenyon cells sequentially produced in the mushroom bodies. Formation of laminae starts at the stage P2 and culminates at the end of metamorphosis. The laminae appear within the lobe rather than being added sequentially from the ingrowth stratum. Alternating dark and light lamina (lamina doublets) are formed in the vertical lobe in late metamorphosis (stages P6–P9), but they are not visible in adults. The pattern of stratification is not continuous along the vertical lobe at the same developmental stage, and resorting of axons of the Kenyon cells is likely to occur within dark laminae. In the developing vertical lobe, dark laminae show lower synaptic density and exhibit an ultra structure that is indicative for a delay in synaptogenesis relative to the primary strata. A local transient block of synaptogenesis within the dark laminae may provide correct targeting of Kenyon cells by extrinsic mushroom body neurons. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Proctolin-immunoreactive neurons persist during metamorphosis of an insect: A developmental study of the ventral nerve cord of Tenebrio molitor(Coleoptera)

Summary Proctolin-immunoreactive neurons in all neuromers of the ventral nerve cord of Tenebrio molitor L. have been quantitatively demonstrated and mapped throughout metamorphosis. Each neuromer contains an anterior and a posterior group of neurons with light and dark staining properties as revealed by peroxidase-antiperoxidase labeling. Serial homologous subsets of dark staining neurons with central and peripheral projections have been identified and found to persist during morphogenetic changes from the larva to the adult. Most neurons maintain their topological and structural characteristics throughout metamorphosis. The identified proctolin-immunoreactive neurons exhibit structures similar to those described in other insect species; some may correspond known motoneurons.     

A possible mechanism of dopamine-induced synergistic disinhibition of thalamic cells via "direct" and "indirect" pathways in basal ganglia

On the basis of the mechanism of synaptic plasticity that we have earlier suggested for striatal spiny neurons and with regard to the known data about the predominance of dopamine-sensitive D1/D2 receptors on the striatonigral/striatopallidal cells it is hypothesized that the induction of the long-term potentiation/depression of the efficacy of excitatory cortical inputs to these cells can underlie the excitatory/inhibitory effect of dopamine on the activity of neurons that originate the "direct"/"indirect" pathways through the basal ganglia. Both these effects will lead to an enhancement of the activity of thalamic cells and activity of the efferent neocortical neurons excited by thalamic cells. The long-term potentiation of corticostriatal inputs to striosomal neurons, where, predominantly, D1 receptors are located, can also be induced by dopamine. This effect can be responsible of a rise of inhibition of dopaminergic cells and decrease in dopamine release by these cells. Such an event sequence can provide a stable dopamine concentration in the loop neocortex-basal ganglia-thalamus-neocortex.     

The influence of fixation on the relative amount of cytoplasmic ribosomes in mouse epidermal basal keratinocytes. A morphometric study of so-called "dark cells" and their putative role in epidermal carcinogenesis

The nature and significance of so-called dark keratinocytes in the epidermis during chemical carcinogenesis is still a matter of concern and debate. Based on ultrastructural observations it has been suggested that dark cells most often are shrunken cells. Reports on skin carcinogenesis, however, claim that dark cells are a sign of ongoing tumor promotion and represent those stem cells in the epidermis from which the tumors originate. It is therefore important to find out whether these cells are simply injured and shrunken cells, or vital cells of great importance for carcinogenesis. Dark cells are assumed to be rich in ribosomes. There is evidence, however, that the observed number of dark cells is highly dependent on tissue fixation. In the present ultrastructural study, morphometric methods were used to compare the effects of two different fixation procedures on the amount of cytoplasmic ribosomes in dark cells from both untreated and carcinogen-treated hairless mouse epidermis. The results show that the ultrastructural features of both dark and clear cells vary considerably with different fixation procedures. In acetone-treated controls typical dark cells are only observed when the fixative has a lower osmotic activity than the plasma. With iso-osmolal fixation typical dark cells are not observed. After an abortive two-stage carcinogenesis treatment, in which a single application of 9,10-dimethyl-1,2-benzanthracene (DMBA) in acetone was followed by a single application of 12-O-tetradecanoyl-13-acetate (TPA) in acetone, signs of cell injury could be found after both fixation procedures. With DMBA/TPA and hypo-osmolal fixation the number of dark cells seemed to increase, whereas only signs of cell injury with occurrence of some heavily altered "clear cells" dominated the picture with iso-osmolal fixation. Morphometry showed that both the numerical and the volumetric densities of cytoplasmic ribosomes in basal keratinocytes varied most significantly with the fixation procedure used. The cytoplasmic volumes did not vary in a way that could explain these differences. One might therefore assume that the number of ribosomes depends on the fixative. Large swelling artifacts occurred when a fixative with low osmotic activity was used, leading to compression of neighboring cells. Hence, an increased ribosomal density reported previously in dark cells is probably related to such cell volume artifacts and does not reflect an actually increased quantity of ribosomes.(ABSTRACT TRUNCATED AT 400 WORDS)     

The influence of fixation on the morphology of mouse epidermis. A light and electron microscopical study with special reference to "dark cells" and epidermal carcinogenesis

