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At least 811 records · Page 45Linked to original sources
PubMed1999

Immunocytochemical studies of aquaporin 4 in the skeletal muscle of mdx mouse.

Immunostainability of anti aquaporin 4 antiserum was investigated in the muscles of dystrophin deficient mdx mice. Western blot analysis showed that the rabbit antiserum against aquaporin 4 reacted with a 28 kDa protein in extracts of normal mouse quadriceps femoris muscles but did not react with the protein in extracts of quadriceps femoris muscles of mdx mice. Immunoperoxidase staining of the muscles from normal and mdx mice revealed the positive immunoreaction at the myofiber surface of normal mice and the negative, or the faint and discontinuous immunostaining at the surface of mdx myofibers. Immunogold electron microscopy disclosed the localization of aquaporin 4 molecules at the myofiber plasma membranes of normal mice and the localization was consistent with that of orthogonal array particles in the protoplasmic face of normal muscle plasma membrane seen in freeze fracture replicas. This study demonstrated that the density of aquaporin 4 molecules was decreased in the muscle plasma membranes of mdx mice, resulting in the faulty function of mdx myofibers.

Animals↗
PubMed1984

The cell in the field of gravity and the centrifugal field.

It appears that the literature and logic that the earth's gravity has been one factor in the limitation of cell size, as well as being an important influence on the diversity of cell types and sizes throughout biological evolution. Analysis of the literature reveals an inverse relationship between the centrifugal force needed for intracellular stratification and cell size. The cells studied ranged in size from approximately 1 mm (amphibian eggs, Pelomyxa) to 0.01 mm (erythrocyte, lymphocyte), and g-forces ranged from about 100 g to 100 000 g respectively. Stratification within cell nuclei and organelles requires even greater forces, presumably because of their smaller size. Extrapolation from centrifugal forces to the force of gravity, and from the full stratification to the initial sedimentation of cell parts suggests a hypothesis for the evolutionary survival and existence of cells in the field of gravity. Average cell size results, in part, from the physical equilibrium between the destructive influence of the force of gravity and the protective role of diffusion and the cytoskeleton. At increased forces of gravity the cell size would thus be decreased, whereas at lower gravitational forces and weightlessness cell size would be expected to increase. Mechanisms of protection of giant cells against internal sedimentation are based on protoplasmic motion, thin and elongated shape of the cell body, increased cytoplasmic viscosity, and a reduced range of specific gravity of cell components, relative to the ground-plasm. The nucleolus, due to its higher density, is considered as a possible trigger of mitosis.

Animals↗
PubMed1983

Reorganization of membrane ergosterol during cell fission events of Candida albicans: a freeze-fracture study of distribution of filipin-ergosterol complexes.

The redistribution of ergosterol molecules which occurs during bud and germ tube formation (dimorphism) in Candida albicans was studied using filipin, a sterol-specific antibiotic, and examined by the freeze-fracture technique. When cells were fixed in a glutaraldehyde solution containing 50 micrograms/ml of filipin, filipin-ergosterol complexes, which were recognized as either pits on the exoplasmic fracture face or protuberances on the protoplasmic fracture face, were homogeneously distributed on the yeast plasma membranes. The plasma membrane of young budding yeast cells demonstrated few filipin-ergosterol complexes compared to the parent yeast plasma membrane. In addition, at a certain time during enlargement of budding yeast cells, the complexes became virtually absent from the constricted region between daughter and parent yeast cell. On the other hand, when germ tubes emerged as cylindrical outgrowths from the parent yeast cells, filipin-ergosterol complexes were heterogeneously redistributed on the plasma membrane. These results suggest that ergosterol molecules may be in lower concentration in the plasma membrane at the constricted region of yeast cell than elsewhere on the plasmalemma of the yeast cell.

Candida albicans↗
PubMed1999

Proteolytic activity in relation to seasonal cambial growth and xylogenesis in Pinus banksiana.

