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At least 397 records · Page 22Linked to original sources
PubMed1981

Acoustic trauma effects with varying exposure times.

Guinea pigs have been exposed to 20 kHz for 2 h, 1 h, 30 min, 15 min, or 7.5 min, and the resultant damage was observed 3, 6, and 12 weeks post exposure. The areas of outer hair cell damage were statistically compared. Significant differences were obtained after a 3-week survival time with the shorter exposure times, but this difference was no longer apparent after 12 weeks. Myelinated nerve fibre and inner hair cell damage occurred only with total outer hair cell loss.

Animals↗
PubMed1983

Redistribution of microfilament-associated proteins during the formation of focal contacts and adhesions in chick fibroblasts.

The roles of the microfilament-associated proteins vinculin, alpha-actinin, myosin and filamin have been studied by immunofluorescence and double fluorescence in conjunction with interference reflection microscopy (IRM), during the development of focal contacts and focal adhesions in a chick fibroblast system which initially has no such adhesion specializations but then develops them sequentially over a 48 h period. Without exception, all focal contacts and focal adhesions contain both vinculin and alpha-actinin at every stage that we can detect by IRM or by double staining to reveal the associated microfilament bundles. Indeed the appearance of small bodies containing alpha-actinin and vinculin is shown to precede focal contact formation in our model system and such structures (not visible by IRM) are proposed to be the precursors of focal contacts and adhesions. Myosin and filamin are distributed generally with some reticular patterning in the early motile cells which lack the focal specializations, but as focal contacts and adhesions form these proteins become progressively recruited into the associated microfilament bundles. Only then do we see the marked depletion that has been reported earlier of diffusely distributed myosin and filamin in the leading lamella. Although this is not initially associated with any change in the motile status of the cells, the recruitment of these microfilament-associated proteins into stress fibres is proposed to occur in preparation for anchorage and bracing of cells to the substratum when they later become stationary.

Actinin↗
PubMed1978

[Primary shape change of platelets in vitro (author's transl)].

Studies with interference contrast microscopy reveal that platelets undergo a typical shape change within 30--60' after venepuncture, i.e. swelling, formation of large tentacles, tiny protrusions and vesicles at the platelet surface. This "shape change" can be observed in citrated blood and PRP, heparinized blood and EDTA-blood as well. It is enhanced by low incubation temperatures (4 degrees C, 10 degrees C) and delayed at 37 degrees C as compared with room temperature. An increased number of primarily shape changed platelets is found if platelets are strongly mechanically irritated at blood sampling. The shape change is partly reversible in vitro, it is completely or almost completely reversible in vivo. Some antiaggregating agents inhibit the in vitro shape change at varying degrees (Bencyclan, SH 869 greater than ASA greater than D-Propranolol). The shape change is partly inhibited after oral or i.v. administration of ASA. A typical transformation of platelets into "spheric" forms can be observed following the addition of Bencyclan, SH 869 and D-Propranolol to PRP in vitro. The spontaneous "primary shape change" which occurs in PRP or blood after blood sampling is probably different from the secondary ADP-induced shape change. The primary shape change may influence the results of different platelet function and aggregating tests. The shape change kinetics of "healthy" subjects and patients with Hodgkin's disease differ significantly. The described method may gain more clinical interest in the future.

Aspirin↗
PubMed1984

Astrocyte cell lineage. IV. Changes in the organization of microfilaments and adhesion patterns during astrocyte differentiation in culture.

The organization of microfilaments using NBD-phallacidin and cell adhesion to substratum by surface reflection interference microscopy was examined during differentiation of astrocytes in colony cultures and correlated with motile behaviour of cells. Disaggregated cells from neopallium of 12-day-old or newborn DBA/1J mouse embryos were used to establish colonies and astrocyte precursor cells at various stages of differentiation along the astrocyte lineage were examined after 3 days, 1, 2 and 4 weeks in culture. The earliest astrocyte precursor cells, the glioblasts, are stationary and form epithelial-type colonies which adhere to the substratum primarily around the edge where large microfilament bundles are found. Bundles of microfilaments are also present around the apical ends of closely packed cells. As the epithelial cells start to separate and transform into flat proastroblasts, adherens-type junctions which have a zig-zag appearance and are associated with microfilaments form between adjacent cells. In the highly motile astroblasts these junctional regions break down into multiple smaller regions where the separated cells remain in contact through fine processes. The astroblasts also have stress fibres, focal contacts with substratum, foci from which microfilament bundles radiate and a complex pattern of fine, circumferentially oriented bundles of microfilaments. This elaborate organization of microfilaments disappears as the motile astroblasts differentiate into stationary fibrous astrocytes that have little polymerized actin and lack focal contacts. These results show that stationary astrocyte precursor cells in vitro go through a highly motile stage having a characteristic distribution of microfilaments and focal contacts before becoming stationary again. We consider that the motile stage could correspond to the stage in vivo when astrocyte precursor cells migrate from the ventricular and subventricular regions to take up position in different parts of the developing brain.

