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PubMed1996

Treatment of oligodendrocytes with antisense deoxyoligonucleotide directed against CREB mRNA: effect on the cyclic AMP-dependent induction of myelin basic protein expression.

We have shown previously that in oligodendrocytes, the transcription factor cyclic AMP response element binding protein (CREB) is maximally expressed immediately prior to the most rapid period of myelination in rat brain. We have begun to investigate the role of this protein during myelination by downregulating CREB synthesis in cultured oligodendrocytes using an antisense deoxyoligonucleotide directed against CREB mRNA. Neonatal oligodendrocytes were grown for 4 days in a chemically defined medium (CDM) after which intracellular delivery of CREB antisense oligonucleotide was facilitated by using a liposome preparation. Control cultures were treated in a similar manner but in the presence of CREB sense oligomer. Immediately after transfection, cells were cultured for 3 days in CDM in the presence or absence of the cyclic AMP (cAMP) analogue N6, O21-dibutyryl cAMP (db-cAMP). In these cultures, myelin basic protein (MBP) expression was investigated by immunocytochemistry and Western blot analysis. Treatment of control cultures with db-cAMP resulted in a significant increase in the number of MBP positive cells which was abolished when the cells were treated with CREB antisense oligonucleotide. MBP positive cells in control cultures treated with db-cAMP have extended and highly branched MBP positive processes. In contrast, MBP positive cells in either control cultures grown in the absence of db-cAMP or cultures grown in the presence of db-cAMP but treated with CREB antisense oligonucleotide showed shorter and less complex processes and the MBP immunoreactivity appeared to be concentrated in the cell body. These observations suggest that CREB is at least one of the mediators in the induction of oligodendrocyte differentiation by cAMP.

Activating Transcription Factor 2↗
PubMed2001

Nidus and tasseled cell: distinctive neuronal organization of the main olfactory bulb of the laboratory musk shrew (Suncus murinus).

We revealed the structural features of particular synaptic regions, nidi, and newly found neurons, tasseled cells, in the main olfactory bulb (MOB) of the laboratory musk shrew (Suncus murinus). Nidi were intensely immunoreactive for glutamic acid decarboxylase (GAD) and calbindin D28k (CB), were 30-80 microm in diameter, and were located beneath glomeruli, appearing to make glomerulus-nidus unit-like complexes. In contrast to glomeruli, they contained few or no olfactory nerves. Nidi were distributed throughout the whole MOB and made a distinctive layer, nidal layer. Tasseled cells were located in the mitral cell layer and in the middle of the external plexiform layer (EPL) and extended single primary dendrites to the nidus, where their small tuft-like complicated branches intermingled with processes of perinidal cells surrounding nidi. Primary dendrites of mitral/tufted cells also penetrated nidi but passed to glomeruli. In the outer half of the EPL, columnar structures were seen, where CB- and GAD-positive elements appeared to associate with bundles of cylindrical dendrites of presumed mitral/tufted and tasseled cells. By electron microscopic examinations, nidi were confirmed to be particular synaptic areas where GAD-positive processes made symmetrical synapses to GAD-negative presumed tasseled and mitral/tufted cell dendrites and received asymmetrical synapses from the latter. Retrograde tracings revealed that tasseled cells, in addition to mitral/tufted cells, projected their axons to the lateral olfactory tract, indicating that there were two parallel projection systems in the shrew MOB, which might interact with each other via various types of gamma-aminobutyric acid (GABA)ergic interneurons. The present study clearly showed that the neuronal organization of the shrew MOB was distinctly different from that in rodents.

Animals↗
PubMed1987

Microtubules, intermediate filaments, and actin filaments in the odontoblast of rat incisor.