Proteolytic activity in the cambial zone and developing xylem of Pinus banksiana Lamb. was investigated over an annual cycle of growth and dormancy. Highest proteolytic activity was associated with the most active period of primary-wall radial expansion of cambial derivatives, in early spring, before protoplasmic autolysis was initiated in developing earlywood. Three pH maxima of proteolytic activity, near pH 3.0, 6.5 and 9.5, were observed at that time. In general, activities measured at pH values below 7.0 were greater than those determined above pH 7.0 at all stages in the annual cycle, in both cambial zone and developing xylem, although elevated activity at alkaline pH was also observed during springtime growth. Polyvinylpolypyrollidone (PVP) treatment markedly enhanced pH 7.5 but not pH 4.0 proteolytic activity in the cambial zone, but not in developing xylem, indicating the presence of PVP-binding proteinase regulators in the cambium. By fractionation and effector studies total proteolysis was determined to comprise interactions between serine, cystine, aspartate and metallo-proteases having MWs, by gel chromatography, between 10 and 100 kDa. The observations point to a complex regulatory mechanism controlling the presence and catalytic rates of the distinct types of proteases in the cambial region throughout an annual cycle of growth and dormancy.

Cycadopsida↗
PubMed2003

Astrocytic cytoplasmic inclusions within an epileptic focus in an otherwise neurologically intact patient.

A 17-year-old woman with intractable seizures since infancy underwent resection of an epileptic focus in the left frontal cortex. The cytoplasm of many cortical astrocytes contained amorphous eosinophilic inclusions, which ultrastructurally were non-membrane bound and consisted of densely packed osmiophilic material. Similar inclusions have previously been observed, at autopsy, in patients with unspecified mental retardation and various brain malformations. The present report is unique in that the inclusions were detected in the resected specimen of an epileptic focus. The patient is neurologically intact except for the seizures that presently are totally controlled by the surgery. The pathogenesis of these inclusions is unclear. The fact they occurred in an epileptic focus raises the possibility that prolonged seizures or its underlying precipitating factors may cause conglutination of an indeterminate element of the protoplasmic astrocytes resulting in inclusion formation.

Adolescent↗
PubMed1981

Visualization of calcium-binding sites at plasma membrane of shock-frozen Acanthamoeba cells.

Electron-dense deposits appear at the protoplasmic side of plasma membrane in Acanthamoeba log-phase cells when fixed either with glutaraldehyde or formaldehyde supplemented with Ca2+. Similar deposits appear when the cells are preloaded with Ca2+ and thereafter shock-frozen and prepared for electron microscopic examination by freeze-substitution technique. This suggests that their formation reflects the presence of a physiologically active system involved in capturing excess of inflowing calcium.

Amoeba↗
PubMed1989

Electron microscopical localization of guanylate cyclase activity in the neocortex of the guinea pig.

The localization of the guanylate cyclase (GC) activity has been established in the neocortex of adult guinea pigs by means of electron microscopical histochemistry [the DMSO-method of Fujimoto et al. (1981)]. Reaction product was deposited within a population of large- and medium-sized cortical neurons as well as in the cytoplasm of a part of the dendrites of variable size and in the cytoplasm and the nuclear membrane of a number of protoplasmic astrocytes. In the perikarya of the positive neurons, the reaction precipitate was mainly located within the cisterns of the rough endoplasmic reticulum and on the nuclear membrane. In the dendrites, the reaction product was usually distributed in close contact with microtubules, microfilaments, and beneath the postsynaptic membranes of a number of axodendritic synaptic contacts. The axons and all presynaptic boutons were negative. Thus, the localization of the GC could be determined as exclusively postsynaptic. The results obtained support the view for the probable participation of cyclic GMP in the cholinergic, glutaminergic or GABAergic, or peptidergic transmitter mechanisms in the central nervous system.

Animals↗
PubMed1988

Membrane oligosaccharides: structure and function during differentiation.

