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PubMed1980

Electron microscopy of the spherical body of oral spirochetes in vitro. Further studies.

The surface and inner structure of the spherical bodies (SB) produced by the human oral treponeme strain G7201, similar to Treponema macrodentium, were studied by electron microscopy. Ultrathin sectioning and scanning techniques demonstrated that in the presence of a high concentration of sucrose, the outer envelope of one or both terminal ends of this oral spirochete changed into a swollen structure, the SB. Spirochetal cells adhered firmly to the surface of the resultant body. The membrane of the SB, i.e. the outer envelope, enclosed the coiled protoplasmic cylinder and five axial fibrils which were located between the envelope and th cylinder. Large expanded protoplasmic cylinders were observed, surrounded by a partially disrupted double membrane in some SBs. A number of frizzly fibrous structures, which differed from axial fibrils in number and shape, were also observed within these SBs. Except for abnormal or partially broken cylinders, the protoplasmic cylinders tended to be located close to the inner surface of the SB membrane, resulting in a central vacant space with occasional axial fibrils. These findings suggest that the oral spirochete produces an SB by terminal expansion of the outer envelope in the presence of high concentrations of sucrose. The outer envelope of the SB, which consists of two electron-dense layers, has the property of binding spirochetal cells to its outer layer and the protoplasmic cylinder and axial fibrils to the inner layer. Some protoplasmic cylinders were also observed to be swollen in the presence of high sucrose concentrations.

Microscopy, Electron↗
PubMed2006

Spherical body formation in the spirochaete Brachyspira hyodysenteriae.

When cultures of Brachyspira hyodysenteriae were grown under a wide range of in vitro conditions, at least 1% of the cells formed spherical bodies different to the normal helical form. This percentage increased considerably in aging cultures or following their incubation in caramelized media. Spherical body formation was initiated from a terminal localized swelling of the outer sheath followed by a retraction of the protoplasmic cylinder into the resulting swollen vesicle. As this occurred, the periplasmic flagella seemed to unwind from the protoplasmic cylinder. Once retracted, the protoplasmic cylinder was found to be wrapped in an organized manner around the inner surface of the membrane of the swollen vesicle. Although most were 2-3 microm in diameter, some much larger spherical bodies (6-12 microm diameter) were occasionally seen, with a corresponding increase in the visible number of peripheral protoplasmic cylinder cross-sections. Spherical bodies from older cultures did not contain protoplasmic cylinders arranged around the periphery, but instead were characterized by the presence of a centrally located, electron-dense body c. 0.5-0.8 mum in diameter. Brachyspira hyodysenteriae spherical bodies differ in both their structural organization and probable method of formation from similar structures described in other spirochaete genera.

Animals↗
PubMed1992

Can we see living structure in a cell?

Colloid chemistry (kappa o lambda lambda alpha: glue, or gelatin) was introduced in 1861 after the discovery of protoplasm which exhibits gelatin-like properties. Some 80 years later, colloid chemistry (and with it, the concept of protoplasm) was largely abandoned. The membrane (pump) theory, according to which cell water and cell solute like K+ are free as in a dilute KCl solution, became dominant. Later studies revealed that rejecting the protoplasmic approach to cell physiology was not justified. Evidence against the membrane (pump) theory, on the other hand, has stood the test of time. In a new theory of the living cell called the association-induction (AI) hypothesis, the three major components of the living cell (water, proteins and K+) are closely associated; together they exist in a high-(negative)-energy-low entropy state called the living state. The bulk of cell water is adsorbed as polarized multilayers on some fully extended protein chains, and K+ is adsorbed singly on beta- and gamma-carboxyl groups carried on aspartic and glutamic residues of cell proteins. Extensive evidence in support of the AI hypothesis is reviewed. From an extension of the basic concepts of the AI hypothesis and the new knowledge on primary structure of the proteins, one begins to understand at long last what distinguishes gelatin from other proteins; in this new light, new definitions of protoplasm and of colloid chemistry have been introduced. With the return of the concept of protoplasm, living structure takes on renewed significance, linking cell anatomy to cell physiology. Finally, evidence is presented showing that electron microscopists have come close to seeing cell structure in its living state.

