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Correlation between effects of 24 different cytochalasins on cellular structures and cellular events and those on actin in vitro.

To compare the effects of cytochalasins on the cellular level with those on the molecular level, 24 cytochalasins, 20 natural compounds and 4 derivatives, were used. The following effects were tested for each of 24 cytochalasins; (a) four high dose (2-20 muM) effects on the cellular level: rounding up of fibroblastic cells, contraction of actin cables, formation of hairy filaments containing actin, and inhibition of lymphocyte capping; (b) a low dose (0.2-2 muM) effect: inhibition of membrane ruffling; and (c) two in vitro effects: an inhibition of actin filament elongation (the high affinity effect [low dose effect] in vitro) and an effect on viscosity of actin filaments(the low affinity effect [high dose effect] in vitro). These results indicated that there are almost the same hierarchic orders of relative effectiveness of different cytochalasins between low and high dose effects and between cellular and molecular effects. From the data obtained with the 24 cytochalasins, we have calculated correlation coefficients of 0.87 and 0.79 between an effect in vivo, inhibition of capping, and an effect in vitro, inhibition of actin filament elongation, as well as between inhibition of capping and another effect in vitro, effect on viscosity of actin filaments, respectively. Furthermore, a correlation coefficient between the high affinity effect and the low affinity effect determined in vitro was calculated to be 0.90 from the data obtained in this study. The strong positive correlation among low and high dose effects in vivo and those in vitro suggests that most of the effects caused by a cytochalasin, irrespective of doses or affected phenomena, might be attributed to the interaction between the drug and the common target protein, actin. In the course of the immunofluorescence microscope study on cytochalasin-treated cells using actin antibody, we have found that aspochalasin D, a 10-isopropylcytochalasin, strongly induced the formation of rodlets containing actin in the cytoplasm of the treated fibroblasts. In contrast, the other cytochalasins, including cytochalasin B, cytochalasin C, cytochalasin D, and cytochalasin H, were found to induce the formation of nuclear rodlets. Both cytoplasmic and nuclear rodlets found in the cytochalasin-treated cells were similar in ultrastructures to those induced by 5 to 10 percent (vol/vol) dimethyl sulfoxide in the same type of cells.

Actins↗

[Preparation and characterization of low protein, cell structural material. II. Materials with cellular structure from carrot and white cabbage].

The molecular parameters and functional properties of low-protein materials with a cellular structure are determined by both the conditions of preparation and the kind of raw material used. Compared to alcohol-insoluble substance, the conditions of preparation were modified. When an alkaline extraction and a favorable combination of processing steps is used, materials with special low protein and high pectin contents result. In addition, such functional properties as water binding capacity and cation exchange capacity of the materials are improved. After having prepared various materials with a cellular structure from apple, carrot, and white cabbage, it was found that the amount of solvent needed and the yield, as well as the molecular parameters and the functional properties were also determined by the kind of raw materials used. For instance, material with a cellular structure from apple is low in protein and high in water binding capacity. Whereas, materials with a cellular structure from carrot, which contains a deesterified pectin component, is characterized by a good yield, a high pectin content, and an excess of 1.5 mmol/g of free carboxyl groups.

Brassica↗

Quantitative motion analysis and visualization of cellular structures.

The availability of cellular markers tagged with the green fluorescent protein (GFP) has recently allowed a large number of cell biological studies to be carried out in live cells, thereby addressing the dynamic organization of cellular structures. Typically, microscopes capable of video recording are used to generate time-resolved data sets. Dynamic imaging data are complex and often difficult to interpret by pure visual inspection. Therefore, specialized image processing methods for object detection, motion estimation, visualization, and quantitation are required. In this review, we discuss concepts for automated analysis of multidimensional image data from live cell microscopy and their application to the dynamics of cell nuclear subcompartments.

Animals↗

Time course of atrial fibrillation-induced cellular structural remodeling in atria of the goat.