Recent results gathered by normal light microscopy, immunocytochemistry, fluorescent-analog cytochemistry, and electron microscopy have allowed an improved interpretation of ameboid movement and related phenomena. 1. The contractile system responsible in Amoeba proteus for the generation of motive force for protoplasmic streaming and a large variety of dynamic activities is represented mainly by a thin cortical filament layer at the cytoplasmic face of the cell membrane (Fig. 18I). During normal locomotion this layer exhibits a distinct structural and physiological polarity with three different zones: a zone of reformation at the front (A), a zone of contraction in the intermediate cell region (B), and a zone of destruction at the uroid (C). 2. Two types of filaments participate in the formation of the cortical layer: (1) randomly distributed thin (actin) filaments exhibiting a parallel orientation in the anterior (Fc1) and a disordered arrangement in the intermediate and posterior cell region (Fc2; see also Fig. 17b), and (2) thick (myosin) filaments in close association with F-actin and mostly restricted to the intermediate and posterior cell region (Fc2). 3. The internal hydraulic pressure generated by localized active contraction of the cortical layer is transmitted to the endoplasm via the cell membrane and converted into directed streaming by a gel-sol gradient of decreasing viscosity between the uroid and the front. Calcium ions, ATP, and regulative proteins (profilin and a kinase) play an essential role in controlling both the interaction of actin and myosin and the sol-gel state of the cytoplasmic matrix. 4. Any alterations externally induced in the polarity of the cortical filament system by chemical or physical stimulation and inhibition cause immobilization of the amebas (Fig. 18II) with characteristic changes in (1) cell shape (spherulation and cell flattening), (2) membrane dynamics (cytotic and cytokinetic activities), and (3) cytoplasmic organization (hyalogranuloplasmic separation). pseudopodial tip (Fig. 18III, b----c, d----e), (3) destruction of the old layer at the hyalogranuloplasmic border (Fig. 18III, c,e), and (4) alternate solation (Fig. 18III, b and d) and gelation (Fig. 18III, c and e) of the hyaloplasm between the layer and the plasma membrane. The retraction of pseudopodia is accomplished by a local contraction of the cortical layer in conjunction with a simultaneous gel-sol transformation of the ectoplasmic cylinder. 6. The expression of a rather complex cytoskeleton which is composed not only of microfilaments and associated proteins, but also of intermediate- and microtubularlike structures has to be considered in future

Aging↗
PubMed1998

An improved approach to freeze-fracture morphology of monolayer cell cultures.

Much work is currently done on cell cultures to elucidate membrane processes associated with different cell functions. We describe here a modified freeze-fracture method to obtain systematically large fractured areas of the plasma membrane from monolayer cell culture in situ. Cells are grown until confluence on a Thermanox coverslip overlaid with poly-L-ornithine. After chemical fixation, the culture is flattened overnight by sandwiching it between the Thermanox coverslip, a Falcon membrane and a glass coverslip, under a 5 g weight. After freeze-fracture, vast pictures of the protoplasmic leaflets are obtained in a reproducible manner. Our approach was applied to cultures which were stimulated to release acetylcholine; it has been found very appropriate for studying modifications affecting intramembrane particles and vesicles openings in the plasmalemma. Accurate quantifications were performed and correlations were established between the membrane changes and the data revealed by thin sections. The present sandwich method can be applied to a variety of cell preparations, allowing for quantitative study of structure-function relationships.

Animals↗
PubMed1999

Differential ontogenetic patterns of levocabastine-sensitive neurotensin NT2 receptors and of NT1 receptors in the rat brain revealed by in situ hybridization.

The postnatal ontogeny of the levocabastine-sensitive neurotensin receptor (NT2) mRNA was studied by in situ hybridization in the rat brain and compared with the distribution of the levocabastine-insensitive NT1 receptor. NT2 receptor mRNA was absent at birth from all brain structures except the ependymal cell layer lining the ventricles. The development of NT2 receptor mRNA followed three ontogenetic patterns. The first pattern, involving the majority of the cerebral gray matter, was characterized by a continuous increase from postnatal day 5 (P5) to P30. The second one, involving regions rich in myelinated fibers such as the corpus callosum and lacunosum moleculare layer of the hippocampus, exhibited a pronounced increase between P5 and P10, peaked at P15 and was followed by a plateau or a slight decrease. The third pattern was observed in the ependymal cell layer lining the olfactory and lateral ventricles, where the high labeling already present at birth continued to increase during development. These different developmental patterns could reflect the variety of cells expressing NT2 receptor mRNA, including neurons, protoplasmic astrocytes in gray matter, fibrous astrocytes present in myelinated fibers tracts, and ependymal cells. In contrast, NT1 receptor mRNA, which seems to be associated only with neurons, was highly and transiently expressed during the perinatal period in the cerebral cortex, hippocampus and striatal neuroepithelium. Other regions, notably the ventral tegmental area and substantia nigra compacta, exhibited a gradual increase in NT1 receptor signal, reaching adult levels by P21. Both the differential localization and ontogenetic profiles of NT1 and NT2 receptor mRNAs suggest different involvement of these two receptors in brain functions and development.