Animals↗
PubMed1982

Isolation of human erythrocyte inside-out vesicles alters their molecular architecture.

Inside-out (I.O.) vesicles isolated from human erythrocyte ghosts induced to endocytose have been used for biochemical studies to determine localization of molecules within the membrane. It was the purpose of this study to examine such vesicles by freeze-etch electron microscopy to determine the architecture of the peripheral proteins on the protoplasmic surface. Examination of the I.O. vesicles while still in the interior of the ghosts showed that a globular material was randomly distributed on the outer surface (protoplasmic surface of the original plasma membrane) of the I.O. vesicles. The random distribution of the globular material becomes altered, however, if the I.O. vesicles are isolated from the ghosts by shearing and centrifugation. Freeze-etching of these isolated I.O. vesicles revealed that the globular material was now clustered on the protoplasmic surface (PS), as were the intramembranous particles (IMPs) in the extracellular face. Thus, the lateral mobility of the IMPs is dependent upon the distribution of molecules at the protoplasmic surface of the membrane. Moreover, this change in distribution of the globular material at the surface is not due to a partial loss of major membrane proteins, because SDS-polyacrylamide gel electrophoresis revealed that equal amounts of the major proteins were present in nonisolated vesicles as compared to isolated vesicles. Although isolated I.O. vesicles have been used extensively to demonstrate that glycoproteins span the membrane, these results suggest that one should cautiously interpret data obtained from such isolated vesicles in view of the fact that there is an alteration of the distribution and perhaps configuration of molecules at the PS following isolation of I.O. vesicles.

Electrophoresis, Polyacrylamide Gel↗
PubMed1996

Phase switching of oscillatory contraction in relation to the regulation of amoeboid behavior by the plasmodium of Physarum polycephalum.

The plasmodium of the true slime mould Physarum polycephalum is a large aggregate of protoplasm and behaves like an amoeboid cell, exhibiting rhythmic contraction everywhere within the organism. Phase dynamics of these oscillations were studied in relation to the global organization of amoeboid behavior, by analysing the thickness oscillation, isotonic tension and the motive force of the streaming. Usually the plasmodium showed synchrony, the phase of the oscillation being the same everywhere excepting the peripheral part. We found several situations where this in-phase relationship switched to anti-phase. This occurred either at the early stages of the plasmodial coalescence, or when a single plasmodium was nearly separated by partition, or when the streaming of the protoplasm was hindered by applying the hydrostatic pressure. Furthermore, the motive force of the protoplasmic streaming increased once the anti-phase relationship was established. In this way, the weak interactions among plasmodial parts induce the switching of phase relationship from in-phase to anti-phase, and this transition in turn acts to increase the interaction by promoting a rapid mixing of the protoplasm. This global feedback mechanism by phase switching should help maintain a large single plasmodium without separating into parts. The possible mechanism of phase switching is discussed in terms of coupled nonlinear oscillators.

Animals↗
PubMed1988

Role of astroglial cell clones in the survival and differentiation of cerebellar embryonic neurons.

To investigate the role of astrocytes in the survival and differentiation of cerebellar neurons during development, we have used astroglial cell clones, derived from 8-day postnatal cerebellar explants and which might be the in vitro equivalents of the 3 main types of cerebellar astrocytes, the Golgi epithelial cells and their Bergmann processes, the velate protoplasmic and the fibrous astrocytes (F. Alliot and B. Pessac, Brain Res., 306 (1984) 283-291). Nearly all single cells, dissociated from 15-day embryonic mouse cerebella and seeded at low density, adhered to layers of each of the cerebellar astroglial cell clones as well as to other glial lines or artificial substrates. However, the cerebellar embryonic neurons survived well only on monolayers of either the 'Golgi-Bergmann'-like or the 'velate protoplasmic'-like clones. On these layers, 60-80% of the neurons were still present after 5 days of co-culture, while only less than 5% survived on the other types of substrates. The differentiation pattern of the neurons surviving on the 'Golgi-Bergmann' and the 'velate protoplasmic' astroglial clones was studied with markers of postmitotic granule cells, the major neuronal population in adult cerebellum. The velate protoplasmic-like clone was the only one able to support the coordinate acquisition by most surviving neurons of the phenotypic characteristics of granule cells, i.e. a distinct morphology, a specific epitope binding the monoclonal antibody 7-8 D2 and immunoreactivity to glutamate. These data show a broad heterogeneity in the capacity of astroglial cell clones to support embryonic cerebellar neurons. In addition, they indicate that neuronal survival per se is not sufficient for the acquisition of a differentiated neuronal phenotype.

