SEARCH · Search PubMed
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
Two different types of protoplasmic astrocytes were studied by scanning transmission and conventional electron microscopy in order to determine the exact nature of the aspect described by Chan Palay and Palay as velate astrocyte. This study provides new data about the usefulness of S.T.E.M and brings into question the results of the Palays.
Plasma membranes from neuronal perikarya (N), protoplasmic astrocytes (A) and oligodendrocytes (O) of rat brains were analysed with respect to their protein and glycoprotein contents and specificities. SDS-polyacrylamide gel electrophoresis revealed a total number of 23, 17, and 17 major proteins in N, A, and O respectively. Periodate-Schiff's staining showed that approximately 40-60% of these proteins were glycoproteins. The reactivity of these glycoproteins to Con A and WGA was also studied. Selective iodination of whole cells followed by electrophoresis and autoradiography indicated that of the major proteins, only 25% of neuronal and 60% of astroglial and oligodendroglial membrane proteins were exposed outside the cell surface. The overall results suggest that membrane proteins of each of the three cell types studied here have characteristically different internal and external markers differing in size, glycoprotein content, and reactivity of the glycoproteins to lectins.
The effects of the calmodulin antagonists W-7 and trifluoperazine have been measured on the Ca2+-activated potassium channel in the membrane surrounding protoplasmic drops expressed from internodal cells of charophyte plants. The large-conductance (170 pS), voltage- and Ca2+-dependent gating, and prominent conductance substrate of this channel shows a strong kinetic resemblance to those of the Maxi-K channel from animal cells. This is the first study of the action of calmodulin antagonists which measures their effects on the most populated substates as well as the closed and main open states of Maxi-K channels. The substate analysis provides new evidence for different modes of action of- and different bindings sites for these calmodulin antagonists. Neither antagonist produces the simple closure of the channel reported previously as its effect on the Maxi-K channel, though both do induce flicker-block, reducing the mean current to near zero at high concentrations following an inverted Michaelis-Menten curve. W-7 reduces residence time in the fully open state, thus raising, in the same proportions, the probabilities of finding the channel in the closed state or a pre-existing substate. Its binding to the channel is voltage- and calcium-dependent. In contrast, trifluoperazine reduces residence in the open state and promotes an apparently new substate which overlaps the closed state at -50 mV but is distinguishable from it at voltages more negative than -100 mV. This substate may represent times that trifluoperazine is bound to the channel. Both antagonists have effects clearly distinguishable from that of withdrawing calcium from the channel, which does not affect open state residence time but increases closed state residence time. Thus neither antagonist reverses the activating effect of Ca2-. This is good kinetic evidence against the view that the channel is activated by Ca2+-calmodulin and that the effect of a calmodulin antagonist is to reverse this process by making Ca2--calmodulin less available.
By hybridizing mouse myeloma cells with spleen cells from a BALB/c mouse immunized with the glial cell-rich fraction prepared from an autopsied human brain, we established a hybridoma that produces a monoclonal antibody to protoplasmic astrocytes (PA). The antibody, named PRAS-1, consistently labeled cytoplasm of PA with a granular pattern. In a few cases, the cytoplasmic processes of several astrocytes in gray and white matter were also stained. The immunoreactivity was lost after periodic acid treatment or methylation, showing that the epitope is composed of a carbohydrate. The cytoplasmic reaction was resistant to protease digestion and lost after incubation in an organic solvent, suggesting that a glycolipid is the antigen. On the other hand, the reaction in the processes disappeared upon protease digestion. Ultrastructurally, the immunoreaction was localized to secondary lysosomes. Cross-reactivity was noted on a small number of incidental neurons, corpora amylacea, hepatocytes and esophageal epithelial cells. A long period of formalin fixation did not deteriorate the antigenicity. PRAS-1 was demonstrated to detect PA immunohistochemically on paraffin sections, and may be applicable to further investigations into development or neoplasms of human astrocytes.
We present a mathematical model for continuously distributed mechanochemical autooscillations (autowaves) in a protoplasmic strand of Physarum polycephalum. The model is based on a hypothesis of local positive feedback between deformation and contraction of the contractile apparatus. This feedback is mediated through a cell regulatory system whose kinetics involves coupling to mechanical strain. Mathematical analysis and computer simulations have demonstrated that the solutions of the model agree quantitatively with the available experimental data. In particular, hydrodynamic interaction alone, between different sections of the strand via the streaming endoplasm, is capable of inducing the characteristic contractile behavior.