Actins↗
PubMed1996

Actin and the coordination of protrusion, attachment and retraction in cell crawling.

To crawl over a substrate a cell must first protrude in front, establish new attachments to the substrate and then retract its rear. Protrusion and retraction utilise different subcompartments of the actin cytoskeleton and operate by different mechanisms, one involving actin polymerization and the other myosin-based contraction. Using as examples the rapidly locomoting keratocyte and the slowly moving fibroblast we illustrate how over expression of one or the other actin subcompartments leads to the observed differences in motility. We also propose, that despite these differences there is a common coordination mechanism underlying the genesis of the actin cytoskeleton that involves the nucleation of actin filaments at the protruding cell front, in the lamellipodium, and the relocation of these filaments, via polymerization and flow, to the more posterior actin filament compartments.

Actins↗
PubMed1987

Effect of oocyte quality and sperm characteristics on the number of spermatozoa bound to the zona pellucida of human oocytes inseminated in vitro.

Sperm characteristics and oocyte quality may play a role in in vitro fertilization. The objective of this paper is to analyze the effect of the quality of oocytes, the husband's semen characteristics, and category of the couple's infertility on the number of spermatozoa bound to the zona pellucida. One hundred eighty-one oocytes which failed to fertilize or failed to cleave were fixed in 2.5% glutaraldehyde 40 to 60 hr after insemination in vitro and examined under interference microscopy and the number of sperm bound to the zone pellucida was determined. The means +/- SD of sperm bound to mature, immature, and atretic oocytes were 51.0 +/- 50.7, 7.3 +/- 12.1 10.4 +/- 17.8, respectively. Fertilized mature oocytes (81.0 +/- 53.3) had a significantly higher number of sperm bound to zonae compared to unfertilized oocytes (41.8 +/- 47.3). It is concluded that the number of sperm bound to zonae is functionally important. The sperm motility and the number of motile sperm used to inseminate oocytes were significantly correlated with the number of sperm bound to zonae, whereas sperm morphology and sperm concentration did not correlate. This study supports the notion that sperm motility is the single most important factor influencing fertilization of human oocytes in vitro.

Female↗
PubMed1987

Dry mass, DNA and non-histone protein determinations in lung cancer cells.

Cell nuclei from two biopsies of bronchial mucosa, seven squamous-cell carcinomas and six small-cell carcinomas of the lung were investigated. DNA and non-histone protein (NHP) content were simultaneously determined in Feulgen-Naphthol Yellow S-stained smears by means of multiple plug cytophotometry. In addition, the nuclear dry mass of cells originating from the same populations was measured by interference microscopy. DNA histograms of carcinomas were characterized by DNA stemlines being situated in the diploid range in four out of six small-cell carcinomas and in the hyperdiploid to hypertetraploid range in six out of seven squamous-cell carcinomas. Polyploid values (up to 8 c) were seldom found in small-cell carcinomas whereas squamous-cell carcinomas showed a broader dispersion approaching the 16 c level. The similarity of the NHP distributions with the DNA histograms was more marked in small-cell carcinomas than in the squamous-cell carcinomas. In comparison with the NHP distributions of normal bronchial epithelial cells, those of carcinomas were shifted to higher values. The increased NHP content was found to be a more prominent sign of malignancy in small-cell carcinomas than the DNA mass. The increase in nuclear dry mass in carcinomas was mainly caused by the accumulation of NHP in tumour cell nuclei.

Bronchi↗
PubMed1986

Improvement in the staining and in the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level.

