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At least 793 records · Page 44Linked to original sources
PubMed2006

Proliferation and differentiation of neural stem cells on lysine-alanine sequential polymer substrates.

The purpose of this study was to explore the phenotypic potential of embryonic rat cerebral cortical stem cells by inducing differentiation on lysine-alanine sequential (LAS) polymer substrates at neurosphere level. LAS polymer is a heterologous polymer of lysine and alanine and has been demonstrated to enhance axon growth of neurons in a serum-free medium in vitro. It was found that very few cells migrated outside of the neurospheres but extremely long processes extended from differentiated cells could form a network between remote neurospheres when cells were cultured on LAS substrates at a low density of 120 neurospheres/cm(2) in the serum-free medium. On the contrary, when the neurosphere density was increased to 360 neurospheres/cm(2), many neurosphere-forming cells migrated out from their original aggregate and exhibited short processes morphology. Furthermore, when serum was added to the culture system, the neurosphere-forming cells could be induced into an extensive cellular substratum of protoplasmic cells upon which process-bearing cells spread. Clearly, neurospheres could exhibit different behaviors on LAS substrates according to the complex environmental conditions. Here, we proposed that neurospheres would change their social communication and adopt different strategies to communicate with other neurospheres when they detected each other's presence. Therefore, the mediation of cell behavior on LAS substrates by communication between neurospheres should be taken into account.

Animals↗
PubMed2006

The effect of chitosan and PVDF substrates on the behavior of embryonic rat cerebral cortical stem cells.

In this study, the behavior of neural stem cells from embryonic rat cerebral cortex were compared on the chitosan and poly(vinylidene fluoride) (PVDF) substrates at single-cell and neurosphere level. It was found that chitosan and PVDF substrates inhibited the proliferation and differentiation of single neural stem cells. It seemed that single-cell cultures on both substrates show cells remained dormant. However, neurospheres could exhibit different or similar behavior on these two substrates, which is dependent on the presence or absence of serum. More cells migrated outside from the neurospheres and longer processes extended from differentiated cells on chitosan than on PVDF when neurospheres were cultured in the serum-free medium. On the contrary, when serum was added to the culture system, chitosan and PVDF could induce the neurosphere-forming cells into an extensive cellular substratum of protoplasmic cells upon which process-bearing cells spread. In addition, based on the immunocytochemical analysis, the percentages of differentiated cell phenotypes of neurospheres cultured on chitosan and PVDF substrates became similar in the presence of serum. Therefore, it is reasonable to suggest that biomaterials may stimulate or inhibit the proliferation and differentiation of neural stem cells according to the complex environmental conditions. The information presented here should be useful for the development of biomaterials to regulate the preservation, proliferation, and differentiation of neural stem cells.

Animals↗
PubMed2004

Schwann cell and epineural fibroblast expression of serine racemase.

D-serine appears to be a natural agonist at the "glycine site" of the NMDA receptor and is created by conversion from L-serine by serine racemase. This racemase has been localized to protoplasmic astrocytes that ensheath synapses and modulate neuronal activity in the CNS, but serine racemase expression in the PNS has not been reported. Immunofluorescence indicated that Schwann cells and other endoneurial components of rat spinal nerve contain serine racemase, and western blot analysis detected the enzyme in lysates of sciatic nerve. Cultures from sciatic nerve contained Schwann cells and fibroblasts, and both cell types showed serine racemase expression by immunofluorescence and Western blot; the quantities per unit of total protein appeared slightly lower than that expressed in cultured astrocytes. Cultures enriched for each cell type were subjected to reverse transcriptase polymerase chain reaction, further confirming serine racemase mRNA in Schwann cells and fibroblasts. Finally, immunodetection of D-serine itself was observed in cultured Schwann cells and fibroblasts. These expression patterns of serine racemase may indicate roles for D-serine in peripheral nerve transduction.

Animals↗
PubMed2006

Neuron theory, the cornerstone of neuroscience, on the centenary of the Nobel Prize award to Santiago Ramón y Cajal.

