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At least 1,225 records · Page 68Linked to original sources
PubMed1993

Fatty acid composition of lecithin is a key factor in bile metastability in supersaturated model bile systems.

We studied the effect of fatty acid saturation of biliary lecithin on bile metastability, determined by nucleation time, using model bile solutions with identical lipid compositions except for the lecithin species (total lipid concentration, 9 g/dl; cholesterol, 12 mM; lecithin, 31 mM, bile salts, 116 mM). Gel permeation chromatographic studies revealed that nonmicellar cholesterol distribution was inversely related to the degree of unsaturation of the lecithin species. Differential interference contrast microscopy and cholesterol crystal growth assay showed that a lower degree of saturation of the lecithin species was associated with a faster nucleation time and crystal growth rate. These results suggest that vesicular lecithin containing more unsaturated fatty acyl chains binds less tightly to cholesterol as compared with lecithin containing predominantly saturated fatty acids and that the biliary lecithin species modulates cholesterol crystal nucleation in bile. Also, the high ratio of cholesterol to lecithin (more than 1.0) was found in the crystal forming model biles, although the vesicle aggregation was not always observed prior to the cholesterol crystal formation. These findings indicated that there are different processes in cholesterol crystal nucleation, with or without vesicle aggregation, and that such processes depend, in part, on lecithin species in vesicles.

Bile↗
PubMed1994

Degree of fatty acyl chain unsaturation in biliary lecithin dictates cholesterol nucleation and crystal growth.

To clarify factors involved in the formation of cholesterol gallstones, we studied the relationship between the degree of fatty acyl chain unsaturation of biliary lecithin and bile metastability. We used supersaturated model bile solutions (molar taurocholate/lecithin/cholesterol ratio (73:19.5:7.5), total lipid concentration 9 g/dl) that contained equimolar egg yolk or soybean lecithins or a sn-1 palmitoyl, sn-2 linoleoyl phosphatidylcholine. Gel permeation chromatographic studies showed that the vesicular cholesterol distribution and dimension were inversely related to the degree of unsaturation of the lecithin species, estimated by reverse phase, high-performance liquid chromatography. Differential interference contrast microscopy and assay of cholesterol crystal growth showed that a higher degree of fatty acyl chain unsaturation of the lecithin species was associated with a faster nucleation time and rate of crystal growth. Our results suggest that vesicular lecithins containing more unsaturated fatty acyl chains bind less tightly to cholesterol than lecithins containing predominantly saturated fatty acids, and that the biliary lecithin species dictates, in part, the nucleation and growth of cholesterol crystals in bile.

Bile↗
PubMed1987

Absolute specificity for retrograde fast axonal transport displayed by lipid droplets originating in the axon of an identified Aplysia neuron in vitro.

Lipid droplets were found to form all along the axon of the giant cerebral neuron (GCN) of the sea hare Aplysia californica when the cell was placed in culture. The emission of yellow fluorescence by the droplets after exposure of the neuron to Nile red and their uniformly dark appearance in electron micrographs of axons fixed with glutaraldehyde and osmium tetroxide identified them as lipid droplets. In contrast to lipid droplets in fat cells and certain other cell types, these droplets were bounded by a membrane, indicating that the lipid droplet is a type of organelle that is membranated in some situations but not others. As observed by video-enhanced contrast-differential interference contrast microscopy, the droplets grew manyfold in place in the axon to diameters of 1-3 micron within 2-3 days. Often they formed coherent tandem arrays of 3-15 droplets. Droplets were usually essentially stationary but occasionally moved tens of microns by fast axonal transport, the largest spherical organelles to have been observed to undergo transport. They usually moved as singlets, sometimes as tandem arrays. The direction of transport was always retrograde (towards the cell body). Thus, an organelle need neither originate nor be modified in the axon terminal to be specified for retrograde transport. Whether or not an organelle is formed in the cell body might determine directionality. Alternatively, size might be a determining factor, with large organelles specified for retrograde transport.

Animals↗
PubMed1995

Increased collagen content of cerebral microvessels in Alzheimer's disease.

Based on previous evidence suggesting abnormalities in the brain microvasculature, we examined basement membrane collagen in isolated cerebral microvessels (CMV) from subjects with Alzheimer's disease (AD) and age-matched controls. Concentrations of hydroxyproline, the principal constituent of collagen IV, were significantly increased by 55% in CMV from AD subjects compared to controls. This result was corroborated by the finding of 60% increased total collagen content in CMV as evident by the selective binding of Sirius red dye. Hydroxyproline and collagen concentrations in samples of cerebral cortex assayed in parallel were 6-20 times smaller than in CMV and were not changed between controls and AD subjects. To further differentiate AD and control samples, fractions of CMV were solubilized and the pepsin digested collagen proteins resolved by SDS-PAGE. Upon immunoblotting, AD samples with increased collagen revealed proportionally greater specific immunoreactivities detected by antibodies to collagen IV. Our observations suggest altered collagen IV content of cerebral vessels in subjects with AD that may affect brain microvascular functions.

Aged↗
PubMed1983

Morphological changes during maturation of starfish oocytes: surface ultrastructure and cortical actin.