Actin filaments, intermediate filaments, and microtubules in the odontoblasts of rat incisors were investigated electron microscopically using heavy meromyosin and taxol. Actin filaments were abundant at the periphery of the odontoblast process in the form of a network or in bundles. In a branch of the odontoblast process, longitudinally oriented actin filament bundles were found. Most actin filaments were associated with the plasma membrane via electron-dense material which stained with tannic acid. The intermediate filaments had a diameter of 11 to 13 nm. They were distributed throughout the cytoplasm of odontoblasts. They ran lengthwise in the core of the odontoblast process, which showed a different distribution compared with that of actin filaments. Microtubules, which were disrupted after Triton X-100 but preserved by addition of taxol, tended to be associated with intermediate filaments. Such a relation was also seen in conventional preparations. Coated vesicles, which were abundant at the periphery of the odontoblast process, were often associated with actin filaments. Therefore, it is suggested that actin filaments, in the odontoblast process at least, play a role associated with the coated vesicles at the periphery of the process, and may be involved in coated vesicle transport.

Actins↗
PubMed2004

Correlation of electrophysiological and morphological characteristics of enteric neurons in the mouse colon.

We report on the first correlative study of the electrophysiological properties, shapes, and projections of enteric neurons in the mouse. Neurons in the myenteric plexus of the mouse colon were impaled with microelectrodes containing biocytin, their passive and active electrophysiological properties determined, and their responses to activation of synaptic inputs investigated. Biocytin, injected into the neurons from which recordings were made, was converted to an optically dense product and used to determine the shapes of neurons. By electrophysiological properties, almost all neurons belonged to one of two classes, AH neurons or S neurons. AH neurons had a biphasic repolarization of the action potential, and slow afterhyperpolarizing potentials usually followed the action potentials. S neurons had monophasic repolarizations, no slow afterhyperpolarization, and fast excitatory postsynaptic potentials in response to fibre tract stimulation. By shape, neurons were divided into Dogiel type II (28/136 neurons) and uniaxonal neurons. Dogiel type II neurons had large, smooth-surfaced cell bodies and several long processes that supplied branches within myenteric ganglia. All Dogiel type II neurons had AH electrophysiology; conversely, most AH neurons had Dogiel type II morphology. The majority of uniaxonal neurons had lamellar dendrites, i.e., Dogiel type I morphology. They projected to the circular muscle (circular muscle motor neurons), to the longitudinal muscle (longitudinal muscle motor neurons), and to other myenteric ganglia (interneurons) and in some cases could not be traced to target cells. All S neurons were uniaxonal. A small proportion of uniaxonal neurons (3/70) had AH electrophysiology. Fast excitatory synaptic potentials were only recorded from uniaxonal neurons and were in most cases blocked by nicotinic receptor antagonists. A small component of fast excitatory transmission in some neurons was antagonized by the purine receptor antagonist PPADS. Slow excitatory postsynaptic potentials were observed in both AH and S neurons. Slow inhibitory postsynaptic potentials were recorded from S neurons. We conclude that the major classes of neurons are Dogiel type II neurons with AH electrophysiological properties and Dogiel type I neurons with S electrophysiological properties. The S/Dogiel type I neurons include circular muscle motor neurons, longitudinal muscle motor neurons, and interneurons.

Action Potentials↗
PubMed1977

The developing caudate nucleus in the euthyroid and hypothyroid rat.

The basal ganglia are presently implicated in learning, and thyroid deficiency induced neonatally is known to affect mentation. The effects of such a deficiency on the developing causate nucleus might be used to provide insight into structure and function of the normal subcortical brain, as well as possible influences of these extrapyramidal structures on mental retardation. Propylthiouracil was added to the diet of lactating rat dams and observations of the developing caudate nuclei of normal hypothyroid rats were made at 8, 14, 20, 30 and 42 days by using various tissue stains and Golgi-Cox preparations. Seven different types of neurons were distinguished in the caudate nucleus. Differences in the size of cell somata and the varying morphology of axons and dendrites were criteria used to make distinctions. Normally, the nucleus acquires cytoarchitectural complexity during the first three postnatal weeks. Within this period, neuron incidence increases in the caudate neuropil with age while the germinal matrix density decreases. Neuron accumulation reaches a plateau after the third week and cell migration is essentially complete at the end of the first postnatal month as shown by computer analysis of Nissl stained cell counts. Branching of cellular processes, attainment of receptor spines and complexity of the fiber network also appeared during this period. Retardation of structural development with thyroid hormone deficiency was shown by decreased numbers of neurons, inhibition of dendritic arborization, decreased numbers of dendritic spines and a reduced complexity of axonal plexuses. Thyroid deficiency delays cell migration during the first three weeks when compared to age-matched normal controls. The lack of thyroid hormone does not appear to influence the size of neuron somata, and the extent of related dendritic fields, nor does hypothyroidism affect a specific cell type population. Generalized disturbances of caudate nuclear morphological maturation are caused by the deficiency. An apparent compensatory process, including a spurt of neural growth and differentiation, takes place in the period between days 14 and 30 in the deficient animals and a seemingly "normal" caudate cytoarchitecture is seen after the third postnatal week. Quantitative data, however, show that this rapid "catch up" process is inadequate. The developmental imperfection of the caudate nucleus which persists might be a part of the underlying substrate for the mental retardation, disturbed motor performance and perceptual handicaps which are found in the human patient.