Exactly 100 years ago, the Nobel Prize for Physiology and Medicine was awarded to Santiago Ramón y Cajal, "in recognition of his meritorious work on the structure of the nervous system". Cajal's great contribution to the history of science is undoubtedly the postulate of neuron theory. The present work makes a historical analysis of the circumstances in which Cajal formulated his theory, considering the authors and works that influenced his postulate, the difficulties he encountered for its dissemination, and the way it finally became established. At the time when Cajal began his neurohistological studies, in 1887, Gerlach's reticular theory (a diffuse protoplasmic network of the grey matter of the nerve centres), also defended by Golgi, prevailed among the scientific community. In the first issue of the Revista Trimestral de Histología Normal y Patológica (May, 1888), Cajal presented the definitive evidence underpinning neuron theory, thanks to staining of the axon of the small, star-shaped cells of the molecular layer of the cerebellum of birds, whose collaterals end up surrounding the Purkinje cell bodies, in the form of baskets or nests. He thus demonstrated once and for all that the relationship between nerve cells was not one of continuity, but rather of contiguity. Neuron theory is one of the principal scientific conquests of the 20th century, and which has withstood, with scarcely any modifications, the passage of more than a 100 years, being reaffirmed by new technologies, as the electron microscopy. Today, no neuroscientific discipline could be understood without recourse to the concept of neuronal individuality and nervous transmission at a synaptic level, as basic units of the nervous system.

Animals↗
PubMed2004

Cytoplasmic streaming in plants.

Plant cells are surrounded by a cell wall composed of polysaccharides and hence can change neither their form nor their position. However, active movement of organelles (cytoplasmic streaming or protoplasmic streaming) is observed in plant cells, and involvement of the actin/myosin system in these processes has been suggested. Successful biochemical and biophysical approaches to studying myosins have extensively promoted the understanding of the molecular mechanism underlying these phenomena.

Actin Cytoskeleton↗
PubMed2005

Hypothalamic agouti-related protein immunoreactivity in food-restricted, obese, and insulin-treated animals: evidence for glia cell localization.

Agouti-related protein (AGRP) has been implicated in the regulation of metabolic balance. Overexpression of this peptide leads to obesity. Its activity is mediated via the melanocortin-4 (MC-4) receptor where it acts as an MC-4 receptor antagonist. In this study, we characterized the AGRP brain distribution and cellular localization in control, food-restricted, obese, and insulin-treated rats using immunohistochemistry. AGRP immunostaining was found selectively in regions of the arcuate and ventromedial hypothalamic nuclei. These regions were stained less intensely in food-restricted rats than in controls. AGRP-positive cells in the hypothalamus of obese animals were three times more numerous than in control rats. Also, insulin treatment acted to decrease AGRP immunostaining. Analysis of AGRP cellular localization demonstrated its presence in the cytoplasm of numerous small (7-12 microm) cell bodies of putative protoplasmic astrocytes as well as in nerve fibers. Glia fibrillary acidic protein (GFAP) immunostaining of sections adjacent to those stained for AGRP revealed astrocytes with morphology similar to AGRP-positive cells. A few AGRP-positive nerve cell bodies were also found in the arcuate nucleus of obese rats. We conclude that AGRP hypothalamic content is decreased by fasting and intracerebroventricular (i.c.v.) insulin treatment and increased in obesity. In addition to its presence in nerve fibers, AGRP localization in astroglia-like cells suggests a possible role for these elements in its synthesis or its sequestration from the neuronal compartment.

Agouti-Related Protein↗
PubMed2005

Ballistic labeling and dynamic imaging of astrocytes in organotypic hippocampal slice cultures.

Protoplasmic astrocytes in mammalian CNS tissues in vivo have a highly complex 3D morphology, but in dissociated cell cultures they often assume a flattened, fibroblast-like morphology bearing only a few, simple processes. By fluorescent labeling and confocal reconstruction we show that many astrocytes in organotypic hippocampal slice cultures exhibit a more native complex cytoarchitecture. Although astrocytes at the surface of slice cultures show a reactive form with several thick glial fibrillary acidic protein (GFAP)-positive processes, astrocytes situated in deeper portions of tissue slices retain a highly complex 3D morphology with many fine spine- or veil-like protrusions. Dozens of astrocytes can be labeled in single slice cultures by gene gun-mediated ballistic delivery of gold or tungsten particles carrying cDNAs (Biolistics), lipophilic dyes (DiOlistics), or fluorescent intracellular calcium indicators (Calistics). Expression of a membrane-targeted form of eGFP (Lck-GFP) is superior to soluble eGFP for resolving fine astrocytic processes. Time-lapse confocal imaging of Lck-GFP transfected astrocytes or "calistically" labeled astrocytes show structural remodeling and calcium transients, respectively. This approach provides an in vitro system for investigating the functional architecture, development and dynamic remodeling of astrocytes and their relationships to neurons and glia in live mammalian brain tissues.