BACKGROUND: Previously we documented cellular structural changes of a non-degenerative nature in atrial myocytes after atrial fibrillation (AF) in the goat. The time course of these changes was not studied. METHODS AND RESULTS: Cellular structural changes were studied by light- and electron microscopy and immunohistochemistry in goat atria after 0-16 weeks AF. The first sign of cellular structural remodeling was a more homogeneous chromatin distribution, at 1 week of AF. Sub-structural changes in mitochondria and sarcoplasmic reticulum occurred gradually. Cellular degeneration was absent. The degree of myolysis and glycogen accumulation increased till 8 weeks of AF and did not increase further from thereon. After 16 weeks of AF, 42% of the myocytes in the right atrial free wall were affected by myolysis. The diameter of the atrial myocytes increased. Dedifferentiation of the atrial myocytes was suggested by altered expression patterns of structural proteins, such as the disappearance of cardiotin (1 week), the A-I junctional part of titin (4 weeks), desmin at the intercalated disk (ID) (8 weeks) and a gradual re-expression of alpha-smooth muscle actin. CONCLUSION: Remodeling of the cellular ultrastructure in atrial myocardium of the goat develops progressively during AF. Re-expression of fetal proteins indicate dedifferentiation of atrial myocytes, analogous to observations in hibernating myocardium of the ventricle.

Actinin↗

Ontogenic variations in the interactions of lactate dehydrogenase isozymes with cellular structure.

The interactions of the LDH isozymes with cellular structure have been studied in the major tissues of the mouse during development. Overall, there is a clear indication that interactions between LDH and cellular structure are appreciable in all major tissues and at all stages of development, and an analysis of the isozyme status of the enzyme in both soluble and bound compartments has been effected. Information has been provided supporting the preferential binding of the A type activity to actin filaments. The data provided also support the concept of pelletable B type LDH in kidney and brain during the postnatal period. Binding was particularly extensive in the early embryonic stages, and the significance of this phenomenon, and the subcellular interactions observed to varying degrees in all the tissues throughout development, have been discussed in relation to the known metabolic characteristics of these separate tissue situations.

Animals↗

Observations on cellular structures of Porphyridium cruentum.

The cellular structure of Porphyridium cruentum was studied with both light and electron microscope. The photosynthetic plastid in this red alga was found to be structurally similar to that in the Chlorophyceae and higher green plants. The phycobilins, as well as the chlorophyll, seem to be associated with the lamellae of the plastid. The pyrenoid, a region of low lamellar density, contains no tubules, and does not appear to function in synthesis or storage of reserve material. Grains of floridean starch are located in the cytoplasm, outside the plastid. Typical mitochondrial organelles were not observed. The nucleus is eccentric, and contains a nucleolus located on the inner face of the nucleus, nearest the plastid. The schedule for staining the nucleus is given in detail. Other cell structures (sheath, dictyosomes, etc.) are described. Growing cells in light of intensity leads to disruption of the parallel arrangement of the lamellar characteristic of cells grown in moderate light.

Cell Nucleus↗

Structure-activity relationship of thirty-nine cytochalasans observed in the effects on cellular structures and cellular events and on actin polymerization in vitro.

The effects of twenty-three natural cytochalasans and sixteen synthetic derivatives on actin-distribution and alteration in morphology of C3H-2K cells, inhibition of lymphocyte capping, and inhibition of actin filament elongation were compared. The effects on cellular level and the in vitro effects showed positive correlation and the structure-activity relationship observed is discussed.

Actins↗

A model study of the effects of the discrete cellular structure on electrical propagation in cardiac tissue.