Animals↗
PubMed1991

Vasoactive intestinal polypeptide-containing cells in the rabbit retina: immunohistochemical localization and quantitative analysis.

Vasoactive intestinal polypeptide (VIP) possesses neuroactive properties in the nervous system. In this study we characterized VIP immunoreactive neurons in the rabbit retina to provide a basis for a better understanding of the role of this peptide in retinal functions and to further define the morphology of wide-field amacrine cells. VIP immunoreactivity was demonstrated in colchicine-treated retinas. Immunolabeling was observed in amacrine cells located in the proximal inner nuclear layer and, occasionally, in the ganglion cell layer and inner plexiform layer (IPL). Varicose fibers were distributed in laminae 1, 3, and 5 of the IPL. The distribution of VIP immunoreactive cells showed a peak of approximately 50 cells/mm2 in the visual streak and minimum values of approximately 12 cells/mm2 in the peripheral retina. The total number of VIP immunopositive neurons was estimated to be about 11,000. Cell body diameters in the visual streak (8-9 microns) were slightly smaller than those measured in the dorsal or in the ventral retina (9-10 microns). The distribution of nearest neighbor distances (approximately 109 microns in the visual streak and approximately 99 microns in the peripheral retina) showed that VIP immunoreactive neurons were nonrandomly spaced. Labeled neurons emitted one to three thick primary processes, arborizing in secondary processes and collaterals rich in varicosities; these processes often crossed among different IPL laminae. Arborization fields of individual cells overlapped extensively. In the dorsal retina, estimated areas of single arborization fields were larger and processes had lower branching frequency than in the visual streak and in the ventral retina. On the whole, VIP immunoreactive amacrine cells gave rise to a loose meshwork of fibers in the IPL. These characteristics indicate VIP is contained in a class of wide-field amacrine cells and is likely to be involved in widespread regulatory or modulatory functions rather than in the direct transmission of visual information through the retina.

Animals↗
PubMed2004

Segmented detachable structure of cochlear-implant electrodes for close-hugging engagement with the modiolus.

Surface and bulk modification techniques of polydimethylsiloxane (PDMS) polymers were used to develop a new intracochlear electrode that can closely hug the inner wall of scala tympani. Laser-induced surface grafting of poly (2-hydroxyethyl methacrylate) (PHEMA) and sequential method for preparation of interpenetrating polymer networks (IPNs) of PDMS/PHEMA were, respectively, used for surface and bulk modifications. The hydrogel content and water-uptake capability of the modified samples were optimized by Taguchi method for experimental design. The modified PDMS samples were examined by performing attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, water contact-angle measurement, and peel strength tests. The performances of actual-sized fabricated electrodes were assessed inside a transparent model of scala tympani, which was filled with saline. After a swelling process, the hydrophilic branch begins to elongate and detach from hydrophobic branch and pushes it toward the inner wall.

Cochlea↗
PubMed1984

Biology of gangliosides: neuritogenic and neuronotrophic properties.

