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Sigmund Freud's contribution to the history of the neuronal cytoskeleton.

In the summers of 1879 and 1881, while Freud was a research student in Ernst von Brücke's laboratory at the Institute of Physiology at the University of Vienna, he carried out an important though seldom remembered investigation on the internal structure of nerve fibers and cells. His contribution to this field is here examined in the context of the 19th-century debate regarding the existence of neurofibrils and of present views on the cytoskeleton. Freud was able to discern separate fine fibrils following straight courses within the nerve fibers, as well as concentric loops of striae surrounding the nuclei and converging towards the processes of the cell bodies in crayfish nervous tissue. He thus confirmed and extended observations made by Robert Remak almost 40 years earlier, which had remained controversial. Electron microscopy of the crustacean nervous system confirmed Freud's main points, which in turn vindicated those of Remak. Both researchers were looking at small bundles of microtubules, and thus they were among the first to picture the lacy intracellular framework that future cell biologists would call the cytoskeleton.

Austria↗

Rapid Bielschowsky silver impregnation method using microwave heating.

The Bielschowsky silver impregnation method has been used extensively to demonstrate neuronal processes including dendrites, axons and neurofibrils. In this study, we examined the differences in the time required for and the staining quality of the Bielschowsky method for neuronal processes when microwave heating was used instead of processing at room temperature. For this purpose, a control group of sections stained according to the conventional method at room temperature was compared to an experimental group stained in a microwave oven at 180 W for 2, 4 and 1 min in 2% silver nitrate, ammoniacal silver nitrate and gold chloride, respectively. Light microscopic examination demonstrated that the normal structure was preserved in both groups and that there was no difference in the staining quality between the control and the microwave groups. In addition, staining time for this procedure was reduced to 8 min by using the microwave oven. Our study revealed that microwave irradiation can be used safely for Bielschowsky silver impregnation of neuronal tissues.

Animals↗

The Nissl substance of living and fixed spinal ganglion cells. I. A phase contrast study.

Living chick embryo spinal ganglion neurons grown from 1 to 4 weeks in vitro were studied under the phase contrast microscope. In the peripheral cytoplasm of the earliest stages studied, a homogeneous, phase-dense material is seen which corresponds in location to the cytoplasmic basophil material of the same stages. As maturation proceeds, this material increases in extent, and becomes separated by lighter channels into discrete bodies. Short fixation by 1 per cent buffered osmium tetroxide followed by post-fixation with neutral buffered formalin does not significantly alter the size, shape, or distribution of any of the cytoplasmic components, and the fixed, hydrated cell is almost indistinguishable from the living cell. Dehydration causes some shrinkage of the fixed preparations, but if the photographs of the stained preparations are enlarged to correspond with those of the living cell, excellent correspondence can be made between at least the larger basophil masses and the larger dark masses seen with phase contrast. Fixation by a formalin-mercuric chloride procedure also results in satisfactory correspondence between the stained Nissl bodies and the phase-dark homogeneous areas. It is concluded that discrete Nissl bodies preexist in the living neuron and are essentially unchanged after good cytological fixation. Evidence is also presented of the presence of neurofibrils in the living state.

Animals↗

Some observations on the fine structure of the sinus gland of a land crab, Gecarcinus lateralis.

The dilated axon endings of the sinus glands of the brachyuran crab, Gecarcinus lateralis, are filled with homogeneously dense granules, each granule being bounded by a delicate membrane. The granules are of two orders of magnitude: 0.05 to 0.1 micro and 0.15 to 0.2 micro in diameter. Each axon ending contains granules of a nearly uniform size. Endings with granules of the larger size range predominate. Non-nervous cells endogenous to the sinus gland are scattered among the nerve endings. The cell contours are irregular, and cytoplasmic processes ramify between endings. The axons are unmyelinated, having only thin limiting membranes, and they possess many neurofibrils. Granules in preterminal portions of the axons tend to lie at the periphery of the fiber, and in some cases in chains at the core of the fiber. The granules appear to be storage and release centers for neurosecretory substances or their precursors.