The argyrophilic proteins of the nucleolar organizer region (Ag-NOR proteins) were specifically localized at the optical level with a modified one-step silver technique performed at 20 degrees C. This method was applied to various materials including cells in smears, chromosomes, semi-thin sections of plastic-embedded cells and sections of paraffin-embedded human pathological tissues. In order to improve the visualization of the silver deposits we tested various modes of imaging, including bright-field, Nomarski contrast, reflected light and combined Nomarski contrast with reflected light. The use of Nomarski contrast is useful to define precisely the phases of mitosis. The use of reflected light, which is based on the ability of silver to reflect incident light specifically, gives images with an improved resolution compared to bright-field.

Cell Line↗
PubMed1978

Direct visualization of epithelial morphology in the living amphibian urinary bladder.

Differential interference-contrast microscopy has been applied to the study of amphibian urinary bladders, in vitro. It is demonstrated that well-resolved images can be obtained with little loss of tissue viability. Direct observations have been made on the structure of microvilli, the distribution of mitochondria in the mitochondria-rich cells, and the patency of lateral intercellular spaces. It is noted that the effective viscosity of cytoplasm is very high--that it is apparently a gel in which there is no Brownian movement of organelles. The frequency, shape, and pattern of distribution of granular and mitochondria-rich cells is determined for the commonly studied varieties of Bufo marinus. Bladders from Colombian toads contain more and larger mitochondria-rich cells than do those of the Dominican variety. There is no specific arrangement of cell-cell contacts to suggest a structural basis for cooperativeness of action. Finally, a longitudinal study of osmotically-induced changes in the structure of the "tight" or "limiting" junctions establishes the validity of previous findings by electron microscopy.

Animals↗
PubMed1984

Substratum attachment of embryonic mesoderm cells in culture.

The cell-substratum adhesive characteristics of cultured chick embryo primary mesoderm cells have been examined by interference reflection microscopy and transmission electron microscopy under various conditions. Correlations were drawn between the type of adhesion and the degree of motility shown by the cells. During the rapid spreading and motility of cells cultured on fibronectin-containing substrata, focal contacts (10 to 15-nm gap) were rare and close contacts (about 30-nm gap) were predominant. By contrast, when the cells were immobile, after 5 d in culture, extensive focal contacts were present, together with stress fibers. The results indicate that tight cell-substratum contact is incompatible with rapid cell motility and that fibronectin acts by inducing the formation of close contacts rather than focal contacts.

Animals↗
PubMed1998

Cytoskeleton and polyploidy after maturation and fertilization of cryopreserved germinal vesicle-stage mouse oocytes.

PURPOSE: Our purpose was to assess the effect of cryopreservation on cytoskeleton of germinal vesicle (GV) mouse oocytes and determine whether irreversible spindle damage and related digyny associated with cryopreservation of metaphase II (MII) oocytes can be avoided. METHODS: The GV oocytes were cryopreserved using a slow-cooling (0.5 degree C/min) and slow-thawing (8 degrees C/min) protocol in 1.5 M dimethylsulfoxide supplemented with 0.2 M sucrose and analyzed before and during fertilization by multiple-label fluorescence and differential interference contrast microscopy techniques. RESULTS: When examined after in vitro maturation, the vast majority (> 95%) of cryopreserved and control oocytes displayed normal microfilament and microtubule organization. With respect to barrel-shaped spindle and normal chromosome alignment, no significant differences were observed between cryopreservation (78 and 86%, respectively) and control (85 and 95%, respectively) groups. In fertilization experiments, spindle rotation, formation of the second polar body, and pronuclear migration were displayed by similar percentages of cryopreserved (96, 94, and 37%, respectively) and control (98, 97, and 45%, respectively) oocytes, indicating normal functionality of the cytoskeleton during this period. However, pronuclear formation was significantly inhibited by cryopreservation (81%) compared with controls (100%). Regarding digyny and polyspermy, no significant increase was observed after cryopreservation (3 and 10%, respectively) compared with controls (3 and 6%, respectively). CONCLUSIONS: Cryopreservation of mouse oocytes at the GV stage is particularly advantageous to circumvent the spindle damage and increased digyny noted after cryopreservation of MII oocytes.

Actin Cytoskeleton↗