Animals↗
PubMed1985

A demonstration of glial filament distribution in astrocytes isolated from rat cerebral cortex.

Astrocytes have been classically described based on morphological characteristics as either protoplasmic or fibrous. However, the appearance of astrocytes at the light microscopic level following immunoreaction with glial filament acidic protein antisera or Cajal's gold-chloride method is markedly different from the morphology of Golgi-impregnated or horseradish peroxidase-filled astrocytes. The present study combines immunohistochemistry using glial fibrillary acidic protein antisera and a cellular suspension of structurally intact astrocytes isolated from the rat cerebral cortex to demonstrate that the distribution of glial filaments is limited to major astrocytic processes. Because the sheet-like processes which decorate the major branches of protoplasmic astrocytes do not contain glial filaments, the entire protoplasmic astrocyte is not visible when viewed in situ in the gray matter of the cerebral cortex. Thus staining techniques that have a propensity for the glial filaments such as antisera to glial fibrillary acidic protein and the gold-chloride method provide only a skeletal outline of the major processes of the protoplasmic astrocyte. On the other hand, fibrous astrocytes which do not have feathery decorations on major processes are visible in the cerebral cortex in their structural entirety. The present investigation also offers additional evidence of the specificity of glial fibrillary acidic protein antisera for astrocytes.

Animals↗
PubMed2006

Progress in plasmodial differentiation improves regularity of oscillating contractions in Physarum polycephalum.

Based on the knowledge about subcellular morphogenetic processes in the acellular slime mold Physarum polycephalum, we hypothesized that during differentiation of undifferentiated endoplasm to the highly differentiated complex structure of the contractile apparatus of this organism, the regularity of oscillating contractions must improve. We measured the endogenous contraction automaticity starting from the de novo generation within minutes after sampling small portions of undifferentiated endoplasm. The standard deviation of the normalized period duration of these samples was compared to the respective values of radial contractions of differentiated protoplasmic plasmodial strands. The mean normalized standard deviation in endoplasmic drops was 28.3+/-12.2%. Respective values in protoplasmic strands were 10.0+/-3.7%. The difference between the experimental groups was highly significant (p<<0.0001). We interpret the verification of our hypothesis as an indication that the very regular oscillating contractions in fully differentiated stages of Physarum require the complex structure of the sophisticated contractile apparatus, represented by the circular plasmalemma invagination system of protoplasmic strands, while the regularity is lower in stages, where the differentiation is still in progress. We believe that this is due to deficits in coordination capabilities, which need a directional and spatially oriented protoplasmic streaming as a precondition.

Animals↗
PubMed1952

The mechanism of accumulation in living cells.