The effects of the discrete cellular structure on propagation of electrical excitation in cardiac muscle were studied in a one-dimensional fiber model containing a periodic intercalated disk structure. Globally, the macroscopic velocity of propagation follows the behavior associated with propagation in a continuous tissue (except for high values of disk resistance). In addition, the computed spatial extracellular potential along the fiber is a smooth biphasic waveform and does not reflect the underlying discrete cellular structure of the tissue. Other results of the simulations demonstrate the discontinuous nature of propagation and the importance of the structure in arrhythmogenesis. Vmax displays a biphasic behavior as a function of increasing intercalated disk resistance. An initial "paradoxical" increase in Vmax (with a simultaneous decrease in conduction velocity) is followed by a decrease that leads to decremental propagation and conduction block. The time constant of the foot of the action potential (tau foot) increases monotonically with increasing intercalated disk resistance. An increase in the leakage current to extracellular space brings about a significant decrease in the action potential duration and a loss of the plateau. This major effect is accompanied by a relatively smaller decrease in conduction velocity. Collision of two activation wavefronts results in a significant (100%) increase in Vmax and a very small (0.6%) decrease in tau foot.

Action Potentials↗

Dielectrical model of cellular structures in radio frequency and microwave spectrum. Electrically interacting versus noninteracting cells.

A model of dielectrical properties of cellular structures of a tissue has been proposed. Cellular structures were presented as a composition of membrane covered spheres and cylinders that do not interact with each other. No restrictions were applied to the thickness of cellular membranes. The model was further generalized into a case of electrically interacting cells. The difference in dielectrical properties calculated with the model of electrically noninteracting versus interacting cells is inversely dependent on frequency. At biological values of cellular volume fraction near 0.7 (packed configuration) the difference is about 10%-15% in resistance and in epsilon' for frequencies near 0.1 MHz. Experimental data for myocardial tissue and theoretical data, for both interacting and noninteracting models, reasonably agree at frequencies of 1-100 MHz.

Animals↗

Circadian rhythm of the liver of male rats dosed with phenobarbital--I. Organ weight, cellular structures, glycogen contents and mitotic activity.

The circadian rhythm of the liver, namely organ weight, cellular structures (by light-microscopy), glycogen content (by periodic acid-Schiff (PAS) reaction) and mitotic activity, was studied in 166 male Sprague-Dawley rats orally treated daily at 0800-0900 with 70 (study 1) or 50 (study 2) mg/kg phenobarbital (PB) for 7 days. Thereafter, eight (study 1) or five (study 2) rats each were studied at 4-hr intervals at 1000, 1400, 1800, 2200, 0200, 0600 and 1000 through till the following day. The lighting schedule in the colony was 12:12, light:dark (light from 0600 to 1800). The liver weight was raised in PB-treated rats at all times of the day compared to controls and showed a distinct circadian rhythm with a peak at 1000 and a minimum at 2200 in PB-treated rats and the controls. The circadian rhythm of cellular structures was closely related to the hepatic glycogen content which was in good agreement with the controls, but at 1400 and 1800 the glycogen particles were more distinctly diminished in the enlarged centrilobular hepatocytes of PB-treated rats. The mitotic activity of hepatocytes was markedly increased in rats treated with PB but showed the same circadian rhythm as controls with a peak at 1000.

Animals↗

Cytoskeleton in human mammary carcinoma cells forming three-dimensional cellular structures within collagen gels.

Human mammary carcinoma cell line MCF-7 cells grown on type I collagen gels floating in a medium occasionally invaginated into the gels as a cell mass and formed cylindrical or domed structures within it. The 0.05% Triton-insoluble cytoskeleton of such cellular structures sedimented as a white flocculent layer at the boundary between 60 and 70% sucrose layers by ultracentrifugation, and consisted of 4 basal components: 54-kD (beta-tubulin), 45-kD, 42-kD (actin), and 39-kD polypeptides. By contrast, the isolated cytoskeleton of MCF-7 cells grown as monolayers on plastic substratum formed a finer cytoskeletal network with a smaller buoyant density and consisted of two distinct polypeptides with apparent molecular sizes of 80-kD and 65-kD in addition to the 4 basal components found in the morphologically developing cells. The present results indicate that the cytoskeleton of MCF-7 cells forming the three-dimensional cellular structures within collagen gels is lacking in these two polypeptides, and that it has a coarser cytoskeletal network with a greater buoyant density than that of the monolayered cells on plastic.