Laser light scattering is shown to be an effective means of obtaining a rapid, objective assessment of dynamic changes in the intact plasmodium of the myxomycete Physarum polycephalum during bidirectional (shuttle) streaming. The motion of material in a 100 mum diameter region of a plasmodial vein was studied by following changes in the autocorrelation function of the fluctuations in the scattered light intensity. The autocorrelation function was recorded at 10 s intervals and analyzed to follow changes in the flow velocity of protoplasm associated with shuttle streaming. Rhythmic velocity changes and a "beating" pattern of velocity maxima were readily observed. In an attempt to locate the site of underlying structural changes in the vein responsible for the changing pattern of flow, the average scattered intensity was separated into components derived from moving and stationary scatterers. Periodic variations in the light intensity due to stationary scatterers are related to the streaming cycle and indicate the occurrence of important structural changes in the vein walls. Two possible interpretations of the data are offered; one involving gross dynamic changes in vein structure, the other involving the formation, contraction, or breakdown of fibrillar material in the vein wall during the streaming cycle.
1. Concentrations of 0.1-10 mM Ouabain do not affect oscillating contractions, when applied externally in physiological solutions. 2. Ouabain has no effect, when applied in solutions of increased sodium- (100 mM) and lowered potassium-concentration (0.1 or zero mM). 3. De novo generation of oscillating contractions in protoplasmic drops is not suppressed in Ouabain-solutions. 4. Biochemical studies of Na-K-ATPase did not show any Ouabain sensitive ATPase-activity. 5. Reaction of veins on high concentrations (300 mosm) is discussed as to be a physical effect. 6. It is concluded that no Na-K-ATPase is engaged in triggering oscillating contraction automaticity.
The lack of the glycine transporter 1 (GlyT1) leads to early postnatal death due to failure of respiratory network activity. Here we demonstrate a segregated expression of GlyT1 on different astroglial cell populations of the ventral respiratory group. In TgN(hGFAP-EGFP) mice a combined immunohistochemical and electrophysiological approach was used to define the cellular expression of GlyT1 in the respiratory network. EGFP-labeled cells with outwardly rectifying current-voltage relationship did not express glycine transporter 1, while GlyT1 was abundantly expressed in mature protoplasmic astrocytes, which are electrophysiologically characterized by a large potassium conductance, a more negative membrane potential and the expression of glutamate transporters. Taken together, the vital capacity for the clearance of extracellular glycine is restricted to a subpopulation of astroglial cells.
In this study we produced polyclonal antisera directed to whole cell, outer sheath, protoplasmic cylinder and axial flagella sonicates of Treponema denticola (ATCC 33521) reference strain. Furthermore, the reactivity of the antisera was determined, using the enzyme-linked immunosorbent assay and immunoblotting techniques. As control antigen, other gram-negative bacteria (Salmonella minnesota, Escherichia coli) and related pathogenic spirochetes (Borrelia burgdorferi and Treponema pallidum) were used. It could be shown that the purified antibodies were specific for Treponema denticola and did not cross-react with the control antigens tested. Interestingly, with one exception, the anti-axial flagella antibody reacted with the flagellin of Treponema pallidum but not with Borrelia burgdorferi flagella. It is intended to use these antisera for the characterization of patient isolates in further studies.
Functional separation of the two signals involved in stimulating immunological responses was achieved through the judicious use of two natural bacterial antigens. Native protoplasmic polysaccharide (NPP) extracted from Escherichia coli was immunochemically identical to the lipopolysaccharide (LPS) extracted from the same organism. However, NPP was not endotoxic, not mitogenic, did not fix complement, and was immunologically independent of T cells. The NPP, which appeared to contain only the antigenic signal, could induce a primary antibody response in mice and could sensitize mice for a secondary response. However, the antigenic signal contained in NPP was insufficient to trigger a secondary response in mice primed with either NPP or LPS. LPS, containing both the antigenic and second signals, was required to trigger a secondary response in primed mice.
A method was devised for estimtaing the electrical resistance between the cytoplasm of adjacent cells of the red marine alga Griffithsia globulifera. A very high resistance was found, which argues strongly against the existence of protoplasmic continuity in this species.