Animals↗
PubMed2004

Hypotheses for the functions of intercellular bridges in male germ cell development and its cellular mechanisms.

In oogamous reproduction of multicellular organisms, a striking phenomenon is the prevailing synchronous development of male germ cells connected by wide intercellular bridges (IBs, 0.1-2 microm), which is well conserved in both animal and plant species ranging from algae to human. In the literature, IBs are believed either to allow genetically segregated haploid spermatids to share diploid gene products after meiosis, or to mediate rapid transfer of some vital signals or nutrients. Although intercellular sharing of gene transcripts has experimental evidences, these hypotheses are still not satisfactory. To explore the unknown roles of IB, we assume that developing male germ cells may be especially sensitive to stochastic gene expression to become heterogeneous. To achieve best gamete quality, such heterogeneity must be eliminated so that relatively uniform gametes with normal functions can be produced. Development within a common syncytium may be the only way for this purpose. The process may require not only the intercellular exchange of a few molecular signals but also the mixing of protoplasm between the connected cells so that they have similar levels/states of mRNAs, proteins and organelles, which can be achieved only through wide IBs. This hypothesis can explain some quite intriguing aspects of male gametogenesis and provide unique predictions that can be tested experimentally.

Animals↗
PubMed2004

In vitro differentiated neural stem cells express functional glial glutamate transporters.

The possibility to isolate stem cells from the adult central nervous system and to maintain and propagate these cells in vitro has raised a general interest with regards to their use in cell replacement therapy for degenerative brain diseases. Considering the critical role played by astrocytes in the control of glutamate homeostasis, we have characterised the expression of functional glutamate transporters in neural stem cells exposed to selected culture conditions favouring their differentiation into astrocytes. Commonly, neural stem cells proliferate in suspension as neurospheres in serum-free medium. The addition of serum or a supplement of growth factors (G5) to the culture medium was found to trigger cell adhesion on coated surfaces and to favour their differentiation. Indeed, after 7 days in these conditions, the vast majority of the cells adopted markedly distinct morphologies corresponding to protoplasmic (with serum) or fibrous (with G5 supplement) astrocytes and approximately 35-40% acquired the expression of the glial fibrillary acidic protein (GFAP). Immunocytochemical analysis also revealed that the treatments with serum or with the G5 supplement triggered the expression of the glial glutamate transporters GLT-1 (35 and 21%, respectively) and GLAST (29 and 69%, respectively). This effect was correlated with a robust increase in the Na+ -dependent [3H]-d-aspartate uptake, which was partially inhibited by dihydrokainate, a selective blocker of GLT-1. Together, these results indicate that in vitro differentiation of cultured neural stem cells can give rise to distinct populations of astrocytes expressing functional glutamate transporters.

Amino Acid Transport System X-AG↗
PubMed2003

A transgenic mouse model for the detailed morphological study of astrocytes.

We have generated a transgenic mouse model in which astrocytes express an enhanced green fluorescent protein (EGFP) under the control of the mouse glial fibrillary acidic protein (GFAP) promoter. EGFP, which is characteristically found throughout the cell, was expressed in these animals even in astrocytic fine processes, and EGFP expressing cells demonstrated morphological characters of protoplasmic, fibrous, or reactive astrocytes. In contrast, GFAP immunoreactivity was found only in the perinuclear region and in the main processes. The transgenic mouse model therefore provides a valuable tool for the detailed morphological investigation of astrocytes.

Animals↗
PubMed2002

Behavior of silkworm yolk protein on phospholipid membranes.

