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Phagokinetic tracks of 3T3 cells: parallels between the orientation of track segments and of cellular structures which contain actin or tubulin.

Phagokinetic tracks were used to determine the current direction of migration in 3T3 cells. Comparing this direction with the orientation of actin or tubulin-containing cellular structures by indirect immunofluorescence, the following results were obtained. First, the main actin-containing bundles were located at the bottom and tail end of 3T3 cells and ran parallel to the current or preceding direction of migration. Second, the 3 micrometer long rod-like structure (primary cilium), which contains tubulin and which has been observed by other investigators in transmission electron microscopy (Barnes, 1961; Sorokin, 1962; Wheatley, 1969) and in indirect immunofluorescence (Osborn and Weber, 1976), was oriented predominantly parallel to the substrate and to the current movement direction. It seems possible that the primary cilium has a role in the directional control of a migrating 3T3 cell, and that the main actin containing bundles act as substrate-attached rails along which the nucleus and bulk cytoplasm slide during displacement of the cells.

Actins↗

Cellular structures involved in albumin interaction with preadipocytes in various stages of adipogenesis, in vitro.

Albumin is the major carrier of long chain free fatty acids, that are known to be heavily utilized by fat cells. To investigate the cellular structures involved in the interaction of albumin with preadipocytes, precursor fat cells isolated from rat epididymal fat pads were cultured in Dulbecco's modified Eagle's medium supplemented with 5% fetal calf serum and antibiotics. Cell differentiation was induced by 1.7 nM insulin added to the culture medium. The ensuing cells in various stages of differentiation were incubated with albumin, alone or bearing oleic acid, adsorbed to 5 nm gold particles (Alb-Au or Alb-OA-Au) for 10 min at 37 degrees C. As control, polyethyleneglycol-gold and IgG-gold complexes were used in same conditions. After extensive washing, the cell monolayer was fixed in 2.5% glutaraldehyde, and processed for electron microscopy. Examination of thin sections and morphometric analysis showed that: a) in the process of adipogenesis induced in vitro four cell types were simultaneously present: fibroblast-like cells (F), early adipocytes (EA), adipocytes (A) and aged adipocytes (AA); b) upon incubation with Alb-Au, the probe labelled preferentially coated pits and coated vesicles and only some uncoated vesicles; in time the tracer was found in endosomes, occasionally, multivesicular bodies and lysosome-like structures. The binding and uptake was dependent on the stage of cell differentiation, being less pronounced in fibroblast-like cells and increased in later stages (A and AA); c) Alb-OA-Au labelled the same structural features like the Alb-Au (i.e. coated pits and vesicles); however, the binding to the cell membrane was fourfold higher than that of Alb-Au in F and EA stages and was maintained at the same level in A and AA stages; d) control probes were taken up only by adipocytes and aged adipocytes, cells that express a strong phagocytic function for all tracers used. These results indicated that albumin bound to gold and especially albumin-bearing fatty acids-gold, bind preferentially to coated pits and vesicles of preadipocytes. Further internalization of albumin-gold particles seems to be non-specific, since both the probes and the controls adsorbed to gold were delivered to the same endosomal-lysosomal compartment.

Adipocytes↗

[Enhancing the reparative potentials of the cellular structures in the hypothalamus by the preliminary adaptation of animals to hypoxia and stress].

Effects of preliminary adaptation of rats to hypoxic hypoxia and short-term stress environments on the dynamics of the repair changes of RNA and proteins in the cellular structures of supraoptic (SON) and paraventricular (PVN) nuclei of hypothalamus are studied. Adaptation to hypoxia and short-term stresses is found to increase the potential of protein-synthetizing system of neuro- and gliocytes of SON and PVN. It prevented a marked decrease in the RNA and proteins contents during a post-stress period and increased an intensity of the repair processes in the nuclei being tested. Experimental findings are indicative of a heterochronic pattern of repair processes in hypothalamic neurosecretory structures which to a great extent can be related with their potentials to rearrange the intracellular metabolism for maximum functional maintenance during stresses.

Adaptation, Physiological↗

Ontogenic activities and interactions of the lactate dehydrogenase isozymes with cellular structure in the guinea pig.

The LDH activities and isozyme distributions associated with soluble and particulate fractions of five major tissues have been followed during the development of the guinea pig. Evidence has been provided for an appreciable degree of interaction between LDH and cellular structure in all these tissues (liver, kidney, skeletal muscle, brain and heart) at all stages of development, but particularly in the early foetal stages. These data have been discussed in relation to the nature and extent of this binding and the correlations with the metabolic emphases in these tissue situations during development.

Animals↗

Heat-resistant variants of the Chinese hamster ovary cell: alteration of cellular structure and expression of vimentin.

Three heat-resistant mutant cell lines (78-1, 78-2, 78-3) were previously selected from Chinese hamster ovary cells. In this study, we investigated whether the differences in intrinsic thermal sensitivity result from alteration of stress protein levels or cellular structural changes. Although there was no significant difference in the levels of stress proteins, i.e., constitutive HSP70 in wild type and three heat-resistant mutant strains, there were marked differences in the amounts of vimentin among the cell lines. Two-dimensional gel electrophoresis and Western blot showed a 2.3-2.9-fold increase in the level of vimentin in the mutant cells under normal growth conditions. Northern blot also revealed higher amounts of vimentin mRNA in the mutant cells. Electron microscopy and immunofluorescence suggest that increased amounts of the vimentin-containing intermediate filaments are correlated with the heat-resistant phenotypes.