Amygdala↗
PubMed1984

Immunocytochemical characterisation of cell cultures grown from dissociated 1-2-day post-natal rat cerebral tissue. A developmental study.

A range of cell-specific markers have been employed with immunocytochemical methods to characterise and quantitate the cell types present in mixed brain cell cultures derived from dissociated 1-2-day post-natal rat cerebral hemispheres and grown in the presence of FCS. Protoplasmic astrocytes (GFAP+, A2B5-) were the major cell type to develop in culture, a confluent monolayer forming in 5-8 days. A population of smaller round cells of oligodendrocyte-like morphology appeared on this astrocyte layer. Greater than 70% of these smaller cells were GC- and thus were not oligodendrocytes. The GC- cells were A2B5+ and, in early cultures, may therefore be progenitor glial cells. Examination of GFAP and A2B5 co-expression by these smaller cells was difficult due to the dense underlying GFAP+ astrocyte layer. In less dense areas of older cultures these smaller cells with processes were GFAP+ and A2B5+: these are Type 2, fibrous astrocytes. GC+ oligodendrocytes, comprising 5-10% of the total identified cell population, were initially distributed over the astrocyte monolayer; in older cultures (after about 8 days) GC+ cells were observed in clumps over places where NF+ cells were identifiable. Such GC+ cells mostly became MBP+. Neurones accounted for about 6% of the identifiable cells in early cultures but a lower percentage in older cultures. Minor populations of ependymal cells and macrophages were present; cells displaying fibronectin, fibroblasts, were rarely identified. Use of horse serum in place of FCS gave lower yields of GC+ cells in cultures, slowed down astrocyte development, and resulted in the formation of trunks of GFAP+ cells throughout cultures. Other sera gave lower numbers of GC+ cells.

Animals↗
PubMed1985

Glial fibrillary acidic protein (GFAP): the major protein of glial intermediate filaments in differentiated astrocytes.

The glial fibrillary acidic protein (GFA protein or GFAP) is the major protein constituent of glial intermediate filaments in differentiated fibrous and protoplasmic astrocytes of the central nervous system. Proteins having similar molecular weights, isoelectric points, and immunoreactivity with GFAP have been found in cells of neural crest and ectodermal origin. A putative function ascribed to glial filaments is its role as a component of the cytoskeleton in defining and maintaining the shape of the astrocyte. Since 1980, over 350 reports have utilized antisera to GFAP for immunochemical and immunocytochemical studies.

Animals↗
PubMed1998

Early, middle, and late stages of neural cells from ovine embryo in primary cultures.

The utilization of neural cells in culture has importantly increased the knowledge of the nervous system biology. In most studies, the investigations are performed on biological materials coming from common laboratory animals and the extrapolation of the results to other animals is not easy. For some studies, such as developmental biology of the nervous system, prion disease investigations, or agronomical production, the utilization of ovine neural cell cultures presents many advantages. Unfortunately, there are few data on the conditions of culture of such cells. In the present work, we investigated simple ways to obtain neurons and astrocytes from sheep brain. Viable neuronal cell cultures were obtained from 40 to 50 day old fetuses. Their morphologies were quite similar to that of neurons from rodent or chick brain and they were labeled by antineurofilament antibodies. Stages older than 50 days of pregnancy were unable to give viable culture of neurons. The stages of 40 day old fetus to newborn lamb were able to give viable astrocyte cultures. The common protoplasmic astrocytes were obtained and they were labeled by antiglial fibrillary acidic protein antibodies. The astrocytes contained glycogen, thus looking like the common astrocytes from rodents. Neuronal or astroglial cultures can be derived from 26 day old embryos, but the cultures contained contaminating cells. Among the latter cells, there were undifferentiated cells which were flat and epitheloid and which were grouped as islets. These cells could be maintained in culture for a time duration over 7 months, even after two passages. They differentiated principally in astrocytes with a radial configuration. This work shows how some neural cells can be simply and easily cultured from sheep brain. For the first time, neurons were cultured from the sheep embryonic brain. Moreover, stem cells were cultured for more than 7 months and, finally, glycogen accumulation in sheep astrocytes was shown to be the same as that in rodent astrocytes. The oligodendrocyte culture was already documented. Thus, sheep can easily be used as well as other models for neural cell studies.