Animals↗

Electron microscopic study of demyelination in an experimentally induced lesion in adult cat spinal cord.

Plaques of subpial demyelination were induced in adult cat spinal cords by repeated withdrawal and reinjection of cerebrospinal fluid. Peripheral cord was fixed by replacing cerebrospinal fluid available at cisternal puncture with 3 per cent buffered OsO(4). Following extirpation, surface tissue was further fixed in 2 per cent buffered OsO(4), dehydrated in ethanol, and embedded in araldite. Normal subpial cord consists mainly of myelinated axons and two types of macroglia, fibrous astrocytes and oligodendrocytes. Twenty-nine hours after lesion induction most myelin sheaths are deteriorating and typical macroglia are no longer visible. Phagocytosis of myelin debris has begun. In 3-day lesions, axons are intact and their mitochondria and neurofibrils appear normal despite continued myelin breakdown. All axons are completely demyelinated by 6 days. They lack investments only briefly, however, for at 10 and 14 days, macroglial processes appear and embrace them. These macroglia do not resemble either one of the normally occurring glia; their dense cytoplasm contains fibrils in addition to the usual organelles. It is proposed that these macroglia, which later accomplish remyelination, are the hypertrophic or swollen astrocytes of classical neuropathology. The suggestion that these astrocytes possess the potential to remyelinate axons in addition to their known ability to form cicatrix raises the possibility of pharmacological control of their expression.

Animals↗

Cell-surface ganglioside GD2 in the immunohistochemical detection and differential diagnosis of neuroblastoma.

The expression of the disialoganglioside GD2 was analyzed in 67 solid tumors and normal tissues from children by using the GD2-specific murine monoclonal antibody 3A7 and the indirect immunoperoxidase method. GD2 was expressed in all of 28 neuroblastomas and was most abundant in stroma-poor tumors. In differentiating stroma-rich neuroblastomas, neuroblastic clusters, neurofibrils, and most ganglion-like cells were positive, whereas Schwann's-cell stroma did not express GD2. In ganglioneuromas, only a few ganglion-like cells showed GD2, whereas all other structures were negative. Scattered foci of ganglioside GD2 also were found in some non-neuronal tumors, such as rhabdomyosarcomas and osteosarcomas, but not in lymphomas, Askin tumors, or most Wilms' tumors. The monoclonal antibody 3A7 is a useful aid in the immunohistochemical diagnosis of neuroblastoma. In addition, the intense cell surface staining of neuroblastoma cells by this reagent makes it potentially useful for detecting residual neuroblastoma in bone marrow samples and lymph node biopsies.

Antibodies, Monoclonal↗

Localization and characterization of calcitonin gene-related peptide mRNA in rat heart.

Calcitonin gene-related peptide (CGRP), a product of the calcitonin/CGRP gene, is a potent vasodilating neuropeptide widely distributed throughout the cardiovascular system, particularly in the heart. Immunocytochemical studies have demonstrated CGRP-containing neurofibrils in the myocardium and in the periadventitia of coronary blood vessels. Based on these studies, it has been assumed that all of the CGRP peptide in the heart is synthesized in neurons whose cell bodies are located outside of the heart. Using Northern blot analysis and a ribonuclease protection assay, we observed in the rat heart low levels of a CGRP-like mRNA species that appeared to be identical to authentic CGRP mRNA produced in the brain and dorsal root ganglia. The ventricles contained somewhat more CGRP mRNA than did the atria. Also, whereas the dorsal root ganglia synthesized both alpha- and beta-CGRP mRNA, only the alpha-CGRP mRNA was detected in the heart. The presence of CGRP mRNA in the heart suggests that the CGRP gene is transcriptionally active in a subpopulation of heart cells, possibly neuronal, which have the potential to synthesize and secrete this neuropeptide. Given the potent coronary vasodilatory and positive chronotropic and inotropic effects of CGRP, the localized synthesis of CGRP in the heart may play a role in modulating cardiovascular function.