Research on the biologic function of gangliosides has accelerated in recent years following discovery of their pronounced effects when administered exogenously to neurons in culture and in vivo. These effects are of two principal types: 1) neuronotrophic, concerned primarily with survival and maintenance of the neuron, and 2) neuritogenic, involving significant increase in the number, length, and/or branching of neuronal processes. Such neurite-promoting activity has been observed in primary cultures of neurons from brain and ganglia as well as transformed lines of neuronal origin. These phenomena may be related to the remarkable growth of aberrant secondary neurites, often accompanied by synaptogenesis, observed in the gangliosidoses. Several in vivo studies have shown exogenously administered gangliosides to aid nervous system repair in both the CNS and PNS, although it is not clear in some cases whether the observed effects should be attributed to neuronotrophic or neuritogenic effects (or both). This article attempts to briefly review the principal developments that have occurred in this area of ganglioside research over the past several years. It also presents for consideration some of the tentative hypotheses put forward concerning mechanism of action.

Animals↗
PubMed1993

Effects of the steroid hormone 20-hydroxyecdysone and prior sensory input on the survival and growth of moth central olfactory neurons in vitro.

Neurons in the developing (antennal) olfactory lobe of the moth Manduca sexta undergo a period of extensive process outgrowth and branching that coincides temporally with both a rising titer of the steroid hormone 20-hydroxyecdysone and the ingrowth of sensory axons from receptors in the antenna. To evaluate the contribution of these two influences to the morphological development of antennal-lobe neurons, we placed the neurons in cell culture. Antennal-lobe neurons were dissociated from normal and chronically unafferented lobes at different stages of development and were exposed to different doses of hormone. Six neuronal cell types with distinctive and stable morphologies appeared in cultures from all stages of pupal development. Morphological changes in these neuronal types were examined quantitatively by comparison of the total length and number of branches. We found that 20-hydroxyecdysone had little direct effect on the morphological development of antennal-lobe neurons, but brief exposure to sensory axons in vivo prior to dissociation significantly enhanced subsequent outgrowth in culture.

Animals↗
PubMed1980

Neuronal damage in chick and rat embryos following X-irradiation.

Exposure of rat and chick embryos to X-irradiation at the time of development of neurons at the telencephalic-diencephalic border results in prolonged damage to neurons in this area as measured by neuronal nuclear size. A dose of 100 rads to the seven-day-old chick embryo has about the same effect as 125 rads to the 15-day-old rat fetus. The nuclear volume of large, multipolar neurons in the chick paleostriatum primitivum and the rat lateral preoptic area are reduced from 10 to 15%. Larger doses of X-irradiation to the chick (150 and 200 rads) cause progressively greater reductions in nuclear size. The large neurons which were measured in the rat and chick are morphologically similar in the two species. Both contain cytoplasmic acetylcholinesterase and have several branched, spiny dendritic processes. The similarity of response of chick and rat neurons to X-irradiation diminishes the significance of maternal factors as the cause of the effects of fetal irradiation in these experiments.

Animals↗
PubMed1999

Dynein-dynactin function and sensory axon growth during Drosophila metamorphosis: A role for retrograde motors.

Mutations in the genes for components of the dynein-dynactin complex disrupt axon path finding and synaptogenesis during metamorphosis in the Drosophila central nervous system. In order to better understand the functions of this retrograde motor in nervous system assembly, we analyzed the path finding and arborization of sensory axons during metamorphosis in wild-type and mutant backgrounds. In wild-type specimens the sensory axons first reach the CNS 6-12 h after puparium formation and elaborate their terminal arborizations over the next 48 h. In Glued1 and Cytoplasmic dynein light chain mutants, proprioceptive and tactile axons arrive at the CNS on time but exhibit defects in terminal arborizations that increase in severity up to 48 h after puparium formation. The results show that axon growth occurs on schedule in these mutants but the final process of terminal branching, synaptogenesis, and stabilization of these sensory axons requires the dynein-dynactin complex. Since this complex functions as a retrograde motor, we suggest that a retrograde signal needs to be transported to the nucleus for the proper termination of some sensory neurons.

Animals↗
PubMed1997

A new model for discrete character evolution.