Effects of octylguanidine (OG) were studied on the permeability of cells of the adaxial epidermis of Allium cepa bulb scales to water and methyl urea and on the protoplast surface. Interference of OG with the Ca(2+) and Al(3+) action on the cell surface was also investigated.Permeability of the cell membrane for water and methyl urea increased nearly three times in presence of OG. The effect of OG on cell permeability depended on its direct contact with the protoplast surface.The effect of OG on the interaction between the protoplast surface and the cell wall (wall attachment) was marked and rapid; OG (225 micromolar) decreased the time for protoplast detachment in hypertonic solutions from 420 to 120 seconds. The plasmolyzed protoplasts were immediately rounded off while the controls without OG remained heavily concave.A considerable increase in protoplast detachment time and a decrease in rounding percentage were found when cells were plasmolyzed after pretreatment with AlCl(3) (0.05 molar for 2 minutes). This effect was partially reversed by KCl which was further enhanced by addition of OG.Penetration of OG into the mesoplasm was manifested only after 10 to 15 minutes. Vacuolization and swelling of the protoplasm, fragmentation of the protoplast, and aggregation of the spherosomes, however, were observed only 30 minutes after transfer. No evidence for penetration of OG into the vacuole was found.The results support earlier suggestions that OG acts primarily on the protoplast surface by interacting with membrane proteins as well as with phospholipids. In several aspects, OG acts on the cell surface similarly to a surfactant.
The biophysical properties of voltage-dependent K(+)-channels of protoplasmic droplets of Chara corallina Klein ex Willd., em, R.D.W. were investigated using the tight-seal whole cell method. Two potassium currents were observed in voltage-clamp mode and they can be used to explain the transient membrane potential time course observed in current-clamp mode. The K(+)-channels are identified by the effect of tetraethylammonium chloride which blocks both currents. A two-state, constant dipole moment model is used to fit the voltage-conductance curve. From this model the minimum equivalent gating charge involved in the gating mechanism of K(+)-channels of Chara can be estimated.
The protoplasmic viscosity was studied by using a small spin label having high permeability and broad solubility properties and nickel chloride as an extracellular spin-subtracting agent to localize signal inside cells. The viscosity is variable and in some cells is many times that of water or phospholipids, suggesting that lateral diffusion in biological membranes is important to cell function.
The capacity to precipitate quantitatively with homologous antiserum of native protoplasmic polysaccharide from Escherichia coli was not reduced by incubation in human plasma; it was reduced with the immunochemically related lipopolysaccharide.
Using immunoblots, we identified proteins of Borrelia burgdorferi recognized by sera from 62 patients with either acute or chronic Lyme disease. In all groups studied, the 41-kDa flagellar protein and a relatively minor 93-kDa protein (p93) were the most commonly recognized antigens in patients with acute and chronic disease due to B. burgdorferi. A murine monoclonal antibody (MAb 181.1) was developed against p93, and the antigen was detected by immunoblot analysis in four European and American strains of B. burgdorferi. On two-dimensional gel electrophoresis, p93 had an apparent pI of 6.8. Immunoelectronmicroscopy with MAb 181.1 demonstrated that p93 is located within the protoplasmic cylinder compartment of the organism. The gene encoding p93 was retrieved from a phage expression library. The derived amino acid sequence of p93 confirmed chemical characterization of the antigen, including its amino-terminal peptide sequence. The derived amino acid sequence predicted it to be predominantly alpha helical. A prominent antigenic domain located at the carboxy portion of the protein was recognized by human and rabbit polyclonal antisera and human (MAb D4) and mouse (MAb 181.1) MAbs.
Hyphal tips of fungi representing Oömycetes, Zygomycetes, Ascomycetes, Basidiomycetes, and Deuteromycetes were examined by light and electron microscopy and compared with respect to their protoplasmic organization. In all fungi studied, there is a zone at the hyphal apex which is rich in cytoplasmic vesicles but nearly devoid of other cell components. Some vesicle profiles are continuous with the plasma membrane at the apices of these tip-growing cells. The subapical zones of hyphae contain an endomembrane system which includes smooth-surfaced cisternae associated with small clusters of vesicles. The findings are consistent with the hypothesis that vesicles produced by the endomembrane system in the subapical region become concentrated in the apex where they are incorporated at the expanding surface. Septate fungi (Ascomycetes, Basidiomycetes, and Deuteromycetes) have an apical body (Spitzenkörper) which is associated with growing hyphal tips. In electron micrographs of these fungi, an additional specialized region within the accumulation of apical vesicles is shown for the first time. This region corresponds on the bases of distribution among fungi, location in hyphae, size, shape and boundary characteristics to the Spitzenkörper seen by light microscopy. This structure is not universally associated with tip growth, whereas apical vesicles are widespread among tip-growing systems.
A tuberculin-active glycopeptide containing eight different amino acids and glucose was isolated from the protoplasm of Mycobacterium tuberculosis. A molecular weight of 4,000 to 5,000 was established by Sephadex gel filtration; other analyses showed a peptide to carbohydrate ratio of 9:1. These observations suggest a tentative composition of 3 to 4 residues of glucose, 12 residues each of aspartic and glutamic acids, 3 residues each of lysine, glycine, and serine, and 1 residue each of arginine, threonine, and alanine.