When a compound enters a living cell until its activity becomes greater inside than outside, it may be said to accumulate. Since it moves from a region where its activity is relatively low to a region where its activity is relatively high, it is evident that work must be done to bring this about. The following explanation is suggested to account for accumulation. The protoplasmic surface is covered with a non-aqueous layer which is permeable to molecules but almost impermeable to ions. Hence free ions cannot enter except in very small numbers. The experiments indicate that ions combine at the outer surface with organic molecules (carrier molecules) and are thus able to enter freely. If upon reaching the aqueous protoplasm these molecules are decomposed or altered so as to set the ions free, the ions must be trapped since they cannot pass out except in very small numbers. If we adopt this point of view we can suggest answers to some important questions. Among these are the following: 1. Why accumulation is confined to electrolytes. This is evident since only ions will be trapped. 2. Why ions appear to penetrate against a gradient. Actually there is no such penetration since the ions enter in combination with molecules. The energy needed to raise the activity of entering compounds is furnished by the reactions involved in the process of accumulation. 3. Why, in absence of injury, ions do not come out when the cell is placed in distilled water. Presumably the outgoing ions will combine at the outer surface with carrier molecules and then move inward in the same way as ions coming from without. 4. Why the relative rate of penetration falls off as the external concentration increases. This is because the entrance of ions is limited by the number of carrier molecules but no such limitation exists when ions move outward since they can do so without combining with carrier molecules. 5. Why accumulation is promoted by constructive metabolism which is needed to build up the organic molecules and by destructive metabolism which brings about their decomposition. 6. Why measuring the mobilities of ions in the outer protoplasmic surface does not enable us to predict the relative rate of entrance of ions. We find for example in Nitella that K(+) has a much higher mobility than Na(+) but the accumulation of these ions does not differ greatly. This is to be expected if they enter by combining with molecules at the surface. Only if K(+) is able to combine preferentially will it accumulate preferentially. 7. Why ions may come out in anoxia and at low temperatures. If these conditions depress the formation of carrier molecules and their decomposition in the protoplasm, the balance between intake and outgo of ions will be disturbed and relatively more may come out. 8. Why the excess of internal over external osmotic pressure is less in sea water than in fresh water. As the external concentration of ions increases the rate of intake does not increase in direct proportion since the number of carrier molecules does not increase and this slows down the relative rate of intake of ions. But it does not slow down the rate of exit of ions since they need not combine with carrier molecules in order to pass out. Hence the excess of ions inside will be relatively less as the concentration of external ions increases. 9. How water is pumped from solutions of higher to solutions of lower osmotic pressure. If metabolism and consequently accumulation is higher at one end of a cell than at the other, the internal osmotic pressure will be higher at the more active end and this makes it possible for the cell to pump water from solutions of higher osmotic pressure at the more active end to solutions of lower osmotic pressure at the less active, as shown experimentally for Nitella. This might help to explain the action of kidney cells and the production of root pressure in plants.

Cells↗
PubMed2004

Effect of heat shock on the chilling sensitivity of trichomes and petioles of African violet (Saintpaulia ionantha).

Chilling at 6 degrees C caused an immediate cessation of protoplasmic streaming in trichomes from African violets (Saintpaulia ionantha), and a slower aggregation of chloroplasts in the cells. Streaming slowly recovered upon warming to 20 degrees C, reaching fairly stable rates after 4, 15, 25 and 35 min for tissue chilled for 2 min and for 2, 14 and 24 h, respectively. The rate of ion leakage from excised petioles into an isotonic 0.2 M mannitol solution increased after 12 h of chilling and reached a maximum after 3 days of chilling. A heat shock at 45 degrees C for 6 min reduced chilling-induced rates of ion leakage from excised 1-cm petiole segments by over 50%, namely to levels near that from non-chilled control tissue. Heat-shock treatments themselves had no effect on the rate of ion leakage from non-chilled petiole segments. Protoplasmic streaming was stopped by 1 min of heat shock at 45 degrees C, but slowly recovered to normal levels after about 30 min Chloroplasts aggregation was prevented by a 1 or 2 min 45 degrees C heat-shock treatment administered 1.5 h before chilling, but heat-shock treatments up to 6 min only slightly delayed the reduction in protoplasmic streaming caused by chilling. Tradescantia virginiana did not exhibit symptoms associated with chilling injury in sensitive species (i.e. cessation of protoplasmic streaming in stamen hairs and increased ion leakage from leaf tissue).

Journal Article↗
PubMed1964

ORIGIN AND PROPERTIES OF NATURALLY OCCURRING HAPTEN FROM ESCHERICHIA COLI.