Actins↗

Changes in cellular structures and enzymatic activities during browning of Scots pine callus derived from mature buds.

Visible browning is a typical feature of callus cultures derived from shoot tips of mature Scots pine (Pinus sylvestris L.). Because the ability of callus to regenerate is low, we determined the effect of browning on growth and changes in cellular structure during culture. Striking alterations in cellular structure were detected by LM (light microscopy), EM (electron microscopy) and SEM (scanning electron microscopy). Accumulation of phenolic substances was shown by histochemical staining. Staining for beta-glucosidase activity of soluble proteins that had been subjected to polyacrylamide gel electrophoresis indicated lignification of cells. The measured growth rate of callus was low compared with a hypothetical growth curve. Peroxidase activity increased rapidly soon after the start of the culture period, but especially between the second and third weeks of culture. At this time, the degradation of cell membranes and browning began coincident with the loss of chlorophyll. We conclude that browning is associated with cell disorganization and eventual cell death, making tissue culture of mature pine especially difficult.

Journal Article↗

Cellular structure and function of mouse adrenocortical tumor cells Y-1 in the post-treatment state of low Ca2+.

Under lowered Ca2+ content and in the post-treatment state of low Ca2+, we studied the cellular structure and functioning in mouse adrenocortical tumor cells, Y-1. These cells had been maintained in Ham F12 medium containing 10% fetal calf serum. The Ca2+ present in this complete medium was 0.39 mM. Under a slightly lowered Ca2+ (0.29 mM) produced by EGTA, the cells had many blebs on their surfaces and specific functional activity decreased in steroidogenesis as did ACTH reactivity. In the post-treatment state of a low Ca2+, the cellular surface was covered with many short microvilli and there was greater cellular activity than in the control cells. When the Ca2+ concentration was below 0.17 mM, the cellular structure and functioning were disturbed, and there was no recovery even at the physiological Ca2+ after the removal of EGTA.

Adrenal Cortex Neoplasms↗

Circadian rhythm of liver parameters (cellular structures, mitotic activity, glycogen and lipids in liver and serum) during three consecutive cycles in phenobarbital-treated rats.

The circadian rhythm of gastric content, serum alkaline phosphatase (alk.P.), serum lipids, body weight (wt), relative (rel.) liver wt, cellular structures (by light- and electron-microscopy), mitotic activity of hepatocytes, glycogen content, protein and lipids in liver was studied in 180 male Sprague-Dawley rats orally treated at 0830-1030 with 50 mg/kg phenobarbital (PB) for 7 days. Thereafter, five PB-treated males and five controls each were studied at 4-hr intervals at 0600, 1000, 1400, 1800, 2200 and 0200 on 3 consecutive days. The lighting schedule in the colony was 12:12 = light/dark (light from 0600 to 1800). Following the rhythm of gastric emptying, the rel. liver wt showed a clear circadian rhythm with a peak at 0800. The rel. liver wt was raised in PB-treated rats at all times of the day. The circadian rhythm of cellular structures was closely related to the hepatic glycogen content which exhibited a clear rhythm with the peak also at 0800, but lowered values were found in PB-treated rats. The mitotic activity of hepatocytes was significantly increased in PB-treated rats but displayed the same circadian rhythm as controls with peaks at noon and troughs at midnight. The well-known hypertrophy of the smooth endoplasmic reticulum in PB-treated rats was not found at 0600, but was fully developed at 1400 and 2200. PB-treatment increased significantly the liver content of cholesterol, triglycerides and phospholipids. Liver cholesterol showed a clear circadian rhythm with peaks at 1800. No rhythm of liver protein, triglycerides and phospholipids was observed. In serum, levels of cholesterol were significantly elevated, those of triglycerides and alk.P. significantly lowered, while those of phospholipids were not affected by the treatment. The three serum lipids, alk.P. and beta-lipoprotein exhibited a clear circadian rhythm, while serum glucose and non-esterified fatty acids did not.