The paper provides an explicit justification for the principle that a uniform taxon should contribute only one datapoint in comparative analyses with discrete variables. The justification is that phylogenetic patterns in variables unincluded in the proposed test vitiate the assumption of independence, both at the level of species and at the level of branch segments. The consequence is that a uniform taxon cannot safely be counted as more than one datapoint. The arguments use a branching discrete Markov process in continuous time, with the new feature that the tested variables are only a subset of the evolving characters. This model is proposed as a useful criterion for measuring the merit of proposed tests, and illustrates the necessity for models in evaluating comparative methods.

Adaptation, Biological↗
PubMed1983

GABA or sodium-bromide-induced plasticity of neurites of mouse neuroblastoma cells in culture. A quantitative study.

Differentiated C1300 mouse neuroblastoma cells were treated with 10(-4)-10(6) M gamma-aminobutyric acid (GABA) and/or sodium bromide (NaBr) for 2 days and then fixed. Quantitative studies revealed an increase in the length and branching of the processes, as well as an increase in the number of cells when compared to the controls. It is suggested that the above changes contribute to the augmentation of specialized contacts between cells and processes as well as the further maturation of the primitive stages of synaptogenesis as discussed.

Animals↗
PubMed1993

Some horizontal cells of the bovine retina receive input synapses in the inner plexiform layer.

Bovine retinae were stained immunocytochemically with antibodies against the calcium-binding protein, calbindin. Horizontal cells in the outer plexiform layer were heavily labelled. The processes of most horizontal cells were confined to the level of the outer plexiform layer, and the tips of their dendrites were positioned as the lateral elements of the cone triads, viz. the usual mammalian arrangement. However, some of the horizontal cells had additional thick processes descending to branch within the inner plexiform layer, where they were postsynaptic at bipolar cell dyads and where they also received input from amacrine cells. No output synapses of horizontal cells were observed in the inner plexiform layer.

Animals↗
PubMed1993

Cycloheximide mimics effects of oestradiol that are linked to synaptic plasticity of hypothalamic neurons.

The synaptic connectivity of the rat arcuate nucleus, a hypothalamic area rich in oestradiol receptors, is rapidly affected by physiological modifications of hormonal levels. A rise of oestradiol in plasma elicits a coordinated neuronal-glial response that begins with a rapid fall in the number of small (< 10 nm) intramembrane particles and a rapid increase in the number of large (> 10 nm) intramembrane particles in neuronal membranes, followed by a modification in the branching of astrocytic processes and finally results in decreased number of axo-somatic synapses and increased glial wrapping of the neuronal somas. In the course of a series of studies aimed to test possible non-genomic effects of oestradiol on neuronal membranes we analyzed the effect of the systemic administration of the protein synthesis inhibitor cycloheximide on the ultrastructure of arcuate neurons and granule cells of the cerebellar cortex, an area of the brain with low levels of estrogen receptors. Cycloheximide resulted in a significant inhibition of protein synthesis in hypothalmus and cerebellum of ovariectomized rats. Under these circumstances, the number of small intramembrane particles was reduced in hypothalamic and cerebellar neuronal membranes while the number of large intramembrane particles showed a decrease in cerebellar membranes and a transient increase in arcuate neuronal somas. Furthermore, cycloheximide resulted in an increased glial wrapping of arcuate neuronal somas but not of cerebellar granule cells. The ensheathing of arcuate neurons by glial was associated with a 41% decrease in the number of axo-somatic synapses. These results indicate that the protein synthesis inhibitor cycloheximide may elicit the integrated neuronal-glial response that is associated with the hormonally induced remodelling of synaptic contacts on arcuate neurons.

Animals↗
PubMedDate not supplied

Electroencephalographic biofeedback methodology and the management of epilepsy.