Animals↗

Identification of "premyelination" by diffusion-weighted MRI.

OBJECTIVE: The purpose of this study was to compare white matter maturation as demonstrated with diffusion-weighted MRI and with myelin-sensitive histological staining. MATERIALS AND METHODS: The diffusion-, T1-, and T2-weighted SE MRI at 4.7 T was performed weekly in a total of 16 rat pups, aged from 5 days to 8 weeks, 2 animals evaluated per week. Heavily diffusion-weighted sequences were obtained with the diffusion-sensitizing gradient switched alternately in two orthogonal directions. To enhance signal intensity of anisotropic structures, a synthesized image (referred to as the "anisotropy index map") was constructed from the ratio of pairs of images acquired with diffusion sensitization of identical magnitude but orthogonal direction sensitivity. The anisotropy index maps were used for comparison with T1-weighted and heavily T2-weighted SE sequences and histological sections, respectively. RESULTS: The first evidence of diffusion anisotropy on anisotropy index maps preceded initial myelin as well as neurofibril staining by 5-12 days and T2 shortening by 2 weeks. The T1-weighted sequences did not yield visible changes and were not helpful for the assessment of ongoing white matter maturation in this model. CONCLUSION: Magnetic resonance imaging signal intensity changes based on anisotropic water diffusion were demonstrated in regions of unmyelinated cerebral white matter tracts of albino rat pups before the onset of histologically detectable myelin. The ability of in vivo mapping of premyelinating white matter maturation indicates a new diagnostic use of MRI in evaluating cerebral white matter maturation.

Animals↗

Non-epithelial cellular components in eccrine spiradenoma: a histological and immunohistochemical study of 20 cases.

In this study, 20 cases of eccrine spiradenoma have been examined using monoclonal antibodies to identify the nature of the epithelial, as well as the non-epithelial, cellular components of this tumour. The results indicate that there is striking similarity between the epithelial cells of eccrine spiradenoma and the normal cells lining eccrine apparatus. An interesting finding was the presence of abundant T-lymphocytes and Langerhans' cells within the tumour lobules. Additionally, endothelial-lined vascular channels and neurofibrils were prominent in larger lesions.

Adenoma, Sweat Gland↗

Fine structural comparison of Ewing's sarcoma with neuroblastoma.

Two cases of Ewing's sarcoma and two neuroblastoma, rosette forming and round cell type, were studied electron microscopically and their fine structures were compared. The neoplastic cells of Ewing's sarcoma were characterized by aggregated glycogen particles in the cytoplasm. They had pseudopod-like cytoplasmic processes having tight junctions, which never contained microtubules or mitochondria. Ewing's sarcoma cells exhibited several stages of cell maturation and some mature cells possessed a large amount of smooth and rough endoplasmic reticulum, large Golgi complexes and numerous phagosomes containing glycogen particles as well as cytoplasmic organelles. The neoplastic cells of neuroblastoma, rosette forming type, were characterized by synaptic junctions and numerous cytoplasmic processes with production of neurites containing microtubules, neurofibrils, mitochondria and a few catecholamine granules. A few cytoplasmic processes containing mitochondria were observed even in the round cell type.

Adolescent↗

Glucocorticoid exposure at the dose used clinically alters cytoskeletal proteins and presynaptic terminals in the fetal baboon brain.