Anacker, R. L. (National Institute of Allergy and Infectious Diseases, Rocky Mountain Laboratory, Hamilton, Mont.), R. A. Finkelstein, W. T. Haskins, M. Landy, K. C. Milner, E. Ribi, and P. W. Stashak. Origin and properties of naturally occurring hapten from Escherichia coli. J. Bacteriol. 88:1705-1720. 1964.-Haptens found in preparations of endotoxin and in fractions of disrupted cells, particularly one termed "native hapten," which appeared to be associated with the protoplasm of cells rather than with cell walls, have been further investigated with a view to establishing their origin and composition as well as their host-reactive properties. For this purpose, cells from a smooth strain of Escherichia coli O111:B4 were either extracted directly or disrupted and separated into cell-wall and protoplasmic fractions. Haptens were obtained by gel filtration of endotoxins, by trichloroacetic acid extraction of protoplasm, and by a mild acid hydrolysis of endotoxin. Several lines of evidence indicated that native hapten originated in the protoplasm rather than by autolysis or degradation of cell-wall endotoxin during procedures employed in disruption. In gel diffusion and quantitative precipitin tests, no hapten was identical with endotoxin, but native hapten was serologically the most complex of the haptens and precipitated the most antibody. Native and acid haptens, on a weight basis, fixed about 1% of the quantity of complement fixed by homologous endotoxin. Haptens did not stimulate the production of antibodies in mice or rabbits and did not elicit endotoxic host reactions. Chemically, native hapten differed from endotoxin and from acid hapten in that it lacked phosphorus, heptose, long-chain fatty acids, and 2-keto-3-deoxyoctonate. These substances did not appear to be determinants of antigenic specificity, but they may provide necessary bonds for assembling hapten-like units into fully antigenic and toxic macromolecules.

Animals↗
PubMed1979

Intracellular marking with lucifer yellow CH and horseradish peroxidase of cells electrophysiologically characterized as glia in the cerebral cortex of the cat.

Intracellular microelectrodes filled with either Lucifer Yellow CH, a highly florescent dye, or horseradish peroxidase (HRP) were used to electrophysiologically characterize and mark cells in the cerebral cortex of cat. Fifty-eight cells, characterized electrophysiologically as glia, were marked with Lucifer Yellow CH. All were identified as protoplasmic astrocytes, and included cells in the glia limitans of the molecular layer. An additional 54 cells, similarly characterized as glia, were labeled with HRP. The results were the same; only protoplasmic astrocytes were labeled. The "staining quality" of the glia labeled with HRP was superior to that of cells injected with Lucifer Yellow; greater lengths of individual processes were revealed, and they could often be followed to blood vessels where they ended on the walls of vessels with expanded perivascular end-feet. The observations indicate that the many previously reported studies on presumed glial cells in the cat cerebral cortex have characterized the behavior of protoplasmic astrocytes. Neurons were also marked during these experiments. The "staining" quality of the Lucifer Yellow filled neurons was excellent; dendritic spines, axons, and axon collaterals were clearly visible. These fine neuronal details were not as well revealed after HRP labeling. High resting membrane potentials (RMP's) were not a prerequisite for obtaining well-marked neurons (mean RMP of Lucifer Yellow filled neurons was -33.6 mV; mean RMP of HRP filled neurons was 42.3 mV). In contrast, the mean RMPs of Lucifer Yellow and HRP marked glia was -68 Mv and -75 mV respectively, and the quality of "staining" appeared to be more closely related to the RMP.

