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Biomedical subjects

E C Carlà

Publications and source records attributed to E C Carlà.

5 recordsLinked to original sources

Lead nitrate and gadolinium chloride administration modify hepatocyte cell surfaces.

Modifications of hepatocyte cell surface were determined after single i.v. injection to rats of Pb(NO(3))(2) (known to induce liver hyperplasia followed by apoptosis) or GdCl(3) (known to induce proliferation of parenchymal cells and Kupffer cell depletion) or administration of GdCl(3) 24 h before Pb(NO(3))(2) injection (known to reduce hyperplasia and apoptosis induced in the parenchymal liver cells by the single Pb(NO(3))(2) injection). Rats were sacrificed at fixed times after the treatments (1, 3 and 5 days) and hepatocytes were isolated by enzymatic liver perfusion. In spite of the intracellular target of the heavy metals, signals leading to liver hyperplasia and apoptosis (with rates different for the different experimental conditions) were generated, which in turn were responsible for cell surface alteration. Increment or decrement of phosphatidylserine (PS) expression, asialoglycoprotein receptors (ASGPRs) and sugar residue expression on hepatocyte surfaces was measured in parallel with apoptosis and proliferation. When GdCl(3) was injected 24 h before Pb(NO(3))(2) injection, liver modifications were significantly reduced, thus suggesting that GdCl(3) could prevent and/or reduce liver damage.

Animals↗

Cell shape and plasma membrane alterations after static magnetic fields exposure.

The biological effects of static magnetic fields (MFs) with intensity of 6 mT were investigated in lymphocytes and U937 cells in the presence or absence of apoptosis-inducing drugs by transmission (TEM) and scanning (SEM) electron microscopy. Lectin cytochemistry of ConA-FITC conjugates was used to analyze plasma membrane structural modifications. Static MFs modified cell shape, plasma membrane and increased the level of intracellular [Ca++] which plays an antiapoptotic role in both cell types. Modifications induced by the exposure to static MFs were irrespective of the presence or absence of apoptotic drugs or the cell type. Abundant lamellar-shaped microvilli were observed upon 24 hrs of continuous exposure to static MFs in contrast to the normally rough surface of U937 cells having numerous short microvilli. Conversely, lymphocytes lost their round shape and became irregularly elongated; lamellar shaped microvilli were found when cells were simultaneously exposed to static MFs and apoptosis-inducing drugs. In our experiments, static MFs reduced the smoothness of the cell surface and partially impeded changes in distribution of cell surface glycans, both features being typical of apoptotic cells. Cell shape and plasma membrane structure modifications upon static MFs exposure were time-dependent. Lamellar microvilli were clearly observed before the distortion of cell shape, which was found at long times of exposure. MFs exposure promoted the rearrangement of F-actin filaments which, in turn, could be responsible for the cell surface modifications. Here we report data that support biological effects of static MFs on U937 cells and human lymphocytes. However, the involvement of these modifications in the onset of diseases needs to be further elucidated.

Adult↗

Synchronized onset of nuclear and cell surface modifications in U937 cells during apoptosis.

In this study we investigated the relationship between nuclear and cell surface modifications (i.e. blebbing, phosphatidylserine [PS] and sugar residues exposure) in a monocytic cell line, U937, during apoptosis induced by oxidative stress (1 mM H2O2) or inhibition of protein synthesis (10 microg/ml puromycin). Dying cells were simultaneously observed for nuclear modifications, presence of superficial blebs and plasma membrane alterations. Morphological analysis performed by conventional fluorescence microscopy, or by transmission and scanning electron microscopy showed that the courses of nuclear and membrane alterations occured concomitantly, but the phenotype was dependent on the stage of the apoptotic process and the type of apoptogenic inducer used. The progression of apoptosis in U937 cells beyond early stages resulted in the extensive formation of blebs which concomitantly lost some typical markers of apoptosis, such as PS and sugar residues. Therefore, the modality by which the nucleus condenses, or the amount and the pattern of distribution of PS on the cell surface were, for each cell line, strictly related to the apoptogenic inducer. The morphological data reported in the present paper should lead to a more precise quantification of apoptosis by improving the detection of apoptotic cells in vivo (i.e. in tissue, organs), which is a crucial point in the evaluation of efficiency of antiproliferative drugs, such as antiblastic or immunosuppressive compounds.

Apoptosis↗

Cell shape and organelle modification in apoptotic U937 cells.

U937 cells induced to apoptosis, progressively and dramatically modified their cell shape by intense blebbing formation, leading to the production of apoptotic bodies. The blebs evolved with time; milder forms of blebbing involving only a region or just the cortical part of the cytoplasm were observed within the first hour of incubation with puromycin; blebbing involving the whole cell body with very deep constrictions is the most frequent event observed during late times of incubation. The ultrastructural analysis of apoptotic cells revealed characteristic features of nuclear fragmentation (budding and cleavage mode) and cytoplasmatic modifications. The cytoplasm of blebs does not contain organelles, such as ribosomes or mitochondria. Scarce presence of endoplasmic reticulum can be observed at the site of bleb detachment. However, blebbing is a dispensable event as evaluated by using inhibitor of actin polymerization. In the present study, the progressive modifications of the nucleus, mitochondria, nuclear fragmentation, cytoplasmic blebs formation and production of apoptotic bodies in U937 monocytic cells induced to apoptosis by puromycin (an inhibitor of protein synthesis) were simultaneously analyzed.

Apoptosis↗

Relationship between cellular shape and receptor-mediated endocytosis: an ultrastructural and morphometric study in rat Kupffer cells.

The relationship between cellular shape (i.e., size, volume, presence of microvilli, pseudopodia, flat or round shape) and receptor-mediated endocytotic activities (i.e., binding and internalization) was investigated using intact liver as well as freshly isolated Kupffer cells and Kupffer cells in culture. The morphological features of Kupffer cells were reconstructed by three-dimensional analysis from in situ experiments and by densitometric analysis of cells in suspension and in culture. By morphometry at the ultrastructural level, different cellular shapes were compared with the respective capacities for binding and internalization of glycoproteins with terminal galactosyl residues. The number of asialoglycoprotein-gold particles bound to the cell surface or internalized into endosomes was calculated. Our data show that differences in cellular shape, mainly related to the reduction of projection and microvilli and to the roundness of cell surface, accompany modulation of galactose-specific receptors in rat Kupffer cells, thus supporting the hypothesis that cell morphology is affected by endocytic activities. In fact, the progressive reduction in microvilli projections and cellular roundness is paralleled by the progressive decrement of both binding and uptake capacity from in situ, freshly isolated and cultured Kupffer cells.

Animals↗