Glucocorticoids have been used for 30 years to accelerate fetal lung maturation in human pregnancy at risk of preterm delivery. Exposure to inappropriate levels of steroid, however, leads to altered maturation of the cardiovascular, metabolic and central nervous systems. The effects of betamethasone on neuronal development and function were determined in the fetal baboon brain by examination of cytoskeletal microtubule associated proteins (MAPs) and the presynaptic marker protein synaptophysin. At 0.73 gestation, commencing 28 weeks of gestation, pregnant baboons received four doses of saline (n = 8) or 87.5 microg (kg body weight)(-1) betamethasone I.M. (n = 7) 12 h apart. This dose is equivalent to 12 mg betamethasone administered daily over two consecutive days to a 70 kg woman. Baboons underwent Caesarean section 12 h after the last injection. Paraffin sections of the fetal neocortex and the underlying white matter were labelled immunohistochemically against MAP1B, MAP2abc, MAP2ab and synaptophysin and stained histochemically with hematoxylin-eosin and silver. Tissue staining was quantified morphometrically. Betamethasone exposure resulted in decreased immunoreactivity (IR) of MAP1B by 34.3 % and MAP2abc by 34.1 % (P < 0.05). Loss of MAP2 IR was due to loss of IR of the juvenile isoform MAP2c (P < 0.05). MAP1B and MAP2c are involved in neuritogenesis and neuronal plasticity. Synaptophysin IR was reduced by 51.8 % (P < 0.01). These changes might reflect functional neuronal disturbances because they were not accompanied by an alteration of the density of neurofibrils or neuronal necrosis. These results are in agreement with earlier findings of alterations of cytoskeletal proteins and presynaptic terminals in the fetal sheep brain after betamethasone infusion directly to the fetus and support a common effect of inappropriate fetal exposure to glucocorticoids on neuronal cytoskeleton and synapses in mammalian species.

Animals↗

Neuronal cells from chick embryo cerebral hemispheres cultivated on polylysine-coated surfaces.

Dissociated cells from 5- to 12-day-old chick embryo cerebral hemispheres were cultivated in polylysine-coated plastic Petri dishes. The polylysine substrate was observed to be favorable for the growth of neuronal cells, whereas glioblast proliferation was inhibited. The optimal conditions for the production of a predominantly neuronal culture were to use cerebral hemispheres from 7-day-old chick embryos, to dissociate the brain tissue mechanically and to seed the cells at a concentration range between 1.5 and 5 X 10(6) cells/ml. The cultures were observed by phase contrast microscopy. Most cells grew fibers and differentiated into bipolar and multipolar neurons. These neurons were stained by thionine, which demonstrated the presence of Nissl bodies. The silver impregnation revealed the presence of neurofibrils within the nerve fibers. Acetylcholinesterase was found to be present in the neuronal cells, but absent in the glioblasts. Under our culture conditions the neurons survived for 10-12 days. This system should allow further studies on the effects of growth factors on the differentiation of isolated neurons as well as investigations on neuron-glial interrelationship.

Animals↗

Morphogenesis of cerebellar lesions in bovine familial convulsions and ataxia.

Cerebellar lesions develop in a heritable disorder characterized by recurrent episodic seizures in newborn and young calves and by ataxia in survivors of several bouts of convulsions. Earliest changes were altered patterns of phosphatase reaction products in Purkinje cells. Purkinje cells axons in the outer half of the granular layer developed fusiform or spheroidal argyrophilic swellings. Early lesions were restricted to the lingula and uvula, but cases with more persistent clinical disease involved other parts of the vermis. In ataxic calves these lesions were also in the cerebellar hemispheres. The axonal swellings showed proliferation of tubulovesicular endoplasmic reticulum, neurofibrils and mitochondria. In other swellings axoplasm was degenerate, and in some it resembled Purkinje cell cytoplasm. In some clinically normal adult transmitters of the disorder, light microscopy showed some similar swellings, but most calves that recovered had no lesions.

Animals↗

Paragangliomas of the head and neck: ultrastructural and immunohistochemical analysis.