Animals↗
PubMed1987

Freeze fracture study of the schizonts and merozoites of Leucocytozoon caulleryi.

The fine structure of the schizonts and merozoites of Leucocytozoon caulleryi was studied by the freeze fracture technique. Young schizonts were spherical, surrounded by a thick fibrous capsule and possessed numerous nuclei. The nucleus was surrounded by the outer and inner nuclear membrane. A number of nuclear pores were evident in the nuclear membranes. Advanced schizonts were characterized by the formation of cytomeres. Each cytomere was surrounded by a membrane. Several nuclei lay along the periphery of the cytomere. The protoplasmic face (P-face) of the membrane of the cytomere contained numerous intramembranous particles (IMP). The exoerythrocytic merozoites were surrounded by a two-layered pellicle consisting of the outer and inner membrane. During freeze fracturing, each of the two membranes could split along its hydrophobic interior to yield a total of four fracture faces. The P-faces of the outer and inner membranes were closely covered with evenly distributed IMP. The external faces (E-face) of these membranes had less IMP than the P-faces. The membrane architecture of the erythrocytic merozoites was similar to that of the exoerythrocytic merozoites. The erythrocytic merozoite was enveloped by an additional membrane probably derived from the plasma membrane of host cell.

Animals↗
PubMed2002

Neurobiology through the looking-glass: D-serine as a new glial-derived transmitter.

D-Amino acids have been known to be present in bacteria for more than 50 years, but only recently they were identified in mammals. The occurrence of D-amino acids in mammals challenge classic concepts in biology in which only L-amino acids would be present or thought to play important roles. Recent discoveries uncovered a role of endogenous D-serine as a putative glial-derived transmitter that regulates glutamatergic neurotransmission in mammalian brain. Free D-serine levels in the brain are about one third of L-serine values and its extracellular concentration is higher than many common L-amino acids. D-Serine occurs in protoplasmic astrocytes, a class of glial cells that ensheath the synapses and modulate neuronal activity. Biochemical and electrophysiological studies suggest that endogenous D-serine is a physiological modulator at the co-agonist site of NMDA-type of glutamate receptors. We previously showed that D-serine is synthesized by a glial serine racemase, a novel enzyme converting L- to D-serine in mammalian brain. The enzyme requires pyridoxal 5'-phosphate and it was the first racemase to be cloned from eucaryotes. Inhibitors of serine racemase have therapeutic implications for pathological processes in which over-stimulation of NMDA receptors takes place, such as stroke and neurodegenerative diseases. Here, we review the role of endogenous D-serine in modulating NMDA neurotransmission, its biosynthetic apparatus and the potential usefulness of serine racemase inhibitors as a novel neuroprotective strategy to decrease glutamate/NMDA excitotoxicity.

Animals↗
PubMed2002

Whatever happened to the 'microtrabecular concept'?

Keith Porter culminated his stellar career as the founding father of biological electron microscopy by acquiring, in the late 1970s, a high-voltage electron microscope (HVEM). With this magnificent instrument he examined whole-mounts of cultured cells, and perceived within them a structured cytoplasmic matrix he named the "microtrabecular lattice". Over the next decade Porter published a series of studies, together with a team of outstanding young colleagues, which elaborated his broader "microtrabecular concept." This concept posited that microtrabeculae were real physical entities that represented the fundamental organization the cytoplasm, and that they were the physical basis of cytoplasmic motility and of cell-shape determination. The present review presents Porter's original images of microtrabeculae, after conversion to a more interpretable "digital-anaglyph" form, and discusses the rise and fall of the microtrabecular concept. Further, it explains how the HVEM images of microtrabeculae finally came to be considered as an artifact of the preparative methods Porter used to prepare whole cells for HVEM. Still, Keith's "microtrabecular concept" foretold of our current appreciation of the complexity and pervasiveness of the cytoskeleton, which has now been found by more modern methods of EM to actually be the fundamental organizing principle of the cytoplasmic matrix. During the impending eclipse of Porter's microtrabecular concept in the late 1980s, many of Keith's colleagues fondly described the cell as being filled, not with protoplasm, but with "Porterplasm." Despite the fact that Keith's view was clouded by the methods of his time, it would be fitting and apt to retain this name, still today, for the ordered matrix of cytoskeletal macromolecules that exists in the living cell. In the end, the story of what happened to Porter's microtrabecular concept should be an object lesson in scientific hubris that should humble and inform all of us in cell biology, even today--particularly when we begin to think that our most recent methods and observations are achieving "the last word".