Animals↗

[Low-molecular RNA subcellular localization and its binding strength with cellular structures].

Some fractions of low molecular weight (LMW) nuclear RNAs were shown to be present in the cytoplasm of rat liver cells. In addition to known 4S tRNA, 5S and 5,8S rRNAs U3 and 8S1 LMW nuclear RNAs, 8SII and 8SIII LMW RNAs have been detected in RNA preparations of free total and membrane-bound polysomes. The U3 and 8SI polysoma I RNAs seem to be associated with high molecular weight polysomal RNA. Using thermal phenol fractionation, that some LMW RNAs were shown to be slightly bound to the cellular structures whereas some others are bound more tightly. Considerable amounts of LMW RNAs are tightly bound to the chromosome-nucleolar apparatus. They can be extracted only at 85 degrees C. The data presented are discussed with regard to LMW nuclear and polysomal RNAs functions.

Animals↗

Collateral branch formation related to cellular structures in the axon tract during corticopontine target recognition.

The corticopontine projection develops exclusively by collateral branches that form along the length of corticospinal axons days after they have passed their hindbrain target, the basilar pons. In vitro evidence suggests that the basilar pons releases a diffusible activity that initiates and directs the growth of collateral branches. This study investigates whether contact-dependent mechanisms may also influence the formation of collateral branches. By using immunocytochemistry, electron microscopy, and neuronal tracing techniques, we examined the region of the axon tract, the cerebral peduncle, overlying the basilar pons for cellular structures that correlate spatially and temporally with collateral branch formation. We found that radial glia are excluded from the tract. Oligodendrocyte precursors are found only at low density. Although mature astrocytes are absent, immature astrocytes are present throughout the tract. However, our evidence does not suggest a direct role for glial cell types in collateral branch formation. In contrast, dendrites of basilar pontine neurons are transiently present in the tract during the time of collateral branch formation. Although collateral branches are observed in regions of the tract devoid of dendrites, the orientation and location of most collateral branches correlates at the light microscopic level with dendrites. Electron microscopy reveals sites of increased collateral branch formation near neuronal cell bodies or dendrites. However, cell processes, whether dendritic or otherwise, are rarely found in direct contact with collateral branch points. A common and unexpected feature is the bundles of corticopontine collateral branches, oriented transversely to their parent corticospinal axons and directed across the tract to the basilar pons. Dendrites were often apposed to or embedded within the transverse bundles. These findings suggest that dendrites are not essential for collateral branch formation but that they may enhance this process and define discrete preferred locations for collateral branch initiation and elongation within the cerebral peduncle.

Animals↗

[Effects of electric fields on the cellular structure of the regenerated nerve].

Eighty adult SD rats were randomly divided into four groups of 20 each: 1. local electrostimulation of nerve stump (Group LS); 2. electrostimulation of myeloneure (Group N); 3. electrostimulation of the denervated muscle (Group M); and 4. Control (Group C). 5 millimeters of the left sciatic nerve of the rat was excised and the severed nerve was bridged with a silicon tube. The gap between the stumps was about 10 millimeters. On the 3rd day and the 1st, 2nd, 3rd, 4th week after surgery, samples were taken and processed for microscopic examinations to observe the effects of electric fields on cellular structural development of regenerated axons, especially on the proliferation and migration of Schwann's cell and fibroblasts, etc. The number of the axon and capillary, the degree of axon maturity and myelinated axon proportion were also observed. The results showed that electrostimulation enhanced nerve regeneration by, 1. promoting the form of matrix bridge; 2. stimulating Schwann's cell and fibroblast migration and proliferation; 3. promoting vascularization of regenerating nerves; and 4. direct electrophoretic effects on structural protein of growth cone.

Animals↗

Cellular structure and function of rat fat cells in the primary culture.

A primary culture of undigested fat tissue fragments was used to obtain fat cells in vitro. On day 2 of culture, immature fat cells, which are fibroblast-like fat cells containing fine lipid droplets, appeared, surrounding the fat tissue fragments, and began to proliferate extensively. Afterwards, these fibroblast-like fat cells grew to become multilocular fat cells containing larger intracytoplasmic lipid droplets, and differentiated further into unilocular fat cells containing a single large intracytoplasmic lipid droplet. Treatment with dibutyryl-cAMP, which is a second messenger of the lipolytic factor, caused the cultured fat cells to retract, and the intracytoplasmic lipid droplets of those fat cells became finely granulated and decreased along with an increase of hormone-sensitive lipase activities. Conversely, administration of insulin caused the lipid droplets in the fat cells to increase and become larger along with an increase of alpha-glycerophosphate dehydrogenase activities. These findings indicate the occurrence of lipolysis and lipogenesis of fat cells in vitro. Immuno-cytochemistry revealed that vimentin surrounded intracytoplasmic lipid droplets, and became distinct with an increase of lipid droplets through lipogenesis in the fat cells. Vimentin seems to be correlated to the behavior of lipid droplets in the fat cells. Fat cells in this study showed the appropriate cellular structures and functions in response to stimulation of lipolysis and lipogenesis under culture conditions. It is expected that in vitro culture of fat cells will facilitate cell biological elucidation of obesity in the future.

Adipocytes↗