During early development, the plasma membrane of silkworm (Bombyx mori) eggs undergoes a superficial cleavage that separates the blastodermal protoplasm and the yolk. To test whether the blastoderm absorbs yolk through the plasma membrane in B. mori, we studied the interaction of phospholipid membranes and yolk using a phospholipid planar bilayer membrane (PBM) and liposomes. In addition, egg-specific protein (ESP; 225 kDa), a yolk protein that is specific to B. mori eggs, was collected by fractionating the eggs. Liposomes were mixed with either B. mori yolk or ESP, and observed under an electron microscope. This showed that the phospholipid membrane was spanned by fine particles 10-20 nm in diameter. Both yolk and ESP caused the PBM to become extraordinarily leaky, with a membrane potential of -70 mV for yolk and -198 mV for ESP. These results suggest that although it is a water-soluble protein, ESP permeates the phospholipid membrane without the help of enzymes.

Animals↗
PubMed1985

Effects of mercaptans upon dihydropyridine binding sites on transverse tubules isolated from triads of rabbit skeletal muscle.

The binding of nitrendipine to transverse (T) tubules isolated from skeletal muscle triads is inhibited by dithiothreitol (KI approximately 0.05 mM) and glutathione (KI approximately 3 mM). The t 1/2's of inhibition (18.3 and 11.5 min, respectively) suggest that these hydrophylic reagents act upon the exposed surface of the vesicles. Dithiothreitol shifts the apparent KD for nitrendipine from 8.5 nM to 30 nM without altering the Bmax extrapolated by Scatchard analysis. That T-tubules isolated by disruption of triad junctions are constrained to have the protoplasmic (P) face uniformly exposed was experimentally confirmed. These studies show that a sulfhydryl residue on the P-face of the T-tubule influences the affinity of the receptor for dihydropyridines.

Animals↗
PubMed2002

Nerve growth factor signals via preexisting TrkA receptor oligomers.

Nerve growth factor (NGF) promotes neuronal survival and differentiation by activating TrkA receptors. Similar to other receptor tyrosine kinases, ligand-induced dimerization is thought to be required for TrkA receptor activation. To study this process, we expressed TrkA receptors in Xenopus laevis oocytes and analyzed their response to NGF by using a combination of functional, biochemical, and structural approaches. TrkA receptor protein was detected in the membrane fraction of oocytes injected with TrkA receptor cRNA, but not in uninjected or mock-injected oocytes. Application of NGF to TrkA receptor-expressing oocytes promoted tyrosine phosphorylation and activated an oscillating transmembrane inward current, indicating that the TrkA receptors were functional. Freeze-fracture electron microscopic analysis demonstrated novel transmembrane particles in the P-face (protoplasmic face) of oocytes injected with TrkA cRNA, but not in uninjected or mock injected oocytes. Incubating TrkA cRNA-injected oocytes with the transcriptional inhibitor actinomycin D did not prevent the appearance of these P-face particles or electrophysiological responses to NGF, demonstrating that they did not arise from de novo transcription of an endogenous Xenopus oocyte gene. The appearance of these particles in the plasma membrane correlated with responsiveness to NGF as detected by electrophysiological analysis and receptor phosphorylation, indicating that these novel P-face particles were TrkA receptors. The dimensions of these particles (8.6 x 10 nm) were too large to be accounted for by TrkA monomers, suggesting the formation of TrkA receptor oligomers. Application of NGF did not lead to a discernible change in the size or shape of these TrkA receptor particles during an active response. These results indicate that in Xenopus oocytes, NGF activates signaling via pre-formed TrkA receptor oligomers.

Animals↗
PubMed1969

Time-lapse cinemicrographic studies of x-irradiated HeLa S3 cells. II. Cell fusion.

Analysis of time-lapse cinemicrographs of X-irradiated HeLa S3 cells has shown that the incidence of cell fusion was increased from 0.9% (following 1267 divisions) in control cells to an average of 22% (following 655 divisions) in cells irradiated with 500 rad doses of 220 kv X-rays. The incidence depended on the stage of the generation cycle at which the parent cells were irradiated. It was nearly constant in the first three postirradiation generations. Fusion occurred at all stages of the generation cycle, but preferentially during the first 20%. Cells undergoing fusion progressed more slowly through the generation cycle and had a higher probability of disintegrating than did irradiated cells that did not fuse. The occurrence of fusion was clonally distributed in the population. It took place only between sister (or closely related) cells. Protoplasmic bridges were often visible between sister cells prior to fusion. Giant cells arose only as a result of fusion. The incidence of multipolar divisions, though higher than in unirradiated cells, was only 5.5% in cultures irradiated with 500 rads. Fusion occurred following 85% of the multipolar divisions and was often followed by a multipolar division.