Animals↗
PubMed1998

Effect of cryopreservation on the fine structure of spermatozoa of rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta F. fario)

Freeze-fracture electron microscopy has revealed that changes are induced in the organization of the plasma membranes of spermatozoa of rainbow and brown trout when they are cryopreserved. Electron-micrographic images of spermatozoa that had not been exposed to cryopreservation showed a protoplasmic surface with particles homogeneously distributed. The concentration of the particles was low. In the median portion of the tail the authors observed longitudinal strips, consisting of particles located along the bands. In the neck of the spermatozoon the particles were aggregated in a chaotic manner. Electron micrographs of trout spermatozoa that had been cryopreserved showed particles grouped in rounded clusters on the protoplasmic surfaces of both the head and the tail. In some spermatozoa, folding of the protoplasmic membrane, with the particle-free sites, were found. The dimensions of the clusters and the spaces that were free of particles suggest that, after thawing, the spermatozoa of the brown trout is likely to experience greater difficulty in restoring their physiological integrity than those of the rainbow trout. These results suggest that the membrane proteins of spermatozoa of the species of trout that were studied possess high motility and diffuse, with the formation of clusters, in very short periods of time (about 30 s). The changes in membrane structure of the trout spermatozoa following cryopreservation appear to indicate high lability. Copyright 1998 Academic Press.

Journal Article↗
PubMed1988

Historical perspective: the problem of the origin of life in the context of developments in biology.

The structure of the history of scientific ideas on the origin of life, after Darwin's theory of evolution brought the problem into focus, is discussed. 19th-century theories in the mainstream of historical development already included some notion of chemical evolution. These theories were limited, however, by their reliance on a protoplasmic view of life, according to which the protoplasmic substance combines all vital properties. It was only when this holistic concept of protoplasm was abandoned that a clear distinction between different vital functions such as metabolism and replication was made. This led to two schools of thought in the origin of life field, one inspired by biochemistry and one by genetics. Oparin's theory, which was rooted in the metabolic traditions of biochemistry, provided a model which has had a lasting impact in methodological terms and which helped to transform the field from a largely theoretical one to an area of active research. Genetically based theories, on the other hand, had a delayed impact in this respect, because of long-lasting uncertainty regarding the structural basis of gene function.

Genes↗
PubMed1977

Rearrangement of intramembranous particles and fusion promoted in chicken erythrocytes by intracellular Ca2+.

Ca2+ was introduced into fresh and ATP-depleted chicken erythrocytes through the aid of the inophore A23187. Intracellular Ca2+ (10-40 mM) induced fusion in ATP-depleted cells after 30-60 min incubation at 37 degrees C, but not in fresh cells. Fresh cells underwent a higher degree of haemolysis than ATP-depleted cells after accumulation of Ca2+. Uptake of Ca2+ was the same in these two systems. Intracellular Ca2+ induced rearrangement of intramembranous particles, as revealed by freeze-etching studies. The intramembranous particles in the protoplasmic face of fractured membranes obtained from fresh cells incubated with 1 mM of Ca2+ were more scattered and their density was lower than in control cells. Incubation with higher concentrations of Ca2+ (10-40 mM) induced transient changes in the intramembranous particles' density with the appearance of protrusions and depressions on the protoplasmic and exoplasmic faces of the fractured membranes, respectively. These effects were reversible upon removal of Ca2+ by washing the cells with ethyleneglycol bis(alpha-aminoethylether)-N,N'-tetraacetic acid; rearrangement of intramembranous particles was less evident after accumulation of Ca2+ in ATP-depleted cells, whose fractured membranes did not contain any protrusions or depressions. Transferring Ca2+-loaded cells to the cold caused the formation of large smooth areas devoid of intramembranous particles in the protoplasmic face of the fractured membranes. Cells containing Ca2+ appeared spherical, and removal of Ca2+ restored the normal oval shape of chicken erythrocytes.

Adenosine Triphosphate↗
PubMed1983

Numerical densities of intramembrane particles in the cardiac sarcolemma of normal and myopathic Syrian hamsters.