The cell surface and extracellular investments of oocytes of the starfish Pisaster ochraceus are analyzed by Nomarski differential interference contrast microscopy and by scanning electron microscopy. The investing coats include a thin sheet of follicle cells, a jelly coat, and a vitelline layer; their morphologies are described. Methods are outlined for systematically removing them without altering the behavior of the oocyte so that the cell surface can be examined directly. The topography of denuded oocytes changes dramatically when they are treated with the maturation-inducing hormone, 1-methyladenine. The major topographical change is the early and transient formation of prominent surface spikes. These structures arise due to the rapid, reversible polymerization of actin into stout bundles. Polymerization and subsequent depolymerization of cortical actin is monitored by epifluorescence microscopy of oocytes stained with NBD-phallacidin, a stain which is specific for polymerized actin. Based on scanning electron microscopy, spikes apparently utilize preexisting plasma membrane of microvilli, and plasma membrane is apparently lost when spikes collapse. Long after microvilli are eliminated due to spike formation, the number of microvilli is somewhat restored, especially around the animal pole where the polar body forms. A chronology of events observed during oocyte maturation is discussed with reference to the possible mechanisms and implications of polymerization and depolymerization of cortical actin.

Actins↗
PubMed1987

Changes in the phosphorylation and distribution of vinculin during nerve growth factor induced neurite outgrowth.

The mechanism of neurite initiation and elongation was studied using nerve growth factor (NGF) treatment of PC12 cells. The distribution of focal adhesion sites and of the cytoskeletal protein vinculin was determined in large, fused, multinucleated PC12 cells. In the absence of NGF, focal adhesion sites as seen by interference reflection microscopy were restricted to the cell periphery in a regular distribution. Vinculin assemblies (foci), observed by indirect immunofluorescence microscopy using affinity purified anti-vinculin antibodies, were restricted to the cell periphery at focal adhesion sites. Within 4 hr after NGF treatment of the cells, the distribution of both vinculin and focal adhesion sites began to change. Focal adhesion sites became restricted to discrete protruding portions of the cell periphery. Larger, brighter vinculin foci appeared at the tips of the cell margin extensions, concomitant with the loss of foci at locations between the protrusions. As neurites elongated focal adhesion sites and vinculin foci remained with the tips of the growth cone extensions. Both focal adhesion sites and vinculin foci were rarely seen in the perikarya of cells with elongating neurites, and these were always confined to extended portions of the cell body margin. Occasionally, vinculin foci could be seen at the proximal portion of the neurite, at bending elbows, and at discrete expansions along the length. By immunoprecipitation of vinculin from 32P-labeled cells, vinculin phosphorylation was found to be increased within 1 hr of NGF treatment. The role of vinculin phosphorylation and assembly in the formation and directional elongation of neuritic processes in response to NGF is discussed.

Animals↗
PubMed1990

Mouse oocytes regulate hyaluronic acid synthesis and mucification by FSH-stimulated cumulus cells.

Mucification (or expansion) of the cumulus cells surrounding the oocyte is thought to depend on the direct action of gonadotropins in stimulating production and deposition of hyaluronic acid (HA) in the extracellular matrix. We now report that the oocyte is essential for this process. Either follicle-stimulating hormone (FSH) at 1 micrograms/ml or dibutyryl cAMP at 2 mM induces mucification of intact cumulus cell-oocyte complexes (COCs) in vitro, but fails to stimulate mucification of isolated cumulus cells. HA synthesis by FSH-stimulated cumulus cells is only approximately 3.5% of the value achieved by FSH-stimulated COCs. Isolated oocytes cultured with or without FSH do not synthesize detectable amounts of HA but induce isolated cumulus cells to increase HA synthesis approximately 13-fold in cocultures with FSH. Medium conditioned by isolated oocytes for 5 hr induces nearly the same level of HA synthesis by cumulus cells under the same culture conditions. FSH also stimulates cumulus cells to increase synthesis of dermatan sulfate proteoglycans (DS-PGs) approximately 3-fold, but this stimulation does not depend upon the presence of oocytes. The results indicate that oocytes produce a soluble factor(s) essential in combination with FSH to stimulate HA, but not DS-PG, synthesis by cumulus cells in vitro and that this factor(s) acts independently or downstream from the FSH-induced formation of cAMP.

Animals↗
PubMed1990

U3 snRNPs and nucleolar development during oocyte maturation, fertilization and early embryogenesis in the mouse: U3 snRNA and snRNPs are not regulated coordinate with other snRNAs and snRNPs.

U3 small nuclear ribonucleic acids (snRNA) and U3 small nuclear ribonucleoprotein (snRNP), which are thought to be responsible for ribosomal RNA processing, are quantitated and localized during oocyte maturation, fertilization, and early embryogenesis in the mouse. On the basis of Northern blot and nuclease protection experiments, it is estimated that there are about 5 x 10(4) U3 snRNA molecules in an ovulated oocyte and in a two-cell embryo. This number then increases roughly 50-fold to 2.7 x 10(6) molecules per embryo by the blastocyst stage. At all stages of development U3 snRNP antigens colocalize with nucleoli, as defined by differential interference contrast microscopy and an antibody to a nucleolar epitope. The synthesis and distribution of U3 snRNA and U3 snRNP follow a pattern independent from other major U snRNPs and snRNAs.

Animals↗