Animals↗

Influence of marihuana on cellular structures and biochemical activities.

Cannabinoids are known to affect a number of cellular systems and functions, but the basis for their action is unclear. In this paper we review the current evidence describing cannabinoid effects on various levels of cellular structure and activity and we present our current studies on the influence of delta-9-tetrahydrocannabinol, cannabidiol and cannabinol on one cellular system, the cytoskeleton. The organization of two cytoskeletal structures, microtubules and microfilaments, were examined and the mRNA levels of tubulin and actin, the major protein components of microtubules and microfilaments, respectively, were analysed.

Animals↗

[Cellular structure of propionibacteria during their multiplication].

The aim of the present study was to determine the structure of bacterial cells from Propionibacterium genus as well as their structure during the cellular division. On the basis of the observations made in the electron transmission microscope, in uranyl-acetates-tained preparations of ultra-thin specimens of bacteria, it was stated that propionic bacteria appeared in a shape of short rods, possessing regular profiles of cell walls as opposed to Gram-negative bacteria with a very creased edge line. Besides, it was observed that division of cells had place by formation of septum, most probably preceded by the division of mezosome, which is a signal for creating the divisional wall. In the conducted studies, the following phenomena were started: presence of membraneous structure of mezosomes, which is linked with the chain of circular DNA in bacterial cell, appearance of numerous ribosomes in the regions of tangled threads of nucleic acids, and existence of other undefinite elements. Mezosome present in the cell of propionic bacteria is probably linked with the cell wall at least in two places and on the surface of external cell wall at the site of its linking; it causes the change in electronic density, demonstrated by the undefined holes or scars in cell wall. This finding gives the possibility of distinguishing this genus of Propionibacterium, in the respect of morphology, from other bacteria what, in the opinion of the authors, is a new achievement in the studies on the structure of propionic bacteria.

Cell Division↗

The influence of androgens on enzymes (chymotrypsin-and trypsin-like proteases, renin, kallikrein and amylase) and on cellular structure of the mouse submaxillary gland.

1. The effect of age and androgen level on enzyme activity and cellular structure has been determined in the mouse submaxillary gland.2. A new protease which resembles chymotrypsin in its substrate specificity has been characterized in the gland.3. Activity of the chymotrypsin- and trypsin-like proteases and renin increased considerably in male mice concomitantly with proliferation of granules in the secretory tubules of the gland.4. The androgen dependence of the chymotrypsin- and trypsin-like enzymes, renin and the organelles within the secretory tubules was confirmed in castrated male mice. The activity of these enzymes increased and correlated with the appearance of intracellular granules in the secretory tubules when the castrated male mice and in addition female mice were treated with testosterone preparations.5. Kallikrein, a closely related protease, and amylase increased in activity with age but showed no sex-linked differences.6. The results suggest that kallikrein is sequestered in acinar cells whereas the androgen-dependent enzymes (chymotrypsin, trypsin and renin) are located in the secretory tubules.

Age Factors↗

The influence of calcium ions on the adsorption of glycolytic enzymes to cellular structure.

In order to provide information on the influence of Ca2+ ions on the adsorption of glycolytic enzymes to cellular structure, the release of these enzymes from digitonized cells has been studied. Increases in the calcium ion concentration were found to cause corresponding decreases in the extent of release of all the glycolytic enzymes, as well as a parallel increase in the extent of polymerization of actin. These observations have been discussed in relation to the effect of physiological concentrations of these ions on the association between glycolytic enzymes and the cytoskeleton.

Actins↗