Cell Biology↗
PubMed1973

Nuclear magnetic resonance studies of sodium and potassium in etiolated pea stem.

Based on nuclear magnetic resonance (NMR) spectroscopy and other evidence, it has been argued that tissues accumulate, and retain, ions in a binding process by a highly structured water-protoplasm system; thus active membrane transport need not be involved. Recent evidence has accounted for the loss of resonance intensity usually found when investigating quadrupolar ions in animal tissue. Using continuous wave NMR spectroscopy, we have examined two quadrupolar ions, Na(+) and K(+), in pea stem cells where about 90% of the ion content is in the largely aqueous vacuoles having a membrane barrier. The NMR resonances from these ions correspond to almost 100% of that expected from independent measurements of total ion content. This indicates that the ions are retained as free ions after accumulation. The small fraction which is NMR invisible may represent ions in an ordered, anisotropic environment, such as that in the wall or cytoplasm.

Biological Transport↗
PubMed1985

Quantitative characterization of a biological membrane by means of its spatial autocovariance.

Profiles for the exoplasmic face (EF) of the freeze-fractured plasma membrane from the root storage tissue of red beets are reconstructed by microdensitometry of micrographs of surface-shadowed-platinum carbon replicas. Autocovariance functions (ACFs) are computed from those profiles. The initial portions of the ACFs have a Gaussian form whose parameters (root mean square surface roughness and autocovariance length) are estimated. The parameter estimates are used to show that the pits on the EF faces are in good complementarity with the intramembrane particles seen on the complementary protoplasmic fracture faces.

Cell Membrane↗
PubMed2000

Immunocontent and secretion of S100B in astrocyte cultures from different brain regions in relation to morphology.

Primary astrocyte cultures prepared from neonatal hippocampus, cerebral cortex and cerebellum were morphologically distinct. Cells from hippocampus and cortex were almost entirely protoplasmic, whereas cerebellar astrocytes had many processes; in the absence of serum these differences were accentuated. We compared the immunocontent and secretion of the mitogenic protein S100B in these cultures. Immunocontent was 2.5 times higher in cerebellar astrocytes than in hippocampal or cortical astrocytes. Cells from all three regions secreted S100B under basal conditions, but the secretion rate was higher in cerebellar astrocytes. Secretion depended on protein synthesis and was increased by incubation with forskolin or lysophosphatidic acid in mechanisms which were additive. The stellate morphology induced by forskolin was reversed by lysophosphatidic acid in hippocampal but not in cerebellar cultures, suggesting that S100B secretion was not associated with a process-bearing phenotype of astrocytes.

Animals↗
PubMed1997

Correlates of p53- and Fas (CD95)-mediated apoptosis in Alzheimer's disease.

Apoptosis may be an important mechanism of cell loss in Alzheimer's disease (AD). Experimentally, apoptosis is preceded by nuclear accumulation of p53, and increased expression of Fas (CD95) antigen. In the present study, quantitative Western blot analysis of postmortem frontal and temporal lobe tissue demonstrated significantly higher mean levels of p53 and Fas in AD relative to age-matched controls. Immunohistochemical staining and in situ apoptosis assays demonstrated increased p53 and Fas expression and DNA fragmentation in overlapping populations of cortical neurons, and cortical and white matter glial cells distributed in regions damaged by neurodegeneration. Double-label immunohistochemical staining studies revealed p53 immunoreactivity in: 1) cortical neurons without tau-immunoreactive neurofibrillary tangles; 2) numerous, but not all tau-immunoreactive neuropil neurites and white matter axons; 3) dystrophic fibrils surrounding amyloid-beta-immunoreactive plaques; and 4) glial cells characterized as A2B5+ protoplasmic astrocytes or oligodendrocytes. The prominent distribution of dystrophic p53-immunoreactive processes around amyloid-beta-containing plaques suggests that amyloid deposits are associated with local neuritic degeneration. In addition, the results suggest that many tau-immunoreactive neuritic processes originate from degenerating (p53) as well as regenerating neurons. Finally, apoptosis of glial cells (A2B5+) required to maintain the functional integrity of axons and dendrites may represent an important pathogenic mechanism of axonal loss and synaptic disconnection in AD.

Aged↗