Currently considerable research is being directed toward developing methodologies for controlling internal processes. An applied branch of the basic field of psychophysiology, known as biofeedback, has developed to fulfill clinical needs related to such control. Current scientific and popular literature abounds with numerous examples of how biofeedback is being used. For example, germinal studies by Kamiya (1962), and later work by Lynch and Paskewitz (1971), Beatty (1973), as well as many others have shown that the EEG alpha rhythm (8 to 13 Hz) recorded from occipital regions of the human brain can be behaviorally manipulated when feedback or reward is provided for changing the density of this activity. Other researchers have provided evidence that theta activity (4 to 7 Hz) and the beta activity (greater than 14 Hz) can also be controlled by humans and analogs of this activity have been conditioned in animals as well (Green, Green and Walters, 1971). In addition to the work that has been carried out with the EEG, researchers such as Engle and Bleecker (1973) have indicated that it might be possible to control cardiac arrhythmias through biofeedback. Studies by Elder et al. (1973) have provided some hope that blood pressure in humans might also be conditioned. Also, considerable effort has been directed to the control of responses from single muscles with particular applied emphasis in neuromuscular rehabilitation, control of muscle tension for tension headaches and the management of migraine headaches through vasomotor conditioning (Brudny et al., 1974; Basmajian, 1963, 1971; Sargent et al., 1973).

Biofeedback, Psychology↗
PubMed1998

Evaluation of toxicity indicators in rat primary astrocytes, C6 glioma and human 1321N1 astrocytoma cells: can gliotoxicity be distinguished from cytotoxicity?

A comparison was made of rat primary astrocytes, C6 glioma cells pre-treated with dibutyryl cyclic AMP, and the human astrocyte 132N1 cell line using a range of 40 compounds and the neutral red (NR) assay. The 40 chemicals included substances known to be toxic to astrocytes or neurons, to be generally cytotoxic or not thought to be toxic to nervous tissue. For those compounds which were toxic, changes in glial fibrillary acidic protein (GFAP) levels were measured in the primary and C6 cultures, and changes in vimentin and S-100 measured in the C6 cells. The number of compounds with EC50 values < 2000 microg/ml for the NR assay for the different cell cultures were as follows: primary astrocytes, 19; C6 cells, 15; and 1321N1 cells, 11. The log of the EC50 values for the NR assay for the test compounds between the three cell types was not significantly different at the 5% level by paired Student's t-test. For the toxic substances the correlation coefficients of the EC50 values between primary cells and the C6 or 1321N1 cells were r > 0.5, and between the C6 and 1321N1 cells r > 0.9. For GFAP there was a similar degree of correlation in EC50 values between the different cell types. The GFAP, vimentin and S-100 levels showed similar EC50 values for the toxicants, but were not as sensitive as the NR assay. The toxic substances caused altered morphology in the primary, C6 and 1321N1 cells, with increased branching of cell processes. The combined astrocyte systems identified 8 out of 9 substances reported to be toxic to astrocytes in vivo, together with substances which have general cytotoxic properties. A number of substances (including the 1 out of 9 reported gliotoxic substances), which may primarily affect neurons, which may affect nervous tissue after long-term exposure, or which are not thought to be toxic to nervous tissue, were not detected. The astrocyte systems positively identify gliotoxic and cytotoxic substances and will allow detailed mechanistic studies to be made on the different underlying mechanisms.

Animals↗
PubMed1999

Collecting duct morphogenesis.

The collecting duct system of the metanephric kidney develops from the ureteric bud, an outgrowth from the caudal end of the Wolffian duct. The ureteric bud is induced to form by signals emanating from a specific area of intermediate mesoderm, which it immediately invades. In response to further mesenchyme-derived signals, the ureteric bud ramifies to form a tree-like collecting duct system, which in turn signals clumps of the mesenchyme cells that surround it to differentiate into epithelial nephrons. The morphogenesis of the collecting duct system is driven by two processes--growth and branching--which are to some extent separable. Each depends on diffusible signals, a number of which have been identified in recent years; growth promoters include hepatocyte growth factor and activin, while ramogens include glial cell line-derived neurotrophic factor, neurturin and persephin. Arborisation also depends on matrix components, including proteoglycans, integrins and their ligands, and metalloproteinases, such as matrix metalloproteinase-9, that are involved in matrix remodelling. So far, little progress has been made in elucidating the intracellular pathways responsible for translating growth factor "instructions" into morphological change, but a role for some intracellular components, such as protein kinase C, formins and the cytoskeleton, is implied by recent experimental data. More information on these internal pathways of control is expected over the next few years.

Aging↗