Eighteen head and neck paragangliomas were studied by light microscopy and light microscopic immunohistochemistry by the peroxidase technique for the presence of NSE (neuron-specific enolase), serotonin, and a battery of neuropeptides. Seven of these tumors were also studied by electron microscopy. All 18 cases demonstrated immunostaining for NSE; 10 of the 11 carotid body tumors had immunostaining for multiple hormones. Considering all 18 cases, the most frequently demonstrated hormonal substances were in order: serotonin, leu-enkephalin, gastrin, substance P, vasoactive intestinal polypeptide (VIP), somatostatin, bombesin, calcitonin, and alpha MSH. In several tumors, adjacent-step sections stained for different hormonal substances strongly suggested reactivity for more than one hormone in given tumor cells. By electron microscopy, all 7 cases studied displayed considerable heterogeneity of the neurosecretory granules with respect to size, shape, and electron density. This demonstrated that branchiomeric paragangliomas are capable of producing a spectrum of neuropeptides in addition to their known amine content. The presence of immunoreactive serotonin in most of these neoplasms was confirmed. In addition to these findings, neurofibrils within the substance of carotid body paragangliomas demonstrated immunoreactivity for somatostatin and a gastrinlike neuropeptide. The significance of the neuropeptides in these neoplasms and their possible presence and role in normal and hyperplastic paraganglia remain to be defined.

Carotid Body Tumor↗

A combination staining method for fibers and cell bodies of the urodele central nervous system.

A simple method for staining nerve cells and fibers of the salamander central nervous system is described. The procedure employs Carnoy's fixation followed by Protargol inpregnation and Nissl staining. This technique permits the simultaneous observation of intracellular neurofibrils, neuronal processes and basophilic components of the neuron. In addition, it eliminates the need to stain alternate sections with separate procedures to view the various components of the urodele central nervous system.

Animals↗

[Therapy of Alzheimer dementia. Current status and prospects].

In recent years, considerable progress has been made both in the diagnosis and treatment of dementia. Drugs have been developed which enhance cognitive performance in a large percentage of those afflicted, delay impairment of the ability to cope with the tasks of daily life, and avoid premature admission to a nursing home. In the practical medical care setting, however, these advantages are being utilized to only a limited extent, and this despite the fact that numerous therapeutic options are now available for the treatment of AD. New therapeutic approaches are aimed at inhibiting the pathological cleavage and extracellular and intracellular deposition of amyloid, preventing the toxic effects of amyloid accumulation around the neurons, and re-establishing intracellular transport deranged by the aggregation of neurofibrils. A further approach is the use of combinations of available substances with differing, possibly synergic, effects. Over and beyond this, treatment of AD in the "presymptomatic stage", or in the stage of only mild cognitive disturbance, is currently the subject of clinical trials.

Aged↗

[Age changes in rat vasomotor reflexes and sympathetic neuron ultrastructure following chemical sympathectomy].

Prolonged administration of guanethidine (20 mg/kg) to newborn rats caused a marked reduction in the number of cells in stellate ganglia. The administration of guanethidine for 14 days decreased the amount of cells to 30% of the normal (partial sympathectomy), and for 28 days--to 0.5% (complete sympathectomy). At the age of two months the blood pressure pressor reflexes to asphyxia and femoral nerve stimulation were absent in both groups of the sympathectomized animals. These responses, however, were restored in the partially sympathectomized animals at the age of four months. No restoration took place in the completely sympathectomized animals. The electron microscopic studies of neurons in the partially sympathectomized animals showed the presence of a great number of neurofibrils. According to literature data this fact was typical of cells in which an active growth of axon fibers took place.

Aging↗

[Esthesioblastoma of the nose].

Morphological investigations of biopsy materials in 8 cases of esthesioblastomas of the nasal cavity were carried out. The three morphological variants of the tumour are described: esthesioneuroepithelioma, esthesioneurocytoma, and esthesioneuroblastoma. Rare occurrence of esthesioblastomas, peculiar features of their histological structure and difficulties of diagnosis in histological verification are emphasized. Lymphosarcoma is recommended to be differentiated from nondifferentiated cancer and sympathoblastoma. An important differentially-diagnostic procedure is considered to be a positive reaction to argentation (for identifying neurofibrils).

Adolescent↗