Different opinions exist on the normal ultrastructure of the epidermis including the significance of so-called basal dark cells. Thus, the dark cells are still assumed to be key elements in experimental skin carcinogenesis. We therefore explored the effects of tissue fixation on the ultrastructure of the epidermis. Untreated normal hairless mouse skin was processed for transmission electron microscopy with two different sets of fixatives, applied either by perfusion-immersion or immersion fixation only. The morphology of both the basal and the lower suprabasal layers of the epidermis, including the extracellular space, the shape and volume of the cells, their electron density, and the organisation of some of the organelles, were profoundly affected by the choice of fixatives. The non-keratinocytes showed comparable changes, including the appearance of a dark phenotype. The incidence of small electron-dense keratinocytes (dark cells) and the nature of their ultrastructure changed markedly with the fixation procedure. We were not able to identify undifferentiated dark cells. The pattern of changes and the quality of the morphological picture were almost unaffected by the mode of fixation. The upper suprabasal and the cornified layers appeared to be more or less unaltered by the change in fixatives and the method of application. The vehicle osmolality of the primary fixative was found to be mainly responsible for the ultrastructural appearances. A low vehicle osmolality may be responsible for the occurrence of the dark cell phenomenon, by inducing swelling artefacts of many cells with compression of some neighbouring cells.     

The filum terminale of the frog spinal cord

Summary The filum terminale, or terminal portion of the spinal cord, was studied in normal adult frogs (Rana pipiens) by means of light and electron microscopy. Astroglial cells are the predominant elements in this region. The rostral portion of the filum terminale consists mainly of (1) a peripheral dense ring of myelinated and some unmyelinated nerve fibers, and processes of astrocytes terminating at the subpial space; (2) an intermediate zone, in which astrocytes are the main cellular elements in addition to a few degenerated neurons; and (3) a central region where the central canal is lined by dark and light ependymal cells. In the caudal portion of the filum terminale, the amount of neuropil is greatly reduced. This region is formed mainly by astrocytic glial cells and very few neuronal elements. The central canal in the caudal portion is located ventrally and contains a lining consisting almost exclusively of dark ependymal cells.     

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     

Cholinergic neuronotrophic factors: VI. Age-dependent requirements by chick embryo ciliary ganglionic neurons

During development, ciliary ganglionic neurons become postmitotic and extend neurites in apparent independence of the presence of their future intraocular innervation targets. After reaching their peripheral innervation territory, however, these neurons become target dependent and about half of them die. We have previously reported that chick embryo intraocular target tissues contain a ciliary neuronotrophic factor (CNTF), which can be extracted and partially purified in a soluble form and which ensures near-total survival of 8-day chick embryo ciliary ganglionic neurons in monolayer cultures. In this study we have dissociated and cultured ciliary ganglia from embryonic Day (ED) 5 through 14, and examined dependence and responsiveness of their neurons to exogenously added CNTF. Two cell classes (dark and bright) could be distinguished by phase microscopy and differentially counted in cell dissociates from ED7–14, but not in ED5–6 ones. Dark cell number per ganglion increased from 6000 to 78,000 over this developmental time period. In contrast, bright cells (putative neurons) declined from a maximum of about 10,000 to 6000, suggesting a correlation with the expected neuronal cell death in vivo. Dissociated cells from ED5–14 ganglia were seeded on a polyornithine substratum coated with neurite promoting factor, cultured for 24 hr with or without added CNTF, and numerically examined for survival and neuritic development. Cultures from ED7–14 ganglia showed two cell categories: (i) flat nonneuronal elements dramatically increased in number with ganglionic age (thereby correlating with the increasing number of dark cells in the dissociates) and (ii) large, bright cells (often displaying neurite outgrowth) decreased in number in parallel with bright cell number in the dissociate. The survival of these neuronal elements was strictly dependent on exogenously added CNTF between ED7 and 10, but became progressively independent with older ages. ED14 neurons (fully capable of surviving for 24 hr without added CNTF) continued to require CNTF for neurite extension, thus displaying retained sensitivity to this factor. Although the ED5–6 cultures contained well-recognizable flat cells, the dominant category comprised cells with variable morphology, practically all of which exhibited neurite-like processes. Both the survival and neurite extension of these cells, which we tentatively interpret as immature neurons were independent of the presence of added CNTF.     

Are there efferent synapses in fish taste buds?

In fish, nerve fibers of taste buds are organized within the bud's nerve fiber plexus. It is located between the sensory epithelium consisting of light and dark elongated cells and the basal cells. It comprises the basal parts and processes of light and dark cells that intermingle with nerve fibers, which are the dendritic endings of the taste sensory neurons belonging to the cranial nerves VII, IX or X. Most of the synapses at the plexus are afferent; they have synaptic vesicles on the light (or dark) cells side, which is presynaptic. In contrast, the presumed efferent synapses may be rich in synaptic vesicles on the nerve fibers (presynaptic) side, whereas the cells (postsynaptic) side may contain a subsynaptic cistern; a flat compartment of the smooth endoplasmic reticulum. This structure is regarded as a prerequisite of a typical efferent synapse, as occurring in cochlear and vestibular hair cells. In fish taste buds, efferent synapses are rare and were found only in a few species that belong to different taxa. The significance of efferent synapses in fish taste buds is not well understood, because efferent connections between the gustatory nuclei of the medulla with taste buds are not yet proved.