The purpose of this study was to examine the density of intramembrane particles in the cardiac sarcolemma of normal and myopathic Syrian hamsters in a search for a morphological marker of a putative membrane defect. Hereditary cardiomyopathy in hamsters is manifested by progressive multifocal skeletal and cardiac muscle necrosis. Although pharmacological and biochemical data suggest a molecular defect in the cardiac sarcolemma, there has been no morphological observation that would substantiate this suggestion. We quantified numerical densities of intramembrane particles in freeze-fractured sarcolemmal membrane protoplasmic and extracellular faces from the left and right ventricular myocardium of female 25-day-old hamsters of the U.M.-X 7.1 cardiomyopathic line compared with sex- and age-matched randomly bred Syrian hamsters. Intramembrane particle numerical densities significantly increased above control values in cardiomyopathic hamsters for protoplasmic face (from 2188 +/- 61 to 2454 +/- 89 microns-2 in left ventricle (P less than 0.025) and from 2255 +/- 83 to 2574 +/- 56 microns-2 in right ventricle (P less than 0.001]. There was no significant difference between intramembrane particle densities of normal and cardiomyopathic hamsters for extracellular face (707 +/- 45 and 687 +/- 49 microns-2 in the left ventricle and 742 +/- 41 and 746 +/- 28 microns-2 in the right ventricle). These results implicate an increase of protoplasmic face-associated integral proteins in the sarcolemmal membrane of cardiomyopathic hamster. The significance of this finding is not known. It may represent an adaptive change related to a molecular defect in the sarcolemmal membrane of cardiomyopathic hamster.

Animals↗
PubMed1984

Infection of rat brain primary cell cultures with an avirulent A7 strain of Semliki Forest virus.

The ability of A7 Semliki Forest Virus (SFV) to infect primary brain cell cultures has been examined using cultures prepared from 1-2-day neonatal rat cerebral hemispheres. These cultures, characterised immunocytochemically using cell-specified markers, contain mainly GFAP+ protoplasmic astrocytes and smaller multiprocessed A2B5+ cells, probably fibrous astrocytes. 10% of the cells are GC+ oligodendrocytes and some neurones are also present. These cultures support virus growth and a cytopathic effect was observed. Using double labelling techniques with the cell-specific markers and anti-SFV antibody A7 has been shown to readily infect cells which carry either the A2B5+ antigen or galactocerebroside marker. Protoplasmic astrocytes (GFAP+/A2B5-) are not readily infected under the conditions used. The protein labelling studies using [35S]methionine show that host cell protein synthesis is not completely shut off and continues in the astrocyte protein region. These results suggest that cells derived from a common A2B5+, GFAP-, GC- progenitor glial cell, i.e. GC+ oligodendrocytes and A2B5+/GFAP+ fibrous astrocytes, are more readily infected than other brain cell types including the protoplasmic astrocytes.

Animals↗
PubMed1989

Immunocytochemical localization of nuclear 3,5,3'-triiodothyronine (L-T3) receptors in astrocyte cultures.

By means of a monoclonal antibody (mab) against the rat liver nuclear L-T3 receptor (NT3R) and a polyclonal anti-GFAp serum, it has been possible to demonstrate nuclear thyroid hormone receptors in astrocyte cultures. On day 3, 47% of GFAp+ cell nuclei were labeled by 2B3 mab. Between day 3 and day 15, the number of GFA+ cell nuclei stained by 2B3 mab increased from 47 to 75%. Thyroid hormone nuclear receptors were present in fibrous and protoplasmic astrocytes. However, they developed asynchronously in both types of astrocytes. Indeed, 60% of fibrous astrocytes were stained by 2B3 mab on day 3 and this percentage reached 77% after 8 days in vitro. In contrast, only 30% of protoplasmic astrocytes were immunoreactive for 2B3 mab on day 3 and this percentage increased slowly reaching 47% on day 8 and around 75-80% on day 15. By immunoblotting, the monoclonal antibody recognized two bands of proteins with a molecular weight of 57 and 45 kDa respectively. These proteins have the same electrophoretic mobility as [125I]bromoacetyl-LT3 rat liver nuclear L-T3 receptor. This paper presents the first immunocytochemical localization of nuclear L-T3 receptors in astrocyte cultures. Furthermore, we show that thyroid hormone receptors develop more rapidly in fibrous than in protoplasmic astrocytes.

Animals↗