Animals↗
PubMed1997

Astrocytic neurotransmitter receptors in situ and in vivo.

In the brain, astrocytes are associated intimately with neurons and surround synapses. Due to their close proximity to synaptic clefts, astrocytes are in a prime location for receiving synaptic information from released neurotransmitters. Cultured astrocytes express a wide range of neurotransmitter receptors, but do astrocytes in vivo also express neurotransmitter receptors and, if so, are the receptors activated by synaptically released neurotransmitters? In recent years, considerable efforts has gone into addressing these issues. The experimental results of this effort have been compiled and are presented in this review. Although there are many different receptors which have not been identified on astrocytes in situ, it is clear that astrocytes in situ express a number of different receptors. There is evidence of glutamatergic, GABAergic, adrenergic, purinergic, serotonergic, muscarinic, and peptidergic receptors on protoplasmic, fibrous, or specialized (Bergmann glia, pituicytes, Müller glia) astrocytes in situ and in vivo. These receptors are functionally coupled to changes in membrane potential or to intracellular signaling pathways such as activation of phospholipase C or adenylate cyclase. The expression of neurotransmitter receptors by astrocytes in situ exhibits regional and intraregional heterogeneity and changes during development and in response to injury. There is also evidence that receptors on astrocytes in situ can be activated by neurotransmitter(s) released from synaptic terminals. Given the evidence of extra-synaptic signaling and the expression of neurotransmitter receptors by astrocytes in situ, direct communication between neurons and astrocytes via neurotransmitters could be a widespread form of communication in the brain which may affect many different aspects of brain function, such as glutamate uptake and the modulation of extracellular space.

Animals↗
PubMed2000

Comparison of astrocytic and myocytic metabolic dysregulation in apolipoprotein E deficient and human apolipoprotein E transgenic mice.

The accumulation of tubular aggregates in type II skeletal muscle fibres and fibrillo-granular inclusions in hippocampal protoplasmic astrocytes are characteristic lesions of apolipoprotein E deficient mice. Moreover these inclusions reacted immunocytochemically with an antibody specific to fragment 17-24 of the published sequence of Alzheimer's amyloid peptide. In an effort to evaluate the role of apolipoprotein E in the formation of these abnormal structures, we examined the tibialis anterior muscle and the hippocampus of several groups of animals including: (i) apolipoprotein E "knockout" mice which had been whole body irradiated with 1200 rads and bone marrow replenished with apolipoprotein E sufficient marrow; and (ii) three transgenic murine strains that had been genetically engineered to express either human apolipoprotein E2, E3 or E4 protein on an apoE deficient background. The results of this study showed that the presence of murine apolipoprotein E (even in subnormal levels in the serum) in irradiated bone marrow replenished mice and in all three (E2, E3 or E4) human apoE transgenic strains was sufficient to prevent the aggregation of sarcoplasmic tubules in the tibialis anterior type II muscle fibres. Similarly apolipoprotein E "knockout" bone marrow replenished mice and all three transgenic strains expressing the different human apolipoprotein E alleles reduced the number of the astrocytic inclusions in the hippocampus to levels not significantly different to those observed in control C57Bl6J animals. The data obtained in this study indicate that neurological and neuromuscular abnormalities found in apoE deficient mice are reversed when apoE protein is replaced in the circulation, either by bone marrow transplantation of normal apoE sufficient marrow, or by gene therapy with the apoE gene, albeit of human origin and irrespective of the allele used.

Alzheimer Disease↗